Luca FORTI - personale UniMoRe

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Luca FORTI

Professore Associato
Dipartimento di Scienze della Vita sede ex Chimica V.Campi 103

Pubblicazioni

2024 - Bioremediation of Basil Pesto Sauce-Manufactured Wastewater by the Microalgae Chlorella vulgaris Beij. and Scenedesmus sp [Articolo su rivista]
Scarponi, Paolina; Frongia, Francesca; Cramarossa, Maria Rita; Roncaglia, Fabrizio; Arru, Laura; Forti, Luca
abstract

Chlorella vulgaris and Scenedesmus sp. are commonly used in wastewater treatment due to their fast growth rates and ability to tolerate a range of environmental conditions. This study explored the cultivation of Chlorella vulgaris and Scenedesmus sp. using wastewater from the food industry, particularly from Italian basil pesto production tanks. The experiment involved different carbon dioxide concentrations and light conditions with a dilution rate of basil pesto wastewater at 1:2. Both microalgae strains were able to grow on pesto wastewater, and biomass characterization highlighted the influence of CO2 supply and light irradiation. The highest lipid storage was 79.3 ± 11.4 mg gdry biomass−1 and 75.5 ± 13.3 mg gdry biomass−1 for C. vulgaris and S. obliquus under red light (5% CO2 supply) and white light (0.04% CO2 supply), respectively. Protein storage was detected at 20.3 ± 1.0% and 24.8 ± 1.3% in C. vulgaris and S. obliquus biomasses under white light with a 5% CO2 and 0.04% CO2 supply, respectively. The removal of P, N, chemical oxygen demand, and biological oxygen demand resulted in 80–100%, 75–100%, 26–35%, and 0–20%, respectively.

2023 - Integrated Carbonate-Based CO2 Capture—Biofixation through Cyanobacteria [Articolo su rivista]
Ughetti, A.; Roncaglia, F.; Anderlini, B.; D’Eusanio, V.; Russo, A. L.; Forti, L.
abstract

2023 - Integrated wastewater treatment and CO2 capture by microalgae-based system [Articolo su rivista]
Frongia, F; Cramarossa, Mr; Arru, Laura; Forti, Luca
abstract

2023 - Lichens as a Natural Source of Compounds Active on Microorganisms of Human Health Interest [Articolo su rivista]
Condò, Carla; Anfelli, Igor; Forti, Luca; Sabia, Carla; Messi, Patrizia; Iseppi, Ramona
abstract

The antimicrobial properties of two lichen extracts (LC1 and LC2 solutions extracted in acetone and cyclohexane, respectively) were investigated against both Gram-positive and Gram-negative microorganisms through the agar well diffusion assay. Results displayed that both samples were similarly effective against all the indicator strains. The antimicrobial activity was maintained up to 30 days against Candida albicans ATCC 10231 with an inhibition zone of 38 mm and 37 mm for the LC1 and LC2 solutions extracted, respectively. In order to separate the single chemical components and to associate them with the biological activity, the two extracts were subjected to an activity-guided fractionation followed by a liquid chromatography mass spectrometry (LC–MS) Ion Trap 6310A for the chemical characterization. Chromatogram analysis of each sample that maintained an antimicrobial activity revealed the presence of a significant peak, at a retention time (tR) of 10.8 min, corresponding to a scabrosin derivative that could likely be associated with the antimicrobial activity. Results obtained in the present investigation, especially against the opportunistic pathogen C. albicans, are encouraging and could represent a preliminary step to a future solution toward a microorganism responsible for fungal infections, mainly occurring in immunocompromised patients and recently caused by drug-resistant strains.

2023 - New Perspectives on Monitoring Psychophysiological Wellbeing Effects after Inhaling Essential Oils — a Pilot Study [Relazione in Atti di Convegno]
Luca, Bonisoli; Marsili, Vittoria; Forti, L; Richeldi, D; Zamfirescu Richeldi, F; Arru, L
abstract

2023 - Unlocking Urban Insights: A Case Study on Impact of Urban Vegetation on Volatile Organic Compounds (VOCs) Variability Across Different Areas of Reggio Emilia, Italy [Articolo su rivista]
Marsili, Vittoria; Forti, Luca; Arru, Laura
abstract

2022 - Is there a role for sound in plants? [Articolo su rivista]
Del Stabile, F; Marsili, V; Forti, L; Arru, L
abstract

2022 - Microalgae potential in the capture of carbon dioxide emission. [Abstract in Atti di Convegno]
Frongia, F; Arru, L; Forti, L; Cramarossa, Mr
abstract

2022 - NEW SECONDARY METABOLITES IN THE AMPHINOMID FIREWORM HERMODICE CARUNCULATA [Poster]
Mucci, A.; Forti, L.; Simonini, R.; Ferrari, V.; Prevedelli, D.; Righi, S.
abstract

Eight betaine-derived novel compounds were found in extracts of the Mediterranean stinging fireworm Hermodice carunculata. The identification of their structures relies on 1D and 2D NMR (Fig. 1-3) and HPLC-ESI/HRMS spectra. Two types of terminal ammonium portions A and B and a series of different alkyl chains were identified (Fig. 4a,b). Their matching provides the structures of uncharacterized secondary metabolites, named carunculines, and their related isomers. These molecules differ from already known trimethylammonium inflammatory compounds (i.e. complanines) isolated from another amphinomid species, for the structures of the terminal ammonium groups (Fig. 4c) [1]. Carunculine anatomical distribution within H. carunculata was assessed by screening through HPLC-ESI/HRMS (Fig. 5, Table 1): their occurrence was revealed in all the body parts analyzed, both involved in predator-prey interactions [2], and mainly in the digestive apparatus. The results achieved reveal an array of different novel compounds from a chemically unknown species, improving knowledge on Marine Animal Products with chemical and biological potential for bioprospection [3]. Overall, these data reinforce the necessity of studying poorly-investigated taxa to expand knowledge on animal venom biology, their mechanisms of action and exploitation as promising source of drug molecules

2022 - NEW SECONDARY METABOLITES IN THE AMPHINOMID FIREWORM HERMODICE CARUNCULATA [Abstract in Atti di Convegno]
Mucci, A.; Forti, L.; Simonini, R.; Ferrari, V.; Prevedelli, D.; Righi, S.
abstract

Eight betaine-derived novel compounds were found in extracts of the Mediterranean stinging fireworm Hermodice carunculata. The identification of their structures relies on 1D and 2D NMR and HPLC-ESI/HRMS spectra (Fig. 1). Two types of terminal amine portions and a series of different alkyl chains were identified. Their matching provides the structures of uncharacterized secondary metabolites, named “carunculines”, and their related isomers. These molecules differ from already known trimethylammonium inflammatory compounds (i.e. “complanines”) isolated from another amphinomid species, for the structures of the terminal ammonium groups [1]. Carunculine anatomical distribution within H. carunculata was assessed by screening through HPLC-ESI/HRMS: their occurrence was revealed in all the body parts analyzed, both involved in predator-prey interactions [2], and mainly in the digestive apparatus. The results achieved reveal an array of different novel compounds from a chemically unknown species, improving knowledge on Marine Animal Products with chemical and biological potential for bioprospection. Overall, these data reinforce the necessity of studying poorly-investigated taxa to expand knowledge on animal venom biology, their mechanisms of action and exploitation as promising source of drug molecules.

2022 - Novel Natural Compounds and Their Anatomical Distribution in the Stinging Fireworm Hermodice carunculata (Annelida) [Articolo su rivista]
Righi, Sara; Forti, Luca; Simonini, Roberto; Ferrari, Valentina; Prevedelli, Daniela; Mucci, Adele
abstract

Increasing evidence in the field of bioprospection fosters the necessity of studying poorly investigated poisonous marine invertebrates to expand knowledge on animal venom biology. Among marine annelids, amphinomid fireworms are notorious for their bearded trunk equipped with a powerful stinging capacity. Here, a methodological workflow based on analytical chemistry techniques (compound isolation followed by mass spectrometry and spectroscopy analyses) was applied to gain new insights, leading to the identification and structural elucidation of an array of natural products from Mediterranean specimens of Hermodice carunculata. Eight betaine-derived unprecedented compounds, named "carunculines", were detected, bearing two terminal ammonium groups tri-and disubstituted at the Cα (A, B) and a series of different alkyl chains (I-VIII). The mixture of chemicals was found in all the body parts of H. carunculata, supporting a mechanism of action triggered by their vehiculation inside the dorsal chaetae, and subsequent injection when chaetae break off on contact. Preliminary investigations to understand adaptive features were also performed, showing a trend in carunculine abundance that fits into the evolutionary history of these worms. These findings shed light on the chemical ecology of amphinomids, giving reasons for the success of H. carunculata in benthic environments and providing promising novel metabolites for biotechnological implications.

2022 - Special Issue: "Cyanobacteria, Algae, and Plants; from Biology to Biotechnology" [Altro]
Forti, Luca; Arru, Laura; Bondi, Moreno
abstract

Photosynthetic organisms are milestones in the history of life: in fact, that they made it possible to shape Earth's atmosphere as we know it today and they are at the basis of almost all food chains, therefore, in a certain sense, of life on Earth. From a human point of view, plants have provided material to build shelter and procure medicine, food, and, of course, oxygen. Microalgae and cyanobacteria provide us almost half the oxygen we breathe and absorb a quarter of the CO2 produced by fossil fuels, and cyanobacteria are responsible for first introducing oxygen into the Earth’s anoxygenic atmosphere more than three billion years ago. Algae have often been associated with plants and classified accordingly, as they share some peculiar traits, and blue-green algae, or cyanobacteria, have been considered close to microalgae, since they derive energy from sunlight, like algae and plants through photosynthesis. However, even if they possess the same chlorophyll pigment and produce carbon dioxide, they do not have a nuclear membrane and, therefore, they are prokaryotes. Cyanobacteria, microalgae, and plants are beneficial and promising organisms for the sustainable production of food, feed, materials, chemicals, and fuels. To reach sustainability, considerable attention must be given to both strains and cultivars and available and new tools. From biology to biotechnology, research today should aim at eradicating hunger and illness in the world and at building a greener future. This Special Issue of Plants is focused on the most up-to-date research on these topics. In this Special Issue, we would like to present original research articles and reviews related but not limited to: - knowledge of and biotechnological applications for plant production, including specific aspects of sustainable agriculture and potential benefits to the environment and various other dimensions of human life - bio-sequestration of CO2 - remediation of polluted waters/soils - microalgal and cyanobacterial biomass and applications

2022 - Upgrading of Biobased Glycerol to Glycerol Carbonate as a Tool to Reduce the CO2 Emissions of the Biodiesel Fuel Life Cycle [Articolo su rivista]
Anderlini, Biagio; Ughetti, Alberto; Cristoni, Emma; Forti, Luca; Rigamonti, Luca; Roncaglia, Fabrizio
abstract

With regards to oil-based diesel fuel, the adoption of bio-derived diesel fuel was estimated to reduce CO2 emissions by approximately 75%, considering the whole life cycle. In this paper, we present a novel continuous-flow process able to transfer an equimolar amount of CO2 (through urea) to glycerol, producing glycerol carbonate. This represents a convenient tool, able to both improve the efficiency of the biodiesel production through the conversion of waste streams into added-value chemicals and to beneficially contribute to the whole carbon cycle. By means of a Design of Experiments approach, the influence of key operating variables on the product yield was studied and statistically modeled.

2021 - A tuning point in plant acoustics investigation [Articolo su rivista]
Allievi, Sara; Arru, Laura; Forti, Luca
abstract

In a very recent book called Sensory Biology of Plants, published by renowned publisher Springer Nature, the authors stated that the scientific literature gathered so far regarding knowledge around the field of Plant Acoustics allows us to divert the focus from the question "whether plants perceive sound" toward the questions "how and why they are doing it" Some phenomena are well known: roots perceive the sound of flowing water and display a sound-mediated growth toward the water source, while the buzz pollination process allows plants to minimize the pollen lost and maximize which is collected by true pollinators. But plants are far more perceptive and responsive to their environment than we generally consider them to be, and they are communicating far more information than we realize if we only took all their signals (VOCs, sound, exudates, etc.) into a greater picture. Could Volatile Organic Compounds (VOCs) be involved in mediating more responses than we imagine? VOC synthesis and release is known to be elicited also by electrical signals caused by mechanical stimuli, touching and wounding being among these, serving as info-chemicals in the communication between plants ("eavesdropping"), and within the organs of the same plant, in order for it to get synchronized with its surroundings. This paper is an overview of the discoveries around plant perception with a focus on the link between mechanical stimuli, as sound vibrations are, and changes in plant physiology leading to VOC emission.

2021 - An expedient catalytic process to obtain solketal from biobased glycerol [Articolo su rivista]
Roncaglia, F.; Forti, L.; D'Anna, S.; Maletti, L.
abstract

Developing simple and effective chemistry able to convert industrial waste streams into valuable chemicals is a primary contributor to sustainable development. Working in the context of biodiesel production, we found that plain bisulfate on silica (SSANa, 3.0 mmol/g) proved to be an optimal catalyst to convert glycerol into solketal. With the assistance of a proper anhydrification technique, isolated yields of 96% were achieved working in mild conditions, on 100 g scale.

2021 - Biological effect of different spinach extracts in comparison with the individual components of the phytocomplex [Articolo su rivista]
Arru, L.; Mussi, F.; Forti, L.; Buschini, A.
abstract

The Mediterranean-style diet is rich in fruit and vegetables and has a great impact on the prevention of major chronic diseases, such as cardiovascular diseases and cancer. In this work we investigated the ability of spinach extracts obtained by different extraction methods and of the single main components of the phytocomplex, alone or mixed, to modulate proliferation, antioxidant defense, and genotoxicity of HT29 human colorectal cells. Spinach extracts show dose-dependent activity, increasing the level of intracellular endogenous reactive oxygen species (ROS) when tested at higher doses. In the presence of oxidative stress, the activity is related to the oxidizing agent involved (H2O2 or menadione) and by the extraction method. The single components of the phytocomplex, alone or mixed, do not alter the intracellular endogenous level of ROS but again, in the presence of an oxidative insult, the modulation of antioxidant defense depends on the oxidizing agent used. The application of the phytocomplex extracts seem to be more effective than the application of the single phytocomplex components.

2021 - Le Creme delle Streghe [Monografia/Trattato scientifico]
Arru, L; Forti, L; Trentini, Ge
abstract

Il saggio (ri)scopre erbe e piante medicinali rintracciate nei manoscritti medievali come antichi rimedi tramandati per generazioni soprattutto dalle donne. Principali custodi di un incredibile patrimonio curativo, terapico e farmacologico, fondato su conoscenze naturali e senza basi scientifiche, furono avversate e perseguitate durante la famigerata "caccia alle streghe" proprio per questo. Il complesso lavoro di ricerca sui manoscritti ha permesso di scovare una quantità enorme di rimedi domestici, medici e veterinari di una modernità impressionante. Per esempio, si conoscevano già l'efficacia della bava di lumaca o delle lacrime di vite, un metodo per lo sbiancamento dei denti ed unguenti per le ustioni. Le ricette sono state tradotte, decifrate e studiate dagli autori, che rivedendo e modificando la composizione, gli ingredienti ed il procedimento botanico-farmaceutico, hanno dato nuova vita a rimedi naturali che conservano la forza di secoli di sapienza.

2021 - Microalgae potential in the capture of CO2 emission [Articolo su rivista]
Frongia, F.; Arru, L.; Cramarossa, M. R.; Forti, L.
abstract

In a perspective projected to reduce the atmospheric concentration of greenhouse gases, in which carbon dioxide is the master, the use of microalgae is an effective and decisive response. The review describes the bio circularity of the process of abatement of carbon dioxide through biofixation in algal biomass, highlighting the potential of its reuse in the production of high value-added products.

2021 - Synergy between mechanical injury and toxins triggers the urticating system of marine fireworms [Articolo su rivista]
Simonini, Roberto; Maggioni, Federica; Zanetti, Federica; Fai, Sergio; Forti, Luca; Prevedelli, Daniela; Righi, Sara
abstract

Marine fireworms (Annelida, Amphinomidae) are notorious for their stinging dorsal bristles (notochaetae). In the large-size species Hermodice carunculata, characterized by invasive potential and great predatory ability, notochaetae deter consumers and damage prey. The origin of fireworm defensive and offensive capacities, the mechanism of action of the stinging chaetae, and whether other chemical defenses occur are still uncertain. In the present study, we tested if the urticating system of fireworms is the result of a synergy between mechanical injury, due to notochaetae penetration, and the release of toxins vehiculated. To determine whether and where chemical or structural deterrents were present, different body parts of H. carunculata involved in inter-specific interactions were assayed (i.e. dorsal body wall, gills, dorsal and ventral parapodia). Feeding bioassays were performed to assess palatability against a …

2020 - A Chemical and Ecological Approach Sheds Light on the Urticating System of Marine Fireworms [Abstract in Atti di Convegno]
Righi, Sara; Savioli, Martina; Forti, Luca; Prevedelli, Daniela; Simonini, Roberto
abstract

Marine fireworms (Annelida, Amphinomidae) hold stinging dorsal bristles (chaetae) that cause injuries to divers and bathers. Hermodice carunculata is the most notorious species and it has recently attracted interest as a potentially invasive fireworm with few predators and uncharacterized defensive capacities. To date, the only acute inflammation inducer isolated from an amphinomid is “complanine”, a trimethylammonium compound. The main goal of this study was to promote an ecological understanding of H. carunculata defences through a multidisciplinary approach. The occurrence of complanine within tissues and its mode of delivery were assessed combining chromatographic steps and high resolution LC-MS/MS. The exact mass and retention time of complanine were detected in the extracts of the main ectodermal (dorsal body wall, gills, dorsal chaetae, ventral chaetae) and endodermal (gut, pharynx) tissues of H. carunculata. The role of complanine in trophic interactions was assessed offering the ectodermal tissues towards a predator (the fish Chromis viridis) and two relevant prey species (the anemones Anemonia viridis and Aiptasia diaphana). Only the dorsal chaetae were effective against predators and prey: they strongly deterred fishes and induced paralysis in the anemone tentacles. Dorsal chaetae treated with organic solvents lost their deterrence against fish predators and an inner hollow cavity suitable to vehicle toxins could be viewed by ESEM. These findings support a synergy between the mechanical injury of dorsal chaetae penetration and the release of complanine. This unique feature could support the success of fireworms in marine benthic environments and significantly improves knowledge on the chemical ecology of amphinomids.

2020 - Antimicrobial activity of spices essential oils and its effectiveness on mature biofilms of human pathogens [Articolo su rivista]
Condò, Carla; Anacarso, Immacolata; Sabia, Carla; Iseppi, Ramona; Anfelli, Igor; Forti, Luca; de Niederhäusern, Simona; Bondi, Moreno; Messi, Patrizia
abstract

The antibacterial activity of Pimpinella anisum L., Cinnamomum zeylanicum, Syzygium aromaticum, and Cuminum cyminum L. essential oils (EOs) against some common pathogenic microorganisms (Staphylococcus aureus ATCC 6538, Staphylococcus epidermidis ATCC 14990, Enterococcus faecalis ATCC 29212, Streptococcus pyogenes ATCC 1915, Escherichia coli ATCC 8739, Pseudomonas aeruginosa ATCC 27853, Aeromonas hydrophila ATCC 7966, Proteus mirabilis ATCC 10005, Klebsiella pneumoniae ATCC 13883, and Candida albicans ATCC 10231) and their biofilms was studied. The EOs inhibitory effects were evaluated by both Agar Well Diffusion assay and Minimum Inhibitory Concentration (MIC) determination. The most active EOs, cinnamon and cloves, were also tested on 18, 24, 48, 72 hours mature biofilms. Cinnamon and cloves exhibited the best results showing a significant activity against all the tested bacteria. Concerning biofilm, results suggest that Cinnamomum zeylanicum oil may be a useful approach to impair the biofilm produced by the tested Gram-negative bacteria.

2020 - Non-conventional yeasts as sources of ene-reductases for the bioreduction of chalcones [Articolo su rivista]
Filippucci, S.; Tasselli, G.; Kenza Labbani, F. -Z.; Turchetti, B.; Rita Cramarossa, M.; Buzzini, P.; Forti, L.
abstract

Thirteen Non-Conventional Yeasts (NCYs) have been investigated for their ability to reduce activated C=C bonds of chalcones to obtain the corresponding dihydrochalcones. A possible correlation between bioreducing capacity of the NCYs and the substrate structure was estimated. Generally, whole-cells of the NCYs were able to hydrogenate the C=C double bond occurring in (E)-1,3-diphenylprop-2-en-1-one, while worthy bioconversion yields were obtained when the substrate exhibited the presence of a deactivating electron-withdrawing Cl substituent on the B-ring. On the contrary, no conversion was generally found, with a few exceptions, in the presence of an activating electron-donating substituent OH. The bioreduction aptitude of the NCYs was apparently correlated to the logP value: Compounds characterized by a higher logP exhibited a superior aptitude to be reduced by the NCYs than compounds with a lower logP value.

2020 - Sound perception and its effects in plants and algae [Articolo su rivista]
Frongia, F.; Forti, L.; Arru, L.
abstract

Life evolved in an acoustic world. Sound is perceived in different ways by the species that inhabit the Planet. Among organisms, also some algal species seem to respond to sound stimuli with increased cell growth and productivity. The purpose of this Short Communication is to provide an overview of the current literature about various organisms and sound, with particular attention to algal organisms, which, when subjected to sound applications, can change their metabolism accordingly.

2019 - Ecotoxicity of hallachrome, an unusual 1-2 anthraquinone excreted by the infaunal polychaete Halla parthenopeia: evidence for a chemical defence? [Articolo su rivista]
Simonini, R; Iori, D; Forti, L; Righi, S; Prevedelli, D
abstract

Polychaetes play a prominent role in marine systems, but little is known about their secondary metabolites compared with other benthic taxa. In the present study, we investigated the toxicity of hallachrome, an unusual 1-2 anthraquinone identified from the skin of some polychaetes, including the Mediterranean infaunal species Halla parthenopeia. Under stress conditions, this worm releases a harmful purple mucus, whose noxious compounds were still unknown. We hypothesized that hallachrome also occurs in the purple mucus, giving rise to its color and toxicity. Soon after the production of the purple exudate, H. parthenopeia also secretes a harmless, transparent mucus, which pushes away the toxic one, suggesting protective functions for the worm itself. LC-MS and H-1-NMR analyses confirmed the presence of the pigment hallachrome in the purple mucus. The average concentration of the pigment in the purple mucus was about 310 mg L-1. Ecotoxicological bioassays on representative species of bacteria, protozoans, rotifers, crustaceans (Artemia franciscana) and polychaetes (Dinophilus gyrociliatus) revealed its severe toxic effects: LC50/EC(50)values ranged from 0.11-5.67 mg L-1. Hallachrome showed higher toxicity for A. franciscana than other naturally occurring anthraquinones. Tests on encapsulated embryos of D. gyrociliatus evidenced the ability of a mucus layer to limit hallachrome diffusion, confirming the protective role of the transparent mucus. Given the information available on polychaetes anti-predator strategies, hallachrome cannot be considered a consumer deterrent. However its toxicity and wide range of activity suggest chemical defensive functions against potential competitors, parasites and/or pathogens.

2019 - Oxygen Availability during Growth Modulates the Phytochemical Profile and the Chemo-Protective Properties of Spinach Juice [Articolo su rivista]
Milano, Francesco; Mussi, F; Fornaciari, S; Altunoz, M; Forti, L; Arru, L; Buschini, A
abstract

Fruits and vegetables are a good source of potentially biologically active compounds. Their regular consumption in the human diet can help reduce the risk of developing chronic diseases such as cardiovascular diseases and cancer. Plants produce additional chemical substances when subject to abiotic stress or infected by microorganisms. The phytochemical profile of spinach leaves (Spinacia oleracea L.), which is a vegetable with widely recognized health-promoting activity, has been affected by applying root hypoxic and re-oxygenation stress during plant growth. Leaf juice at different sampling times has been subject to liquid chromatography mass spectrometry (LCMSn) analysis and tested on the human colorectal adenocarcinoma cell line HT29 by using the Comet assay. The cells were previously treated with H2O2 to simulate the presence of an oxidative stress (as in colon cancer condition) and the leaf juice application resulted in a significant antioxidant and protective in vitro effect. The duration of the hypoxic/re-oxygenation stress imposed on the plant reflects the antioxidant leaf juice content. After hypoxic stress (24 hours) and reoxygenation (2 hours), we show a decrease (50%) of the relative abundance of the principal identified antioxidant molecules but a higher antioxidant activity of the spinach juice on HT29 cells (20%). Data shows a complex relation between plant growing conditions and the modulation of secondary metabolites content in leaf juice that results in different chemo-protective activities in colon cancer cells.

2019 - Synthesis and anticancer activity of CDDO and CDDO-me, two derivatives of natural triterpenoids [Articolo su rivista]
Borella, R.; Forti, L.; Gibellini, L.; De Gaetano, A.; De Biasi, S.; Nasi, M.; Cossarizza, A.; Pinti, M.
abstract

Triterpenoids are natural compounds synthesized by plants through cyclization of squalene, known for their weak anti-inflammatory activity. 2-cyano-3,12-dioxooleana-1,9(11)-dien-28-oic acid (CDDO), and its C28 modified derivative, methyl-ester (CDDO-Me, also known as bardoxolone methyl), are two synthetic derivatives of oleanolic acid, synthesized more than 20 years ago, in an attempt to enhance the anti-inflammatory behavior of the natural compound. These molecules have been extensively investigated for their strong ability to exert antiproliferative, antiangiogenic, and antimetastatic activities, and to induce apoptosis and differentiation in cancer cells. Here, we discuss the chemical properties of natural triterpenoids, the pathways of synthesis and the biological effects of CDDO and its derivative CDDO-Me. At nanomolar doses, CDDO and CDDO-Me have been shown to protect cells and tissues from oxidative stress by increasing the transcriptional activity of the nuclear factor (erythroid-derived 2)-like 2 (Nrf2). At doses higher than 100 nM, CDDO and CDDO-Me are able to modulate the differentiation of a variety of cell types, both tumor cell lines or primary culture cell, while at micromolar doses these compounds exert an anticancer effect in multiple manners; by inducing extrinsic or intrinsic apoptotic pathways, or autophagic cell death, by inhibiting telomerase activity, by disrupting mitochondrial functions through Lon protease inhibition, and by blocking the deubiquitylating enzyme USP7. CDDO-Me demonstrated its efficacy as anticancer drugs in different mouse models, and versus several types of cancer. Several clinical trials have been started in humans for evaluating CDDO-Me efficacy as anticancer and anti-inflammatory drug; despite promising results, significant increase in heart failure events represented an obstacle for the clinical use of CDDO-Me.

2018 - Nonconventional Yeast-Promoted Biotransformation for the Production of Flavor Compounds [Capitolo/Saggio]
Forti, Luca; Cramarossa, Maria R.; Filippucci, Sara; Tasselli, Giorgia; Turchetti, Benedetta; Buzzini, Pietro
abstract

The rising consumer requests for natural flavors and fragrances has generated a great interest in the aroma industry to seek new methods to obtain fragrance and flavor compounds naturally. An alternative and attractive route for these compounds is based on biotransformations. In this context, non-conventional yeasts (NCYs) have attracted increasing interest due to their biochemical characteristics and potential applications, being able to produce aroma compounds from a variety of carbon sources, including sugars, alkanes, plant oils, starch hydrolysates, ethanol, and glycerol. Apart from classical fermentation processes (de novo synthesis), bioconversion of appropriate precursor compounds are also being developed to produce food aromas. An overview on the potential of NCYs whole cell for producing food flavors by biotransformation is illustrated in this chapter by a discussion of the production of different class of compounds, namely Aldehydes, Ketones and related compounds, Alcohols, Lactones, Terpenes and Terpenoids, Alkenes, and Phenols.

2017 - Combined Effects of LED Lights and chicken manure on Neochloris oleoabundans Growth [Articolo su rivista]
ALTUNOZ HATIPOGLU, Meltem; Pirrotta, Onofrio; Forti, Luca; Allesina, Giulio; Pedrazzi, Simone; Obali, Olcay; Tartarini, Paolo; Arru, Laura
abstract

In this study a photobioreactor prototype is presented for the culture growth of microalgae model organism Neochloris oleoabundans by using chicken manure waste as feedstock along with the optimum combination of led light wavelengths and light intensity. Particularly interesting results are observed on the strains fed by chicken manure medium under the proper combination of red and blue LED light illumination, the microalgal growth resulted comparable with the strains fed by the costly commercial microalgal growth medium (BG 11 medium). Cell concentration, optical density, growth rate, cell size, total lipid and photosynthetic pigment content have been monitored during a time-course experiment. The data suggest that there are difficulties due to white light diffusion into the dark chicken medium, which leads to a generally lower intensity scattered along all wavelengths; blue or combined red and blue lights resulted in a higher irradiation density, affecting microalgae cell growth.

2017 - PROVE DI ADATTAMENTO DELLA CANAPA (CANNABIS SATIVA L.) NELL'APPENNINO DELL’EMILIA CENTRALE [Abstract in Atti di Convegno]
Ronga, Domenico; Caradonia, Federica; Setti, Leonardo; Hagassou, Djangsou; Arru, Laura; Forti, Luca; Endrighi, Emiro; Bazzani, Grassi G.; Francia, Enrico
abstract

La canapa (Cannabis sativa L.) è una pianta utilizzata per molteplici usi. In Europa, tra il XVI e il XVII secolo, era una delle colture principali impiegate per la produzione di fibre tessili. Già a partire da fine ‘800 la mancata industrializzazione, il proibizionismo e la disponibilità di nuove fibre più competitive hanno portato al declino della canapicoltura. Negli ultimi anni però, caratteristiche positive quali il basso impatto ambientale, la capacità di ridurre il consumo dei suoli, le basse esigenze colturali, nonché il suo possibile uso in diversi settori (alimentare, erboristico, della bioedilizia, ecc.) hanno portato a riconsiderare la sua coltivazione. La canapa, infatti, è una coltura dinamica che si adatta a diverse condizioni agro-ambientali e che non ha grandi esigenze idriche, nutritive e fitosanitarie. Pertanto, potrebbe essere coltivata con buone rese anche in ambienti marginali come ad esempio quelli dell’Appennino emiliano. Lo scopo di questo studio è stato quello di valutare l’adattamento di diverse cultivar moderne di canapa in un’azienda sperimentale biologica, situata nell’Appennino dell’Emilia centrale, per la produzione di infiorescenze da destinare all’estrazione di oli essenziali. La prova è stata effettuata nel 2016 presso l’azienda dell’Istituto “Lazzaro Spallanzani”, Montombraro (Modena). Tre diverse cultivar, Uso 31, Futura 75 e Fedora 17, sono state seminate nella seconda decade di giugno secondo uno schema sperimentale a blocchi completamente randomizzati. Durante la prova non sono stati utilizzati fertilizzanti e prodotti fitosanitari ed è stata effettuata solo un’irrigazione di soccorso all’emergenza. Sono stati poi rilevati i seguenti parametri: emergenza piante, altezza della pianta, diametro colletto, biomassa aerea fresca (di infiorescenze, fusti e foglie), indice relativo al contenuto in clorofilla mediante SPAD-502. Dai rilievi effettuati è emerso che tutte e 3 le cultivar si sono adattate bene alle condizioni di coltivazione ma la cultivar Futura 75 è quella che si è adattata meglio all’ambiente di coltivazione ed ha mostrato una produzione di biomassa fresca di infiorescenze superiore alle altre 2 cultivar. Sono in corso prove di estrazione degli oli essenziali dai campioni di biomassa raccolti mediante distillatore. La possibilità di poter coltivare la canapa potrebbe da un lato migliorare il reddito dell’agricoltore e dall’altro avere effetti positivi sull’agro-ecosistema permettendo così il rilancio e la valorizzazione delle aree dell’Appennino emiliano.

2016 - Integrated Analysis of the Wood Oil from Xanthocyparis vietnamensis Farjon & Hiep. By Chromatographic and Spectroscopic Techniques [Articolo su rivista]
Bazzali, O.; Thai, T. H.; Hoi, T. M.; Khang, N. S.; Hien, N. T.; Casanova, J.; Bighelli, A.; Tomi, F.; Forti, L.
abstract

In order to get better knowledge about the volatiles produced by Xanthocyparis vietnatnensis, a species recently discovered in Vietnam, its wood oil has been analyzed by a combination of chromatographic (GC, CC) and spectroscopic (GC-MS, 13C-NMR) techniques. Forty components that accounted for 87.9% of the oil composition have been identified. The composition is dominated by nootkatene (20.7%), 11,12,13-tri-nor-eremophil-1(10)-en-7-one (17.2%), γ-eudesmol (5.1%), nootkatone (4.7%), valencene (3.5%) and 13-nor-eremophil-l(10)-en-11-one (2.6%). The structure of two new compounds-10-epi-nor-γ-eudesmen-11-one and 12-hydroxy-isodihydroagarofuran-has been elucidated, while 11,12,13-tri-nor-eremophil-1(10)-en-7-ol is reported as a natural product for the first time. The composition of X. vietnamensis wood oil varied drastically from those of leaf oils, dominated by hedycaryol (34.4%), phyllocladene (37.8%) or by pimara-6(14)-15-diene (19.4%).

2016 - Study of Holtermanniella wattica, Leucosporidium creatinivorum, Naganishia adeliensis, Solicoccozyma aeria, and Solicoccozyma terricola for their lipogenic aptitude from different carbon sources [Articolo su rivista]
Filippucci, Sara; Tasselli, Giorgia; Scardua, Alessandro; Di Mauro, Simone; Cramarossa, Maria Rita; Perini, Davide; Turchetti, Benedetta; Onofri, Andrea; Forti, Luca; Buzzini, Pietro
abstract

Background The ability of some microorganisms to accumulate lipids is well known; however, only recently the number of studies on microbial lipid biosynthesis for obtaining oleochemical products, namely biofuels and some building blocks for chemistry, is rapidly and spectacularly increased. Since 1990s, some oleaginous yeasts were studied for their ability to accumulate lipids up to 60–70% of their dry weight. Due to the vast array of engineering techniques currently available, the recombinant DNA technology was the main approach followed so far for obtaining lipid-overproducing yeasts, mainly belonging to the Yarrowia lipolytica. However, an alternative approach can be offered by worldwide diversity as source of novel oleaginous yeasts. Lipogenic aptitude of a number of yeast strains has been reviewed, but many of these studies utilized a limited number of species and/or different culture conditions that make impossible the comparison of different results. Accordingly, the lipogenic aptitude inside the yeast world is still far from being fully explored, and finding new oleaginous yeast species can acquire a strategic importance. Results Holtermanniella wattica, Leucosporidium creatinivorum, Naganishia adeliensis, Solicoccozyma aeria, and Solicoccozyma terricola strains were selected as a result of a large-scale screening on 706 yeasts (both Ascomycota and Basidiomycota). Lipid yields and fatty acid profiles of selected strains were evaluated at 20 and 25 °C on glucose, and on glycerol, xylose, galactose, sucrose, maltose, and cellobiose. A variable fatty acid profile was observed in dependence of both temperature and different carbon sources. On the whole, L. creatinivorum exhibited the highest performances: total lipid yield (YL) >7 g/l on glucose and glycerol, % of intracellular lipids on cell biomass (YL/DW) >70% at 20 °C on glucose, lipid coefficient (YL/Glu) around 20% on glucose, and daily productivity (YL/d) on glucose and sucrose >1.6 g/(l*d). Conclusions This study provides some meaningful information about the lipogenic ability of some yeast species. Variable lipid yields and fatty acid profiles were observed in dependence of both temperature and different carbon sources. L. creatinivorum exhibited the highest lipogenic performances.

2016 - The investigation of resveratrol and analogs as potential inducers of fetal hemoglobin [Articolo su rivista]
Theodorou, Andria; Phylactides, Marios; Forti, Luca; Cramarossa, Maria Rita; Spyrou, Pantelis; Gambari, Roberto; Thein, Swee Lay; Kleanthous, Marina
abstract

Beta-thalassemia, is a hemoglobinopathy characterized by reduced beta-globin chain synthesis, leading to imbalanced globin chain production, ineffective erythropoiesis and anemia. Increasing gamma-globin gene expression is a promising therapeutic approach as it reduces this imbalance by combining with the excess alpha globin chains and producing fetal hemoglobin (HbF). Furthermore, increased iron absorption and repeated blood transfusions lead to iron overload and tissue damage secondary to reactive oxygen species. Compounds exhibiting both antioxidant and HbF inducing activities are, therefore, highly desirable therapeutic agents. Resveratrol, a natural phytoalexin, combines these two activities but is also cytotoxic. Nine hydroxystilbenic resveratrol derivatives were synthesized in an attempt to identify compounds that retain the HbF-inducing and antioxidant activities of resveratrol but exhibit reduced cytotoxicity. Three derivatives (P1, P4 and P11) exhibited similar hemoglobin-inducing properties to resveratrol in K562 cells, however, only P11 showed reduced cytotoxicity. All three derivatives demonstrated variable HbF-inducing activity in primary erythroid progenitor cells from healthy donors. Resveratrol and P11 increased HbF induction significantly, with P11 having the highest activity. Additionally, P4 significantly increased progenitor numbers. A combinatorial treatment in K562 cells using resveratrol and decitabine resulted in a statistically significant increase in hemoglobin-inducing activity only above the level shown by resveratrol alone.

2016 - Valorization of glass wastes as support for lipase immobilization [Articolo su rivista]
Barbieri, Luisa; Bursi, Elena; Cramarossa, Maria Rita; Ferroni, Laura; Forti, Luca; Lancellotti, Isabella; Ponzoni, Chiara; Vassura, Ivano
abstract

This work focuses on treatment of lead polluted waste glass from urban collection (Pb≈5% wt) for its valorization as support in the immobilization of lipase, because soda-lime glasses are used in enzymatic catalysis. First of all, an extraction process of surface Pb was performed. The process is based on nitrilotriacetic acid (NTA) chelating agent. The operating conditions are: T=80°C, t=1h, pH 10, solid/liquid weight ratio=1/10, reagent concentration=0.1 M. Leaching tests at controlled pH performed on treated glass confirmed the typical inert nature of the glass and the effectiveness of the NTA treatment. Afterwards lead polluted waste glasses, both untreated and treated with NTA, together with a commercial reference glass, were used as physical supports for the adsorption of Rhizopus oryzae lipase, in order to investigate their immobilization capability after specific washing/activation treatments to make functional their surface. The biocatalytic activity of the immobilized enzyme on the glasses was tested through the hydrolysis of para-nitrophenyl acetate to para-nitrophenol, quantifying the conversion percentage after 60 min of reaction by means of UV analyses at 410 nm. From the first results, lipase immobilization on waste glasses appears to be encouraging. In particular, lipase immobilized on lead polluted glass from urban collection showed an activity comparable to that of reference glass.

2015 - Analisi preliminari sulla tossicità di essudati di Arenicola marina (Linnaeus, 1758) (Annelida: Arenicolidae) [Articolo su rivista]
Iori, D.; Mirra, M.; Forti, Luca; Prevedelli, Daniela; Simonini, Roberto
abstract

The effects of the exudate (crude and its organic extracts) obtained by Arenicola marina (Annelida: Arenicolidae) were evaluated through acute toxicity bioassays on the annelid Dinophilus gyrociliatus. Preliminary chemical analyses were also performed. The exudate and its n-buthanol extract were toxic (LC50 24h=12%, c.i.=9.7-14.7% and 31.7 mg l-1, c.i.=27.9-36 mg l-1, respectively). The results support the hypothesis that the exudate could be involved in the chemical defence of A. marina.

2015 - Assessment of antioxidant and antiproliferative properties of spinach plants grown under low oxygen availability [Articolo su rivista]
Fornaciari, Silvia; Milano, F.; Mussi, Francesca; Pinto Sanchez, L.; Forti, Luca; Buschini, A.; Arru, Laura
abstract

Background In the human diet, consumption of fresh fruits and vegetables is important in maintaining a good health status and in preventing chronic diseases. It is known that plant derived food is a powerful source of chemopreventive molecules i.e. antioxidants, and spinach (Spinacia oleracea L., Chenopodiaceae) possesses a wide range of metabolites with such biological activity. Plant stress response could lead to the production of metabolites with high value for human health and this could be a tool to enhance the production of molecules with antioxidant activity in plants. Results Data reported in this paper confirm the antioxidant properties of spinach plants, and show a strong antiproliferative activity of leaf extract on HT-29 human cell line. Besides, the hypoxic stress seems to affect the pool of antioxidant molecules present in spinach leaves, as verified by means of HPLC-MS/MS analysis and the aluminium chloride and ABTS assays. Conclusion Our findings - represents a hint for an improvement in the biological and pharmacological properties of spinach plants, being the growth condition a strategy to modulate the spinach phytocomplex profile.

2015 - Chemical composition and biological activities of fragrant mexican copal (Bursera spp.) [Articolo su rivista]
Gigliarelli, G.; Becerra, J. X.; Curini, M.; Marcotullio, M. C.; Forti, L.
abstract

Copal is the Spanish word used to describe aromatic resins from several genera of plants. Mexican copal derives from several Bursera spp., Protium copal, some Pinus spp. (e.g., P. pseudostrobus) and a few Fabaceae spp. It has been used for centuries as incense for religious ceremonies, as a food preservative, and as a treatment for several illnesses. The aim of this review is to analyze the chemical composition and biological activity of commercial Mexican Bursera copal.

2015 - Chemo-enzymatic synthesis of new resveratrol-related dimers containing the benzo[b]furan framework and evaluation of their radical scavenger activities [Articolo su rivista]
Beneventi, Elisa; Conte, Silvia; Cramarossa, Maria Rita; Riva, Sergio; Forti, Luca
abstract

The chemo-enzymatic synthesis and the evaluation of radical scavenger performance of resveratrolrelated dimers with antioxidative power have been investigated. The dimeric compounds, containing the benzo[b]furan framework, were prepared via an oxidative dimerization catalyzed by a laccase from Trametes versicolor, followed by a treatment with the organic oxidant DDQ. This methodology can be useful for the synthesis of various 2,3-diarylbenzo[b]furans derivates, a class of compound that exhibits a wide range of biological activities.

2015 - Non-Conventional Yeasts Whole Cells as Efficient Biocatalysts for the Production of Flavors and Fragrances [Articolo su rivista]
Forti, Luca; Di Mauro, Simone; Cramarossa, Maria Rita; Filippucci, Sara; Turchetti, Benedetta; Buzzini, Pietro
abstract

The rising consumer requests for natural flavors and fragrances have generated great interest in the aroma industry to seek new methods to obtain fragrance and flavor compounds naturally. An alternative and attractive route for these compounds is based on bio-transformations. In this review, the application of biocatalysis by Non Conventional Yeasts (NCYs) whole cells for the production of flavor and fragrances is illustrated by a discussion of the production of different class of compounds, namely Aldehydes, Ketones and related compounds, Alcohols, Lactones, Terpenes and Terpenoids, Alkenes, and Phenols.

2014 - Corrigendum to "Biotransformation of electron-poor alkenes by yeasts: Asymmetric reduction of (4S)-(+)-carvone by yeast enoate reductases" [Enzyme Microb. Technol. 45 (2009) 463-468] [Articolo su rivista]
Goretti, M.; Ponzoni, C.; Caselli, E.; Marchegiani, E.; Cramarossa, M. R.; Turchetti, B.; Buzzini, P.; Forti, L.
abstract

2014 - Difese chimiche in anellidi marini: valutazione della tossicità dell’hallacromo prodotto da Halla parthenopeia (Polychaeta: Oenonidae) [Abstract in Atti di Convegno]
Simonini, Roberto; Iori, Dario; Forti, Luca; MASSAMBA N'SIALA, Gloria; Prevedelli, Daniela
abstract

Alcuni anellidi marini producono metaboliti secondari per difendersi chimicamente contro predatori, competitori e/o patogeni. L’hallacromo (C16H12O4) è un antrachinone presente nel muco violaceo prodotto dall'anellide marino Halla parthenopeia in risposta a stress. Abbiamo recentemente osservato che l’esposizione al muco violaceo è nociva o letale per organismi-test appartenenti a taxa molto diversi. In questo lavoro la tossicità dell’hallacromo (estratto dal muco, purificato tramite cromatografia e diluito in DMSO dopo caratterizzazione tramite LC-MS e NMR) è stata valutata tramite una batteria di saggi ecotossicologici. L’hallacromo è letale a basse concentrazioni per il polichete Dinophilus gyrociliatus (LC50 96 h= 0.11 mg l-1, NOEC=0.026 mg l-1) e presenta una elevata tossicità anche per gli altri organismi testati (Brachionus plicatilis: LC50 24 h= 0.16 mg l -1; Euplotes crassus: LC50 2h= 0.29 mg l-1; Vibrio fischeri: EC50 30’= 0.77 mg l-1; Artemia francescana: LC50 24h= 5.67-7.05 mg l-1). I risultati suggeriscono che l’hallacromo sia il responsabile dell’attività biologica del muco violaceo di H. parthenopeia. L'elevata tossicità e l'ampia gamma di organismi su cui esercita effetti suggerisce che l’hallacromo rappresenti una difesa chimica verso competitori e parassiti. Di norma gli studi sulle difese chimiche degli invertebrati marini si basano su test di palatabilità, che evidenziano soprattutto le difese antipredatorie. Integrare i test di palatabilità con saggi ecotossicologici potrebbe contribuire al miglioramento delle conoscenze sui differenti tipi di difese chimiche messe in atto dagli invertebrati marini.

2014 - Tossicitá del muco colorato di Halla parthenopeia (Delle Chiaje, 1828) (Annelida, Oenonidae) [Articolo su rivista]
Iori, Dario; Forti, Luca; Prevedelli, Daniela; N’Siala, G. Massamba; Simonini, Roberto
abstract

The toxic effects of the coloured mucus produced under stress conditions by Halla parthenopeia (Polychaeta: Oenonidae) was assessed by means of a battery of ecotoxicological tests that included bacteria, protozoan, rotifers, polychaetes and crustaceans. Coloured mucus was toxic at concentrations ranging from 70.000 to 300 mg l-1 (in terms of LC50 values) depending on the tested species. Results suggest that the coloured mucus should play a role in the chemical defence of H. parthenopeia.

2014 - Toxicity of the purple mucus of the polychaete Halla parthenopeia (Oenonidae) revealed by a battery of ecotoxicological bioassays [Articolo su rivista]
Iori, Dario; Forti, Luca; MASSAMBA N'SIALA, Gloria; Prevedelli, Daniela; Simonini, Roberto
abstract

Mucus secretions play a number of functions related to polychaete physiology and ecology. Under stress conditions, the polychaete Halla parthenopeia (Oenonidae) produces a purple mucus after mechanical stimulation, whose function is still unknown. Here, we assessed the toxicity of this purple mucus by means of both acute toxicity bioassays on the polychaete Dinophilus gyrociliatus and commercial ecotoxicological kits (Microtox®, Rotoxkit® and Artoxkit®). Palatability was also tested with the fish Oryzias melastigma. After emitting purple mucus, H. parthenopeia quickly moves away and starts releasing transparent mucus. Acute toxicity bioassays showed that the mucus was harmless (transparent), or lethal even when diluted about 1000 times (purple). Purple mucus was toxic at different concentrations, the LC50 ranging from 0.7-0.3 g l–1 for D. gyrociliatus to 76 g l–1 for Artemia franciscana (Artoxkit®). Freeze-dried brine shrimp coated with transparent or purple muci were both consumed by O. melastigma. We hypothesized that the purple mucus is involved in the chemical defence of H. parthenopeia against competitors and parasites, and that its colour and toxicity are due to hallachrome, a 1,2-anthraquinone found in the skin of H. parthenopeia.

2013 - Fungal laccases and enoate reductases as biocatalysts of fine chemical transformations [Relazione in Atti di Convegno]
A., Romagnolo; F., Spina; D., Carusetta; L., Nerva; Cramarossa, Maria Rita; F., Parmeggiani; Forti, Luca; E., Brenna; G. C., Varese
abstract

Laccase and enoate reductase are promising tools in bioconversion of several substrates of biotechnological interest. In this study, one fungus producing laccase and two fungi producing enoate reductase were used in the bioconversion of caffeic acid and a set of unsaturated compounds (α- methylcinnamaldehyde, α- methylnitrostyrene and cyclohexenone) showing interesting conversion products.

2013 - Production of Flavours and Fragrances via Bioreduction of (4R)-(-)-Carvone and (1R)-(-)-Myrtenal by Non-Conventional Yeast Whole-Cells [Articolo su rivista]
M., Goretti; B., Turchetti; Cramarossa, Maria Rita; Forti, Luca; P., Buzzini
abstract

As part of a program aiming at the selection of yeast strains which might be of interest as sources of natural flavours and fragrances, the bioreduction of (4R)-(−)-carvone and (1R)-(−)-myrtenal by whole-cells of non-conventional yeasts (NCYs) belonging to the genera Candida, Cryptococcus, Debaryomyces, Hanseniaspora, Kazachstania, Kluyveromyces, Lindnera, Nakaseomyces, Vanderwaltozyma and Wickerhamomyces was studied. Volatiles produced were sampled by means of headspace solid-phase microextraction (SPME) and the compounds were analysed and identified by gas chromatography–mass spectroscopy (GC-MS). Yields (expressed as % of biotransformation) varied in dependence of the strain. The reduction of both (4R)-(−)-carvone and (1R)-(−)-myrtenal were catalyzed by some ene-reductases (ERs) and/or carbonyl reductases (CRs), which determined the formation of (1R,4R)-dihydrocarvone and (1R)-myrtenol respectively, as main flavouring products. The potential of NCYs as novel whole-cell biocatalysts for selective biotransformation of electron-poor alkenes for producing flavours and fragrances of industrial interest is discussed.

2012 - Asymmetric reduction of (4S)-(+)-carvone catalyzed by enoate reductases (ERs) expressed by non-conventional yeast (NCY) whole-cells [Capitolo/Saggio]
P., Buzzini; M., Goretti; C., Ponzoni; E., Caselli; E., Branda; Cramarossa, Maria Rita; B., Turchetti; Forti, Luca
abstract

(4S)-(þ)-Carvone was totally reduced after 120 h by ER activity expressed by lyophilized whole cells of K. exigua DBVPG 7133 to (1S,4S)-dihydrocarvone (yield¼60%) and (1R,4S)-dihydrocarvone (40%). About 15% of (1S,4S)-dihydrocarvone was further reduced to (1S,2S,4S)-dihydrocarveol (7%) and (1S,2R,4S)-dihydrocarveol (8%). In close analogy with previous literature, the use of lyophilized yeast whole cells was generally preferred to growing and resting ones. Besides, because yeast cells contain only catalytic amount of NAD(P)H and cofactor regeneration must take place by means of metabolism of an electron donor, the simultaneous use of lyophilized yeast whole cells and glucose (the last acting as auxiliary substrate for cofactor-recycling systems) showed a significant increase of efficiency in reducing (4S)-(þ)-carvone.

2012 - Response surface methodology as optimization strategy for asymmetric bioreduction of (4S)-(+)-carvone by Cryptococcus gastricus [Articolo su rivista]
M., Goretti; E., Branda; B., Turchetti; Cramarossa, Maria Rita; A., Onofri; Forti, Luca; P., Buzzini
abstract

Response surface methodology was applied in optimizing the asymmetric bioreduction of (4S)-(+)-carvoneto dihydrocarvone (with low incidence of unsought side reactions) by using whole-cells of Cryptococcusgastricus. A factorial design (2e5) including five independent variables was performed: X1 = incubationtime; X2 = pH; X3 = amount of whole-cells; X4 = concentration of (4S)-(+)-carvone; X5 = concentration ofcofactor-recycling system. The utilization of glucose and glycerol as cofactor-recycling systems waschecked. On the basis of the results of factorial design, three independent variables (X1, X3 and X4) outof five were further selected for performing a central composite design (CCD).First and second order polynomial equations obtained by CCD were used to select the optimal values ofindependent variables in order to maximize the bioreduction yield of (4S)-(+)-carvone and, at the sametime, to minimize the occurrence of side reactions (i.e. further reduction of dihydrocarvone todihydrocarveol).

2012 - The resveratrol-analogue 4,4’-dihydroxy-trans-stilbene suppresses transformation in normal mouse fibroblast and inhibits proliferation and invasion of human breast cancer cells [Articolo su rivista]
C., Maccario; M., Savio; D., Ferraro; L., Bianchi; R., Pizzala; L., Pretali; Forti, Luca; L. A., Stivala
abstract

4,4′-dihydroxy-trans-stilbene (DHS) is a synthetic analog of resveratrol, a phytoalexin known for its biological activities. We previously demonstrated that DHS exerts an antiproliferative effect on normal human fibroblasts that is higher than that of the natural parent molecule. No evidence regarding its role in human cancer cell lines has been found thus far. In this study, we investigated the effects of DHS both on chemical-induced transformation of BALB/c 3T3 mouse fibroblasts and on the proliferation and invasion of human breast cancer MCF-7 cells. The results showed that DHS markedly suppresses the two-stage (3-methylcholanthrene plus 12-O-tetradecanoylphorbol-13-acetate) cell transformation. Compared with resveratrol, DHS inhibited both anchorage-dependent and -independent MCF-7 growth more efficiently. In addition, a reduction in the number of cells in S-phase, characterized by a concomitant increase in the levels of p21 and p53 proteins, together with a strong inhibition of pRb protein phosphorylation, was observed in DHS-treated cells. Furthermore, DHS effected a strong reduction in matrix metalloproteinase-2 and -9 activities, concomitantly with a marked inhibition of cell adhesion to the extracellular matrix components as well as inhibition of cell migration and invasion. Importantly, modulation of the adhesion molecule E-cadherin was also found in DHS-treated cells. Taken together, these results demonstrate that the two 4,4′-hydroxyl groups on the stilbenic backbone make DHS a more active molecule than resveratrol in inhibiting neoplastic transformation, cancer cell proliferation and invasion. In conclusion, this study suggests that DHS could be a promising anticancer agent.

2011 - A defective ABC transporter of the MRP family, responsible for the bean lpa1 mutation, affects the regulation of the phytic acid pathway, reduces seed myo-inositol and alters ABA sensitivity [Articolo su rivista]
D., Panzeri; E., Cassani; E., Doria; G., Tagliabue; Forti, Luca; B., Campion; R., Bollini; C. A., Brearley; R., Pilu; E., Nielsen; F., Sparvoli
abstract

• We previously identified the lpa1 (low phytic acid) 280-10 line that carries amutation conferring a 90% reduction in phytic acid (InsP6) content. In contrast toother lpa mutants, lpa1(280-10) does not display negative pleiotropic effects. Inthe present paper, we have identified the mutated gene and analysed its impact onthe phytic acid pathway.• Here, we mapped the lpa1(280-10) mutation by bulk analysis on a segregatingF2 population, an then, by comparison with the soybean genome, we identifiedand sequenced a candidate gene. The InsP6 pathway was analysed by geneexpression and quantification of metabolites.• The mutated Pvmrp1(280-10) cosegregates with the lpa1(280-10) mutation,and the expression level of several genes of the InsP6 pathway are reduced in thelpa1(280-10) mutant as well as the inositol and raffinosaccharide content.PvMrp2, a very similar paralogue of PvMrp1 was also mapped and sequenced.• The lpa1 mutation in beans is likely the result of a defective Mrp1 gene (orthologousto the lpa genes AtMRP5 and ZmMRP4), while its Mrp2 paralog is not ableto complement the mutant phenotype in the seed. This mutation appears todown-regulate the InsP6 pathway at the transcriptional level, as well as alteringinositol-related metabolism and affecting ABA sensitivity.

2011 - Bioreduction of α,β-unsatured ketones and aldehydes by non-conventional yeast (NCY) whole-cells [Articolo su rivista]
M., Goretti; C., Ponzoni; E., Caselli; E., Marchigiani; Cramarossa, Maria Rita; B., Turchetti; Forti, Luca; P., Buzzini
abstract

The bioreduction of alfa,beta-unsaturated ketones (ketoisophorone, 2-methyl- and 3-methyl-cyclopentenone)and aldehydes [(S)-(-)-perillaldehyde and alfa-methyl-cinnamaldehyde] by 23 ‘‘non-conventional’’ yeasts (NCYs) belonging to 21 species of the genera Candida, Cryptococcus, Debaryomyces, Hanseniaspora, Kazachstania, Kluyveromyces, Lindnera, Nakaseomyces, Vanderwaltozyma, and Wickerhamomyces was reported. The results highlight the potential of NCYs as whole-cell biocatalysts for selective biotransformation of electron-poor alkenes. A few NCYs exhibited extremely high (>90%) or even total ketoisophorone and 2-methyl-cyclopentenone bioconversion yields via asymmetric reduction of the conjugated C=C bond catalyzed by enoate reductases. Catalytic efficiency declined after switching from ketones to aldehydes. High chemoselectivity due to low competing carbonyl reductases was also sometimes observed.

2011 - Structure-Activity Relationship of Resveratrol and Its Analogue, 4,4 '-Dihydroxy-Trans-Stilbene, Toward the Endothelin Axis in Human Endothelial Cells [Articolo su rivista]
T., Coppa; M. C., Lazze; O., Cazzalini; P., Perucca; R., Pizzala; L., Bianchi; L. A., Stivala; Forti, Luca; C., Maccario; V., Vannini; M., Savio
abstract

Resveratrol inhibits endothelin-1, a vascular tension regulator. We synthesized the resveratrol analogue 4,4'-dihydroxy-trans-stilbene with 2 hydroxyl groups in the 4 and 4' position to obtain a molecule more active than resveratrol (3,4',5-trihydroxy-trans-stilbene). The results demonstrate that 4,4'-dihydroxy-trans-stilbene led to a significant decrease in total endothelin-1 secretion and in endothelin-1 messenger RNA (mRNA) levels in human endothelial cells. In addition, resveratrol and its analogue decreased endothelin-converting enzyme-1 mRNA levels and further reduced the activity of the enzyme. 4,4'-dihydroxy-trans-stilbene was more active than resveratrol because the new molecule exerted greater activity at the level of endothelin synthesis and conversion, even at a lower concentration. Although 4,4'-dihydroxy-trans-stilbene and resveratrol inhibited formation of reactive oxygen species and lipid peroxidation, the treatment of cells with different oxidant agents did not modify the endothelin-1 release. This finding suggests that the inhibition of endothelin-1 secretion is independent of the antioxidant properties of the 2 compounds. On the basis of these results, the resveratrol analogue 4,4'-dihydroxy-trans-stilbene could be a promising chemopreventive agent against cardiovascular diseases.

2010 - Rapid method for screening enoate reductase activity in yeasts [Articolo su rivista]
Raimondi, Stefano; Roncaglia, Lucia; Amaretti, Alberto; Leonardi, Alan; P., Buzzini; Forti, Luca; Rossi, Maddalena
abstract

Old Yellow Enzymes (OYEs, EC 1.6.99.1) are flavin-dependent oxidoreductases that catalyze the asymmetric reduction of electron-poor alkenes (enoate reductase activity). Since OYEs are involved in detoxification of acrolein, a high-throughput method for selecting yeasts expressing high enoate reductase activity, based on their acrolein resistance, was developed. The screening method was based on the measurement of growth in acrolein-supplemented medium, in 96-well microtiter plate cultures. A quantitative descriptor (Acrolein Resistance Factor = ARF) was firstly designed for quantifying the influence of both acrolein concentration and time of exposure on yeast growth. Besides, the efficiency of bioconversion of ketoisophorone (KIP) was exploited to measure OYE activity. In order to validate the method, ARF was correlated with the bioconversion of KIP on thirty yeast strains, belonging to 7 genera. With only a few exceptions, all strains exhibiting the highest ARF also displayed the maximum OYE activity. The presence of OYE genes in the strains showing OYE activity was confirmed by PCR amplification. Based on the results herein reported, this method should be profitably used as a fast screening procedure aimed at selecting outstanding strains for whole-cell bioconversions and could open many possibilities for the isolation and the biocatalytic exploitation of new OYEs from yeasts.

2009 - Biotransformation of electron-poor alkenes by yeasts: asymmetric reduction of (4S)-(+)-carvone by yeast enoate reductases [Articolo su rivista]
M., Goretti; Ponzoni, Chiara; Caselli, Elisa; E., Marchigiani; Cramarossa, Maria Rita; B., Turchetti; P., Buzzini; Forti, Luca
abstract

Within the framework of a large-scale screening carried out on 146 yeasts of environmental origin, 16 strains (11% of the total) exhibited the ability to biotransform (4S)-(+)-carvone. Such positive yeasts, belonging to 14 species of 6 genera (Candida, Cryptococcus, Hanseniaspora, Kluyveromyces, Pichia andSaccharomyces), were thus used under different physiological state (growing, resting and lyophilised cells). Yields (expressed as% of biotransformation) varied from 0.14 to 30.04%, in dependence of both the strain and the physiological state of the cells. Products obtained from reduction of (4S)-(+)-carvone were1S,4S- and 1R,4S-dihydrocarvone, (1S,2S,4S)-, (1S,2R,4S)- and (1R,2S,4S)-dihydrocarveol. Only traces of (1R,2R,4S)-dihydrocarveol were observed in a few strains. As far as the stereoselectivity of the biocatalysis, with the sole exception of a few strains, the use of yeasts determined the prevalent accumulation of 1S,4S-isomers [(1S,4S)-dihydrocarvone + (1S,2S,4S)-dihydrocarveol + (1S,2R,4S)-dihydrocarveol].The addition of glucose (acting as auxiliary substrate for cofactor-recycling system) to lyophilised yeast cells determined a considerable increase of biocatalytic activity: in particular, two strains showed a surprising increase of the% of biotransformation of (4S)-(+)-carvone (to values >98%).

2009 - Psychrophilics yeasts: new biocatalytic agents for enoate reductase activities [Abstract in Rivista]
E., Marchegiani; M., Goretti; Caselli, Elisa; E., Branda; Cramarossa, Maria Rita; Ponzoni, Chiara; B., Turchetti; Forti, Luca; P., Buzzini
abstract

Owing to widespread diffusion of areas characterized by permanent low temperatures (e.g. deep-sea waters, mountains and polar regions), psychrophilic microorganisms have the largest terrestrial distribution. Adaptation of microbial life at low temperatures requires some changes, mainly at the level of cell membrane components (including transport systems), proteins and nucleic acids.Psychrophilic organisms produce enzymes functioning in coldenvironment and simply denaturing at high temperature. Suchcold-active enzymes have a more flexible structure to undergo the conformational changes necessary for catalysis with a lower energy demand. Owing to their high catalytic activity at low temperatures, low thermostability and unusual specificities, cold-active enzymes offer novel opportunities for biotechnological exploitation.The enormous potential of biocatalysts for the production of enantiopure products of industrial interest remains to beexploited, especially in the area of enantioselective alkene reductions, due to their marked regio- and stereo-selectivity. One of the emerging reductive chemistry is the asymmetric reductionof alpha,beta-unsaturated carbonyl compounds through microbialwhole-cells rich in enoate reductase (ER), a flavin-dependentenzyme belonging to the ‘old yellow enzyme’ (OYE) family.Even though massive screenings for yeasts as reducing catalystshave been carried out so far, there is still a need for new biocatalysts able to performing highly stereoselective reactions. Consequently, the potential of yeast biocatalysis appears to be still far from being fully explored. In this study we report the results of a screening on facultative psychrophilic yeasts (belonging to the species Cryptococcus gilvescens and Cryptococcus gastricus) for new cold-adapted enoate reductases using (4S)-(+)-carvone as a model substrate. The preliminary results obtained using growing cells demonstrate that the ERs activities are apparently expressed to a greater extent inpsychrophilic yeasts with respect to the mesophilic ones. In particular, C. gastricus exhibited the better biocatalytic performances.The results obtained by using other catalytic forms (resting orlyophilized cells) will be presented.

2009 - Structure–activity relationships of resveratrol and derivatives in breast cancer cells [Articolo su rivista]
R., Lappano; C., Rosano; A., Madeo; L., Albanito; P., Plastina; B., Gabriele; Forti, Luca; L. A., Stivala; D., Iacopetta; V., Dolce; S., Andò; V., Pezzi; M., Maggiolini
abstract

Resveratrol (RSV) is classified as a phytoestrogen due to its ability to interact with estrogen receptors (ERs). We assessed structure–activity relationships of RSV and the analogs 4,4'-dihydroxystilbene (4,4'-DHS), 3,5-dihydroxystilbene (3,5-DHS), 3,4'-dihydroxystilbene (3,4'-DHS), 4-hydroxystilbene (4-HS) using as model systems the ERa-positive and negative MCF7 and SkBr3 breast cancer cells, respectively. In binding assays and transfection experiments RSV and the analogs showed the following order of agonism for ERa: 3,4'-DHS > 4,4'-DHS > 4-HS > RSV, while 3,5-DHS did not elicit any ligand properties. Computational docking analysis and real-time PCR revealed for each analog a distinct ERa binding orientation and estrogen target gene expression profile. Interestingly, the aforementionedorder of ligand activity was confirmed in proliferation assays which also showed the lack of growth stimulation by 3,5-DHS. Our data suggest that subtle changes in the structure of the RSVderivatives examined may be responsible for the different ERa-mediated biological responses observed in estrogen-sensitive cancer cells.

2009 - The resveratrol analogue 4,4-dihydroxy-trans-stilbene inhibits cell proliferation with higher efficiency but different mechanism from resveratrol [Articolo su rivista]
M., Savio; T., Coppa; L., Bianchi; V., Vannini; G., Maga; Forti, Luca; O., Cazzalini; M. C., Lazzè; P., Perucca; E., Prosperi; L. A., Stivala
abstract

Resveratrol (3,4',5-trihydroxy-trans-stilbene) is a natural phytoalexin found in grapes and wine, which shows antiproliferative activity. We previously found that 4-hydroxy group in the trans conformation was absolutely required for the inhibition of cell proliferation. In the present work we have synthesized the resveratrol analogue 4,4'-dihydroxy-trans-stilbene, which contains two OH in 4' and 4 positions,with the aim of developing a compound with an antiproliferative potential higher than that of resveratrol, on the basis of the correlation between structure and activity previously observed. In comparison with resveratrol, 4,4'-dihydroxy-trans-stilbene inhibited cell clonogenic efficiency of fibroblasts ninetimes more although with a different mechanism. First, 4,4'-dihydroxy-trans-stilbene induced predominantly an accumulation of cells in G1 phase, whereas resveratrol perturbed the G1/S phase transition. Second, although both compounds were able to inhibit DNA polymerase (pol) delta in an in vitro assay, 4,4'-dihydroxy-trans-stilbene did not affect pol alfa activity. Finally, 4,4'-dihydroxy-trans-stilbene increased p21CDKN1A and p53 protein levels, whereas resveratrol led to phosphorylation of the S-phase checkpoint protein Chk1. Taken together, our results demonstrated for the first time that the two hydroxyl groups on4- and 4'- positions of the stilbenic backbone enhance the antiproliferative effect and introduce additional targets in the mechanism of action of resveratrol. In conclusion, 4,4'-dihydroxy-trans-stilbene has potent antiproliferative activities that differ from the effect of resveratrol shown in this system, suggesting that it warrants further development as a potential chemopreventive or therapeutic agent.

2008 - Biotransformation of Acyclic Monoterpenoids by Debaryomyces sp., Kluyveromyces sp. and Pichia sp. Strains of Environmental Origin [Articolo su rivista]
Ponzoni, Chiara; C., Gasparetti; M., Goretti; B., Turchetti; Pagnoni, Ugo Maria; Cramarossa, Maria Rita; Forti, Luca; P., Buzzini
abstract

Sixty yeast strains, which belong to 32 species of the genera Debaryomyces, Kluyveromyces and Pichia, and which were isolated from plant-, soil- or insect-associated habitats, were screened for their ability to biotransform the acyclic monoterpenes geraniol and nerol. The aptitude to convert both compounds (from 2.6 to 30.6 and from 2.7 to 29.1 %/g cell DW, = dry weight), resp.) was apparently a broad distributed character in such yeasts. Depending upon the substrate used, the production of linalool, alfa-terpineol, β-myrcene, D-limonene, (E)-β-ocimene, (Z)-β-ocimene or carene was observed. Linalool was the main product obtained from geraniol, whereas linalool and alfa-terpineol were the main products obtained through the conversion of nerol. Yet, differently from nerol, the aptitude to exhibit high bioconversion yields of geraniol to linalool was an apparently genus-related character, whereas the ability to produce other monoterpenes was a both genus- and habitat-related character. The possible pathways of bioconversion of geraniol or nerol to their derivative were proposed/discussed.

2008 - Resveratrol, a natural antioxidant phytoalexin, and 4 synthetic derivatives can promote expression of gamma-globin gene in vitro [Abstract in Rivista]
P., Spyrou; Forti, Luca; L., Kithreotis; A., Kirri; S., Christou; L. A., Stivala; M., Kleanthous
abstract

Resveratrol (3,4′5-trihydroxystilbene) is a naturally occurring phenolic compound. It is found in the skin of red grapes and in a variety of medicinal plants. Resveratrol presents many biological activities that have been applied against cardiovascular disease and cancer. One of Resveratrol properties is inhibition of the enzyme Ribonucleotide reductase. It has been found that some other Ribonucleotide reductase inhibitors like Hydroxyurea, Didox and Trimidox are also inducers of gammaglobin gene expression. In this study we investigated whether Resveratrol and 10 newly synthesized derivatives can act as inducers of gamma-globin gene and increase HbF levels. These derivatives were synthesized by the group of Dr. L.A Stivala (Department of Chemistry, University of Modena) by modifications of the polar groups of Resveratrol structure in an effort to increase its antioxidant activity. We examined the effect of Resveratrol and 10 of its derivatives on gamma- and beta-globin gene promoter activity in a dual luciferase assay in GM979 cells. Hydroxyurea, a known Ribonucleotide reductase inhibitor and HbF inducer and propionic acid, an HbF inducer, were used as positive control substances. The results showed that Resveratrol and 4 of its derivatives increased the gamma-globin gene promoter activity more than 3 fold. The compounds which exhibited an effect were subsequently tested in human erythroid liquid cultures derived from normal individuals and thalassemic patients. HbF content was measured by HPLC. Despite their promising effects on gamma-globin gene promoter activity in MEL cells, Resveratrol and its 4 active derivative compounds did not cause significant increase in HbF production in erythroid progenitor cells (BFUe).

2008 - αvβ3 Integrin-dependent antiangiogenic activity of resveratrol stereoisomers [Articolo su rivista]
M., Belleri; D., Ribatti; M., Savio; L. A., Stivala; Forti, Luca; E., Tanghetti; P., Alessi; D., Coltrini; A., Bugatti; S., Mitola; S., Nicoli; V., Vannini; M., Presta
abstract

Angiogenesis is target for antineoplastic and chemopreventivetherapies. The natural phytoalexin resveratrolis found in grapes and red wine as cis and trans stereoisomers.trans-Resveratrol shows antiangiogenic activity,but its mechanism of action is not fully elucidated.Recently, trans-resveratrol has been shown to interactwith the B3 integrin subunit, raising the possibility thatinhibition of endothelial AvB3 integrin function may concurto its angiosuppressive activity. To get novel insightsabout the antiangiogenic activity of resveratrol, we comparedcis- and trans-resveratrol stereoisomers for theireffect on the angiogenesis process and endothelial AvB3integrin function. trans-Resveratrol inhibits endothelialcell proliferation and the repair of mechanically woundedendothelial cell monolayers. Also, it prevents endothelialcell sprouting in fibrin gel, collagen gel invasion, andmorphogenesis on Matrigel. In vivo, trans-resveratrolinhibits vascularization of the chick embryo area vasculosaand murine melanoma B16 tumor growth and neovascularization.In all the assays, cis-resveratrol exerts a limited, if any, effect. In keeping with these observations, transresveratrol,but not cis-resveratrol, inhibits AvB3 integrindependentendothelial cell adhesion and the recruitmentof enhanced green fluorescent protein-tagged B3 integrinin focal adhesion contacts. In conclusion, stereoisomeryaffects the antiangiogenic activity of resveratrol, the transisomer being significantly more potent than the cis isoform.The different antiangiogenic potential of resveratrolstereoisomers is related, at least in part, to theirdifferent capacity to affect AvB3 integrin function. Thismay have profound implications for the design of syntheticantiangiogenic/angiopreventive phytoalexin derivatives.

2007 - Laccase catalyzed dimerization of hydroxystilbenes [Articolo su rivista]
Ponzoni, Chiara; E., Beneventi; Cramarossa, Maria Rita; Raimondi, Stefano; G., Trevisi; Pagnoni, Ugo Maria; S., Riva; Forti, Luca
abstract

A series of hydroxystilbenes, analogues of the bioactive phytoalexin resveratrol, were synthesized and submitted to the catalytic action of a laccase from Trametes pubescens in a biphasic system made of ethyl acetate and acetate buffer. Oxidation took place at the 4’-hydroxy (4-hydroxy) position ofthe hydroxystilbenicm oieties, followed by radicalradicalcoupling dimerization reactions. Most of the products were isolated in good yields and fully characterized.Depending on the substrates, three different dimeric products could be identified, the main products usually being 4-O-a-ß-5 (dihydrofuran-like) dimers.

2006 - Anthocyanidins decrease endothelin-1 production and increase endothelial nitric oxide synthase in human endothelial cells [Articolo su rivista]
M. C., Lazzè; R., Pizzala; P., Perucca; O., Cazzalini; M., Savio; Forti, Luca; V., Vannini; L., Bianchi
abstract

Epidemiological and intervention studies correlate anthocyanin-rich beverages and a low incidence of coronary heart diseases, Since endothelin-1 (ET-1) and nitric oxide (NO) produced by endothelial NO synthase (eNOS) are vascular tension regulators secreted by endothelial cells, we studied the influence of two anthocyanidins, namely cyanidin (CY) and delphinidin (DP), on the regulation of ET-I and eNOS in cultured human umbilical vein endothelial cells (HUVECs). Aglycon anthocyanidin forms, such as CY and DP, may be present in vivo after the first deglycosylation step occurring in the jejunum and in the liver. DP showed a major action compared to CY inducing a significant dose-dependent inhibitory effect on both protein and mRNA levels of ET-1. CY and DP both increased the protein level of eNOS, but DP showed the major effect raising eNOS protein in a dose-dependent manner. To correlate the vasoprotective effect of CY and DP with their antioxidant activity, we analysed also the antioxidant effect of anthocyanidins both in vitro and in HUVECs. In particular, we examined the effect of anthocyanidins on endothelial heme oxygenase-1 (HO-1), an inducible stress protein. In all tests, DP showed a higher antioxidant activity than CY. Finally, the antiproliferative effect induced by DP was detected in HUVECs. DP and CY differ in the number and position of hydroxyl groups in their structure;, therefore, the greater biological activity by DP, compared with CY, seems to be due to the presence of the three hydroxyl groups on the B ring in the molecular structure of DP.

2006 - Application of the response surface methodology (RSM) for optimizing the production of volatile organic compounds (VOCs) by Trichosporon moniliiforme [Articolo su rivista]
C., Gasparetti; P., Buzzini; Cramarossa, Maria Rita; B., Turchetti; Pagnoni, Ugo Maria; Forti, Luca
abstract

Response surface methodology (RSM) was applied in optimizing the production of volatile organic compounds (VOCs) by shake-flask cultures of a selected Trichosporon moniliiforme strain (DBVPG 4665). A factorial experiment (2(3)) was designed by setting three parameters at two levels: X-1 = incubation time (IT = 72-168 h); X-2 = pH (4-7); X-3 = L-methionine concentration (L-MET = 5-20 g l(-1)). On the basis of the results of factorial design, X-2 (pH) and X-3 (L-MET) were optimized by performing a central composite design (CCD). Second-order polynomial models and response surfaces were obtained for total VOC concentration (VOC1) and for the percentage of dimethyl disulphide (DMDS)/total VOCs (%DMDS). A range of variation of X-2 and X-3 able to maximize simultaneously both responses has been found. Average final values of responses (VOC1 = 4.9 +/- 1.3 mg l(-1); %DMDS = 97.6 +/- 1.1%) were about 532 and 133% of data of central point and were in good agreement with calculated values, thus confirming the reliability of our empirical model in optimizing VOCs production by T. moniliiforme (c) 2006 Elsevier Inc. All rights reserved.

2006 - Industrial applications of culture collections: yeasts of environmental origin as cellar biocatalysts for the biotransformation of monoterpenoids of agroalimentary interest [Relazione in Atti di Convegno]
Buzzini, ; C., Gasparetti; Cramarossa, Maria Rita; B., Turchetti; Forti, Luca; A., Vaughan Martini
abstract

2005 - Antiangiogenic and vascular-targeting activity of the microtubule-destabilizing trans-resveratrol derivative 3,5,4 '-trimethoxystilbene [Articolo su rivista]
M., Belleri; D., Ribatti; S., Nicoli; F., Cotelli; Forti, Luca; V., Vannini; L. A., Stivala; M., Presta
abstract

Neovascularization plays an important role in neoplasia and angioproliferative diseases. Two major modalities have been developed so far to affect neovascularization: its prevention by antiangiogenic compounds, and immature vessel disruption by vascular-targeting agents. trans-Resveratrol, found in grapes and wine, exerts antioxidant, antineoplastic, and antiangiogenic activities. Here, among various synthetic trans-resveratrol derivatives tested, 3,5,4'-trimethoxystilbene was an antiangiogenic agent 30 to 100 times more potent than parent compound in inhibiting endothelial cell proliferation, sprouting, collagen gel invasion, and morphogenesis (ID50 = 0.3-3.0 mu M). In addition, 3,5,4'-trimethoxystilbene acts as a vascular-targeting agent by causing microtubule disassembling and tubulin depolymerization and by impairing the repositioning of the microtubule organization center and the formation of membrane ruffles in migrating endothelial cells. In keeping with a vascular-targeting ability, 3,5,4'-trimethoxystilbene induced apoptosis only in subconfluent endothelial cells and apoptotic regression of immature vessels in the ex vivo rat aorta ring assay. In vivo, 3,5,4'-trimethoxystilbene caused the rapid stasis of blood flow and regression of intersegmental vessels in the trunk of zebrafish embryos. In addition, it inhibited blood vessel growth and caused the disappearance of pre-existing blood vessels in the area vasculosa of the chick embryo. In conclusion, 3,5,4'-trimethoxystilbene associates an antiangiogenic profile to a significant vascular-targeting activity.

2005 - Biocatalytic reduction of (+)- and (–)-carvone by bacteria [Articolo su rivista]
Cramarossa, Maria Rita; A., Nadini; Bondi, Moreno; Messi, Patrizia; Pagnoni, Ugo Maria; Forti, Luca
abstract

The biotransformation of the two enantiomers of carvone, (–)-(1a) and (+)-(1b), by two bacteria (Pseudomonas putida and Acinetobacter lwoffi) isolated from different environments is reported. The metabolites from the biotransformations carried out in liquid cultures using the sterile TS-2 mineral medium were screened by headspace solid phase microextraction/GC–MS. The results show that these bacteria are efficient tools in the diastereoselective bioreduction of (+)- and (–)-carvone.

2005 - Enantioselective esterase activity of an industrial glutaryl acylase [Articolo su rivista]
S., Adani; Raimondi, Stefano; Forti, Luca; D., Monti; S., Riva
abstract

The unexpected esterase activity of an industrial glutaryl acylase was investigated. Glutaryl esters of a series of primary and secondary alcohols as well as of phenols were all efficiently hydrolyzed, the only exception being the sterically hindered glutarate of thymol. The enantioselectivities of the acylase, which were evaluated with three of these substrates, were quite low (E values ranging between 1.9 and 7.2), but were significantly improved by substrate and/or solvent engineering. Enantiomerically enriched hydrolyzed alcohols and unreacted glutarates can be easily separated by selective extraction, thus avoiding chromatographic steps.

2005 - Inhibition of mammalian DNA polymerases by resveratrol: mechanism and structural determinants [Articolo su rivista]
G. A., Locatelli; M., Savio; Forti, Luca; I., Shevelev; K., Ramadan; L. A., Stivala; V., Vannini; U., Hubscher; S., Spadari; G., Maga
abstract

Resveratrol, a natural compound found in many dietary plants and in red wine, plays an important role in the prevention of many human pathological processes, including inflammation, atherosclerosis and carcinogenesis. We have shown that the antiproliferative activity of resveratrol correlated with its ability to inhibit the replicative pols (DNA polymerases) alpha and delta in vitro [Stivala, Savio, Carafoli, Perucca, Bianchi, Maga, Forti, Pagnoni, Albini, Prosperi and Vannini (2001) J. Biol. Chem. 276,22586-22594]. In this paper, we present the first detailed biochemical investigation on the mechanism of action of resveratrol towards mammalian pols. Our results suggest that specific structural determinants of the resveratrol molecule are responsible for selective inhibition of different mammalian pols, such as the family B pot a and the family X pot X. Moreover, the resveratrol derivative trans-3,5-dimethoxy-4-hydroxystilbene, which is endowed with a strong antiproliferative activity (Stivala et al., 2001), can inhibit pols a and; and also suppress the in vitro SV40 DNA replication. The potency of inhibition is similar to that of aphidicolin, an inhibitor of the three replicative pols alpha, delta and epsilon. Our findings establish the necessary background for the synthesis of resveratrol derivatives having more selective and potent antiproliferative activity.

2005 - Production of volatile organic compounds (VOCs) by yeasts isolated from the ascocarps of black (Tuber melanosporum Vitt.) and white (Tuber magnatum Pico) truffles [Articolo su rivista]
P., Buzzini; C., Gasparetti; B., Turchetti; Cramarossa, Maria Rita; A., Vaughan Martini; A., Martini; Pagnoni, Ugo Maria; Forti, Luca
abstract

Twenty-nine yeast strains were isolated from the ascocarps of black and white truffles (Tuber melanosporum Vitt. and Tuber magnatum Pico, respectively), and identified using a polyphasic approach. According to the conventional taxonomic methods, MSP-PCR fingerprinting and sequencing of the D1/D2 domain of 26S rDNA, the strains were identified as Candida saitoana, Debaryomyces hansenii, Cryptococcus sp., Rhodotorula mucilaginosa, and Trichosporon moniliiforme. All isolates assimilated L-methionine as a sole nitrogen source and produced the volatile organic compounds (VOCs), 2-methyl butanol, 3-methyl butanol, methanethiol, S-methyl thioacetate, dimethyl sulfide, dimethyl disulfide, dimethyl trisulfide, dihydro-2-methyl-3(2H)-thiophenone and 3-(methylthio)-1-propanol (MTP). ANOVA analysis of data showed significant (P < 0.01) differences in VOCs produced by different yeasts, with MTP as the major component (produced at concentrations ranging from 19.8 to 225.6 mg/l). In addition, since some molecules produced by the isolates of this study are also characteristic of truffle complex aroma, it is possible to hypothesize a complementary role of yeasts associated with this ecosystem in contributing to final Tuber spp. aroma through the independent synthesis of yeast-specific volatile constituents.

2004 - Biotransformation of resveratrol: synthesis of trans-dehydrodimers catalyzed by laccases from Myceliophtora thermophyla and from Trametes pubescens [Articolo su rivista]
S., Nicotra; Cramarossa, Maria Rita; Mucci, Adele; Pagnoni, Ugo Maria; S., Riva; Forti, Luca
abstract

Laccases from different sources have been used, for the first time, for the selective oxidation of the stilbenic phytoalexin transresveratrol (3,5,4´-trihydroxystilbene, 1a) on a preparative scale. Specifically, the enzymes from Myceliophtora thermophyla and from Trametes pubescens gave the dehydrodimer 2a in 31 and 18% isolated yields, respectively. These results compare favorably with the reported data for the chemically catalyzed dimerization of la (18% yields). The antioxidant properties of 2a have also been investigated.

2004 - Produzione di aromi e fragranze da impiegarsi in campo alimentare mediante biodegradazioni microbiche di terpeni [Relazione in Atti di Convegno]
Bondi, Moreno; Cramarossa, Maria Rita; Forti, Luca; A., Nadini; Pagnoni, Ugo Maria; Messi, Patrizia
abstract

2004 - Resveratrol is a peroxidase-mediated inactivator of COX-1 but not COX-2 - A mechanistic approach to the design of COX-1 selective agents [Articolo su rivista]
L. M., Szewczuk; Forti, Luca; L. A., Stivala; T. M., Penning
abstract

Resveratrol (3,4', 5- trihydroxy- trans- stilbene) is a phytoalexin found in grapes that has anti- inflammatory, cardiovascular protective, and cancer chemopreventive properties. It has been shown to target prostaglandin H-2 synthase (COX)-1 and COX-2, which catalyze the first committed step in the synthesis of prostaglandins via sequential cyclooxygenase and peroxidase reactions. Resveratrol discriminates between both COX isoforms. It is a potent inhibitor of both catalytic activities of COX-1, the desired drug target for the prevention of cardiovascular disease, but only a weak inhibitor of the peroxidase activity of COX-2, the isoform target for nonsteroidal anti- inflammatory drugs. We have investigated the unique inhibitory properties of resveratrol. We find that it is a potent peroxidase- mediated mechanism-based inactivator of COX-1 only (k(inact) = 0.069 +/- 0.004 s(-1), K-i( inact) = 1.52 +/- 0.15 muM), with a calculated partition ratio of 22. Inactivation of COX-1 was time- and concentration- dependent, it had an absolute requirement for a peroxide substrate, and it was accompanied by a concomitant oxidation of resveratrol. Resveratrol-inactivated COX-1 was devoid of both the cyclooxygenase and peroxidase activities, neither of which could be restored upon gel- filtration chromatography. Inactivation of COX-1 by [H-3] resveratrol was not accompanied by stable covalent modification as evident by both SDS-PAGE and reverse phase- high performance liquid chromatography analysis. Structure activity relationships on methoxy- resveratrol analogs showed that the m-hydroquinone moiety was essential for irreversible inactivation of COX-1. We propose that resveratrol inactivates COX-1 by a hit- and- run mechanism, and offers a basis for the design of selective COX-1 inactivators that work through a mechanism- based event at the peroxidase active site.

2003 - Biodegradazioni microbiche di terpeni per la produzione di aromi e fragranze [Relazione in Atti di Convegno]
Bondi, Moreno; Cramarossa, Maria Rita; Forti, Luca; Nadini, A; Pagnoni, Ugo Maria; Messi, Patrizia
abstract

2003 - Biotrasformazioni di terpeni mediate da batteri ambientali [Abstract in Atti di Convegno]
Bondi, Moreno; Cramarossa, Maria Rita; Forti, Luca; Messi, Patrizia; A., Nadini; Pagnoni, Ugo Maria
abstract

Negli ultimi anni si è avuto un incremento della produzione biotecnologica di aromi chimici naturali (NACs) legato alla richiesta, da parte dei consumatori, di prodotti biologici. L’interesse per gli aromi naturali rispetto a quelli ottenuti per sintesi chimica ha comportato un aumento della produzione “microbica” di bioaromi. Un aroma per essere definito “naturale” deve essere ottenuto da sostanze naturali attraverso trasformazioni biocatalitiche che utilizzino enzimi o microrganismi ed il prodotto finale deve essere identico ad una sostanza nota già presente in natura. Se lo stesso prodotto è ottenuto per sintesi chimica si può classificare come “natural identic”. I terpenoidi sono precursori naturali poco costosi, facilmente reperibili e rinnovabili e costituiscono il maggior gruppo di prodotti naturali con piu’ di 22.000 strutture conosciute. I monoterpeni, che sono ampiamente distribuiti in natura e sono fra i componenti principali degli olii essenziali di erbe, spezie e conifere, costituiscono una classe di substrati precursori molto utili e, dato il loro utilizzo nell’industria profumiera ed alimentare, sono prodotti di partenza ideali per la produzione biotecnologica di NACs.Sono note in letteratura diverse bioconversioni di monoterpeni per la produzione di aromi naturali con vari microorganismi, in particolare funghi e lieviti, mentre sono state meno sudiate le bioconversioni con batteri. In questo lavoro sono riportate le biotrasformazioni di citrale e carvone biocatalizzate da alcuni batteri ambientali: Pseudomonas putida, Pseudomonas aeruginosa, Pseudomonas cepacia, Pseudomonas fluorescens, Acinetobacter lwoffi e Flavobacterium spp. In tutte le bioconversioni le uniche fonti di carbonio e di energia sono rappresentate dai monoterpeni. Pseudomonas putida riduce selettivamente il carvone a diidrocarvone, mentre Acinetobacter lwoffi, Flavobacterium spp. e Pseudomonas aeruginosa sono in grado di ridurre ulteriormente il prodotto a diidrocarveolo. Non vi è alcuna bioconversione né con la Pseudomonas cepacia nè con la Pseudomonas fluorescens. La citrale (miscela di nerale e geraniale) è ridotta a nerolo e geraniolo da Acinetobacter lwoffi mentre con Pseudomonas putida si ha un’ulteriore riduzione a citronellolo. La bioconversione con Flavobacterium spp è caratterizzata da un cammino riduttivo, che porta dapprima alla formazione di nerolo e geraniolo e successivamente a citronellolo, e da un cammino degradativo, in cui si ottiene il 6-metil-5-epten-2-one (un prodotto lineare di degradazione dei terpeni) nelle prime 24 ore e successivamente CO2 ed H2O. Lo stesso cammino degradativo si ha anche con Pseudomonas cepacia, con Pseudomonas fluorescens e con Pseudomonas aeruginosa.

2003 - Glutaryl-7-ACA acylase: a new tool for the biocatalyzed kinetic resolution of racemic amines and alcohols [Abstract in Rivista]
Raimondi, Stefano; Forti, Luca; D., Monti; S., Riva
abstract

The enzymes D-amino acid oxidase and glutaryl-7-ACAacylase (GA) are currently utilised for the industrial productionof 7-aminocephalosporanic acid (7-ACA, 2), an importantprecursor of semisyntetic cephalosporins. Specifically,GA is devoted to the cleavage of the amide bond betweenglutaric acid and 7-ACA in the intermediate glutaryl-7-ACA(1).The synthetic performances of this enzyme towards 1have been widely investigated, while very little is known onGA substrate specificity.We have found that an industrial GAis very specific for the acyl moiety that has to be released, theglutaryl derivatives being by far the best substrates. On theother hand, this enzyme accepts a wide variety of “leavinggroups”. Not only N-glutarate of b-lactam derivatives, butalso N-glutaryl aminoacids as well as N-glutarylamides (aromaticand aliphatic) could be hydrolysed by GA, which, additionally,showed a significant esterase activity. Morenotably, GA-catalysed hydrolyses were highly enantioselective

2003 - Kinetic resolutions of racemic amines and alcohols catalyzed by an industrial glutaryl-7-aminocephalosporanic acid acylase with unexpected broad substrate specificity [Articolo su rivista]
Raimondi, Stefano; Forti, Luca; D., Monti; S., Riva
abstract

An industrial glutaryl-7-aminocephalosporanic acid acylase (GAR) possesses a significant broad substrate specificity that crosses over the usual cephalosporanic skeleton. Enantioselective amidase and even esterase activities have been observed with all the glutarates of racemic substrates investigated, with a stereopreference for the (S)-enantiomer. The different physical-chemical properties of reagents and products allow their easy separation by solvent extraction, avoiding cumbersome chromatography or distillation processes during reaction work-up. (C) 2003 Elsevier Science Ltd. All rights reserved.

2003 - Oxidative coupling of natural phenol derivatives catalyzed by laccases [Abstract in Rivista]
S., Nicotra; B., Danieli; Forti, Luca; A., Intra; S., Riva
abstract

Laccases (benzenediol:oxygen oxidoreductases; EC 1.10.3.2) are multicopper enzymes that catalyze the one-electron oxidation of a wide variety of substrates, i. e. phenols and aromatic amines, with the concomitant reduction of molecular oxygen to water. Recent reports describe the use of laccases for the oxidation of different classes of organic compounds using suitable “mediators”. Conversely, literature data on the laccase-mediated oxidation of complex natural compounds are quite scant. For instance, 30 years ago a laccase from Polyporus versicolor was used for the oxidation of steroid hormones (i. e. beta-estradiol) in emulsion of water and organic solvents to give a mixture of isomeric dimers.In the frame of our general interest in the biocatalyzed formation of carbon-carbon bonds, we are studying the performances of laccases in the oxidative coupling of natural compounds in water solution or in the presence of organic cosolvents. In this report we will present the results obtained using the laccases from Myceliophtora thermophyla and from Trametes versicolor for the oxidation of the phenolic derivatives beta-estradiol, resveratrol and catechin.

2002 - Mechanism of Inactivation of Prostaglandin H2 Synthase 1 (PGH2S-1) by Resveratrol and its Analogs [Abstract in Rivista]
L., Szewczuk; L. A., Stivala; Forti, Luca; T., Penning
abstract

3,4',5-Trihydroxy-trans-stilbene (resveratrol) is a natural product derived from grapes that has anti-inflammatory, anti-coagulation, and cancer chemopreventive properties. It inhibits both the cyclooxygenase (cox) and peroxidase (perox) activities of Prostaglandin H2 Synthase 1 (PGH2S-1). Resveratrol was rapidly oxidized by the perox activity, but unlike other co-reductants inactivated both enzyme activities. The irreversible loss of cox and perox activity was accompanied by the oxidation of resveratrol to yield the inactivator and had an absolute requirement for a peroxide substrate. Resveratrol represents a new class of inhibitors for PGH2S-1 that inactivate both the cox and perox activities of the enzyme, but require catalytic activation to exert their effects. Perox inhibitors are attractive because catalytic intermediates formed in the perox reaction are potent oxidants, which can generate reactive species that can damage biological macromolecules.

2001 - Cyclization of citronellal to menthone and isomenthone catalyzed by Al/Fe-Pillared Clays [Articolo su rivista]
Cramarossa, Maria Rita; Forti, Luca; Pagnoni, Ugo Maria; M., Vidali
abstract

The cyclization of citronellal to a mixture of menthone and isomenthone (2:1) is catalyzed by Al/Fe-Pillared Clay (Al/Fe-PILC) at 80 degreesC in 1,2-dichloroethane in good yield. At room temperature the products are isopulegol and neo-isopulegol, the isomer ratio depending on the reaction conditions.

2001 - Influenza del tipo di confezione sulla conservazione del patrimonio vitaminico e delle proprietà organolettiche del latte [Articolo su rivista]
M., Marchetti; Forti, Luca
abstract

la ricerca è stata condotta utilizzando un lotto di latte pastorizato di alta qualità, confezionato parte in Tetra Brik, parte in Tetra Rex e parte in bottiglie PET. Dai risultati ottenuti risulta evidente che riducendo l'impatto della luce, attraverso l'impiego di un materiale di confezionamento reso opaco alle radiazioni luminose, o di un multistrato a base cartoncino Tetra Pak, è possibile preservare il patrimonio vitaminico in modo pressochè completo e di conseguenza l'intero suo valore nutrizionale.

2001 - Specific structural determinants are responsible for the antioxidant activity and the cell cycle effects of resveratrol [Articolo su rivista]
L. A., Stivala; M., Savio; F., Carafoli; P., Perucca; L., Bianchi; G., Maga; Forti, Luca; Pagnoni, Ugo Maria; A., Albini; E., Prosperi; V., Vannini
abstract

Resveratrol (3,4',5-trihydroxy-trans-stilbene) is a natural phytoalexin found in grapes and wine, which shows antioxidant and antiproliferative activities. In this study we have investigated whether these properties are dependent on similar or different structural determinants of the molecule. To this purpose, resveratrol derivatives, in which all or each single hydroxylic function were selectively substituted with methyl groups, were synthesized. Analogues with the stilbenic double bond reduced or with the stereoisomery modified were also investigated. The antioxidant activity of these compounds was evaluated by measuring the inhibition of citronellal thermo-oxidation, or the reduction of 2,2 diphenyl-1-picrylhydrazyl radical. In addition, the protection against lipid peroxidation was determined in rat liver microsomes, and in human primary cell cultures. The antiproliferative activity was evaluated by a clonogenic assay, and by analysis of cell cycle progression and DNA synthesis. The results showed that the hydroxyl group in 4' position is not the sole determinant for antioxidant activity. In contrast, the presence of 4'-OH together with stereoisomery in the trans conformation (4'-hydroxystyryl moiety) was absolutely required for inhibition of cell proliferation. Enzymatic assays in vitro demonstrated that inhibition of DNA synthesis was induced by a direct interaction of resveratrol with DNA polymerases alpha and delta.

1999 - Conformationally constrained amino acids: A convenient approach to cis-2,3-methano-GABAs [Articolo su rivista]
Forti, Luca; Ghelfi, Franco; S., Levizzani; Pagnoni, Ugo Maria
abstract

3-Aza-2-oxo-bicyclo[3.1.0]hexanes, which are opened to 2,3-methano-gamma-aminobutyric acids, are obtained in very high yields by a two step procedure from N-protected-N-allyl-alpha-Br-amides. (C) 1999 Elsevier Science Ltd. All rights reserved.

1999 - Easy approach to 3-benzylimino-2-pyrrolidinones from 3-chloro-4-chloromethyl-2-pyrrolidinones [Articolo su rivista]
Ghelfi, Franco; G., Ghirardini; Libertini, Emanuela; Forti, Luca; Pagnoni, Ugo Maria
abstract

3-Benzylimino-2-pyrrolidinones can be prepared in good yield by heating 3-chloro-4-chloromethyl-2-pyrrolidinones, benzylamine and NaI in THF at 80 degrees C. An endo-dehydrohalogenation followed by a SN2' substitution on the intermediate allyl chloride, and finally a shift of the exo-double bond to Delta(3) with attendant tautomerization, appears to be the most probable reaction mechanism. (C) 1999 Elsevier Science Ltd. All rights reserved.

1999 - Rearrangement of N-allyl-alpha,alpha-dichloroamides, beta- or gamma-functionalized, to substituted analogues of the gamma-aminobutyric acid (GABA) [Articolo su rivista]
Bellesia, Franco; Forti, Luca; Ghelfi, Franco; G., Ghirardini; Libertini, Emanuela; Pagnoni, Ugo Maria; Pinetti, Adriano; N., Prochilo
abstract

The rearrangement of gamma-chloro, beta-hydroxy or beta-vinyl N-allyl-N-benzyl-alpha,alpha-dichlorocarboxyamides to gamma-aminobutyric acid analogues is efficiently promoted by CuCl\N,N,N',N'-tetramethylethylendiamine. With the beta-vinyl functionalization a tandem radical-radical reaction, yielding 3-aza-2-oxo-bicyclo[3,3,0]octane adducts, is also observed.

1999 - The influence of benzylic protection and allylic substituents on the CuCl-TMEDA catalyzed rearrangement of N-allyl-N-benzyl-2,2-dihaloamides to gamma-lactams. Application to the stereoselective synthesis of pilolactam. [Articolo su rivista]
Ghelfi, Franco; Bellesia, Franco; Forti, Luca; G., Ghirardini; Grandi, Romano; Libertini, Emanuela; M. C., Montemaggi; Pagnoni, Ugo Maria; Pinetti, Adriano; L., De Buyck; A. F., Parson
abstract

A number of N-benzylic protecting groups and allylic substituents have been investigated for the rearrangement, promoted by CuCl-TMEDA, of N-allyl-2,2-dihaloamides to 3,4-disubstituted gamma-lactams. An appreciable chiral induction was observed at the C-4 site when alpha-phenylethylamine was used as a chiral protecting group, while an unexpected Diels-Alder reaction occurred when using a 2-furyl-methyl protection. This rearrangement has been applied to the synthesis of pilolactam, a drug with muscarinic activity. (C) 1999 Elsevier Science Ltd. All rights reserved.

1998 - Telechelic oligomers by halogen atom transfer radical addition [Articolo su rivista]
Bellesia, Franco; Forti, Luca; E., Gallini; Ghelfi, Franco; Libertini, Emanuela; Pagnoni, Ugo Maria
abstract

Monodispersed telechelic oligomers have been efficiently prepared by Fe-0-FeCl3 promoted halogen atom transfer radical of functional telogens and taxogens, (C) 1998 Elsevier Science Ltd. All rights reserved.

1997 - Acetals by AlFe-pillared montmorillonite catalysis [Articolo su rivista]
Cramarossa, Maria Rita; Forti, Luca; Ghelfi, Franco
abstract

AlFe-pillared montmorillonite is an efficient catalyst for acetals preparation in CH2Cl2 at room temperature

1997 - Ferrocene promoted addition of methyl 2,2-dichloro-carboxylates to 1-alkenes [Articolo su rivista]
Forti, Luca; Ghelfi, Franco; Pagnoni, Ugo Maria
abstract

The Kharasch addition of methyl 2,2-dichlorocarboxylates to 1-alkenes is promoted by ferrocene in DMF at 100 degrees C, affording 1:1 adducts in satisfactory yields.

1997 - Halogen atom transfer radical addition of alpha-polychloroesters to olefins promoted by Fe-0 filings [Articolo su rivista]
Forti, Luca; Ghelfi, Franco; Libertini, Emanuela; Pagnoni, Ugo Maria; Soragni, Ercole
abstract

The Kharasch addition of methyl 2,2-dichlorocarboxylates or trichloro acetic acid derivatives to alkenes, affording the corresponding 1:1 adducts, is promoted by the iron filings/N,N-dimethylformamide system.

1997 - Halogen atom transfer radical cyclization of N-allyl-N-benzyl-2,2-dihaloamides to 2-pyrrolidinones, promoted by Fe-0-FeCl3 or CuCl-TMEDA [Articolo su rivista]
M., Benedetti; Forti, Luca; Ghelfi, Franco; Pagnoni, Ugo Maria; R., Ronzoni
abstract

The halogen atom transfer radical cyclization of a N-allyl-N-benzyl-2,2-dihaloamides to 2-pyrrolidinones has been carried out in high yields under mild conditions, in a reaction promoted by CuCl-TMEDA or Fe-0-FeCl3 in acetonitrile or N,N-dimethylformamide, respectively.

1997 - Reactivity of transition-metal-modified, Keggin-type heteropolycomplexes in the homogeneous oxidation of cyclohexane and adamantane [Articolo su rivista]
Cramarossa, Maria Rita; Forti, Luca; M. A., Fedotov; L. G., Detusheva; V. A., Likholobov; L. I., Kuznetsova; G. L., Semin; F., Cavani; F., Trifirò
abstract

The oxidation of cyclohexane and adamantane catalyzed by Keggin-type transition-metal-containing heteropolycomplexes (TMC-HPCs) is reported. The reactions were carried out at 20 degrees C in CCl4 or in CH2Cl2 as reaction solvent in the presence of t-butylhydroperoxide (t-BuOOH) as oxidizing agent. A radical chain mechanism of oxidation by t-BuOOH in the presence of molecular oxygen is proposed for all complexes tested, involving a R-H hydrogen atom abstraction by an intermediate active form of the TMC-HPC. Under the examined conditions molecular oxygen was necessary to initiate the reaction, The preparation of Co(II), Fe(III) and Cr(III)-HPC samples and their composition under reaction conditions are also discussed, based on P-31-NMR and W-VIS absorption spectra.

1997 - The Fe-0 promoted addition of CCl4 and CCl3Br to olefins [Articolo su rivista]
Bellesia, Franco; Forti, Luca; Ghelfi, Franco; Pagnoni, Ugo Maria
abstract

The radical addition of CCl4 or CCl3Br to olefins is efficiently promoted by iron filings in N,N-dimethylformamide under mild conditions.

1996 - Fe-0 initiated halogen atom transfer radical addition of methyl 2-Br-2-Cl-carboxylates to olefins [Articolo su rivista]
Forti, Luca; Ghelfi, Franco; Pagnoni, Ugo Maria
abstract

The halogen atom transfer radical addition (HATRA) of methyl 2-Br-2-Cl-carboxylates to alkenes is obtained in good yields by catalytic amounts of iron filings in dimethylformamide/1,2-dichloroethane at 80 degrees C under argon.

1996 - N-(2-hydroxyalkyl)-2,2-dihaloamides by amino-de-alkoxylation of methyl 2,2-dihalocarboxylates [Articolo su rivista]
Forti, Luca; Ghelfi, Franco; Grandi, Romano; Libertini, Emanuela; Pagnoni, Ugo Maria
abstract

The chemoselective N-acylation of 2-amino alcohols with methyl 2,2-dichloro carboxylates occurs smoothly and in excellent yields.

1996 - The CuBr/Fe-0 promoted radical addition of methyl 2-Br-2-Cl-carboxylates to olefins [Articolo su rivista]
Forti, Luca; Ghelfi, Franco; M., Lodi Lancellotti; Pagnoni, Ugo Maria
abstract

Methyl 2-Br-2-Cl-carboxylates afford 2-alkyl-2-Cl-4-Br-carboxylates in fair yields by reaction with terminal alkenes, under mild conditions, through a radical process promoted by CuBr/Fe-0 in DMF/CH2Cl2

1996 - Zinc promoted addition of methyl 2,2-dihalocarboxylates to carbonyl compounds [Articolo su rivista]
Benincasa, Marta; Forti, Luca; Ghelfi, Franco; Libertini, Emanuela; Pagnoni, Ugo Maria
abstract

Methyl 2,2-dihalocarboxylates add easily to carbonyl compounds in fair to good yields through the intermediate formation of 2-haloester enolates; the reaction is promoted by zinc, following a ''Barbier'' type procedure.

1995 - Amino-de-Alkoxylation of Methyl 2,2-Dihalocarboxylates [Articolo su rivista]
Forti, Luca; Ghelfi, Franco; F., Munari; Pagnoni, Ugo Maria
abstract

N-alkyl-2,2-dihaloamides were efficiently prepared by amino-de-alkoxylation of methyl 2,2-dihalocarboxylates, at room temperature without solvents and promoters. Excellent yields of anilides were obtained after addition of AlCl3.

1995 - Oxidation of nitrobenzene, chlorobenzenes and chlorophenols using liquid-phase ruthenium catalysts [Articolo su rivista]
A., Morvillo; Forti, Luca; M., Bressan
abstract

Ruthenium trichloride and various ruthenium(II) complexes, such as [Ru(H2O)(2) (L)(4)](BF4)(2) and [RuCl2(L)(4)] (L=DMSO), are effective catalysts for the complete oxidation of substituted aromatics at ambient temperature and in the liquid phase (double phase or water with suitable amounts of a surfactant agent) in the presence of an oxidizing agent, preferably monopersulfate. The oxidation of benzoic acid, chloro-, bromo-, iodo- and nitrobenzene and a number of polychlorobenzenes and polychlorophenols was followed by monitoring the nature and the relative amounts of the final products; chlorinated substrates are mainly converted into hydrochloric acid and carbon dioxide. Factors such as solvent, substrate, oxidant and concentration affect the reactions. The most favorable reaction conditions require the presence of polar media, like nitromethane or water. The fate of the catalyst during the various stages of the reactions was followed, which gave indications of the nature of the active species involved. The overall results imply that a strongly oxidizing ruthenium derivative is responsible for the effective oxidation with monopersulfate, tentatively a highly reactive peroxoruthenium species [Ru-O-O-SO3H]. Oxidation of chlorobenzene is first order in both the substrate and the catalysts and an n (or inverse) kinetic isotope effect is measured. The rates of the oxidation of mono-substituted benzenes depend upon the degree of deactivation of the aromatic ring, thus suggesting an initial electrophilic attack followed by a series of faster steps. Phenol and polychlorophenols are more sensitive to oxidation than substituted benzenes.

1995 - Reductive homo-coupling of methyl 2-Br-2-Cl-carboxylates promoted by CuBr-Fe0 [Articolo su rivista]
M., Benincasa; Forti, Luca; Ghelfi, Franco; Pagnoni, Ugo Maria
abstract

Dimethyl 2,3-dialkyl-2,3-dichloro-butanedioates are efficiently prepared in dimethylformamide or dimethylsulphoxide, through reductive homo-coupling of methyl 2-bromo-2-chlorocarboxylates promoted by CuBr-Fe-0.

1995 - Stereoselective Dehydrobromination of alkyl alpha-Br-alpha-Cl-carboxylates [Articolo su rivista]
Forti, Luca; Ghelfi, Franco; Pagnoni, Ugo Maria
abstract

(Z)-Alkyl alpha-Cl-alpha,beta-unsaturated esters are prepared in excellent yields by stereoselective dehydrobromination of alkyl alpha-Br-alpha-Cl-carboxylates with LiCl-Li2CO3 in dimethylformamide.

1995 - The CuBr/Fe promoted olefin alkylation of 2-Br-2-Cl-carboxylate methyl esters [Articolo su rivista]
Forti, Luca; Ghelfi, Franco; Pagnoni, Ugo Maria
abstract

Methyl 2-Br-2-Cl-carboxylates react with 1-octene in DMF/CH2Cl2 in a radical process promoted by CuBr/Fe, yielding 2-alkyl-2-Cl-4-Br-carboxylates smoothly and efficiently.

1994 - A Structural Study of a Novel MMO Model, [Fe2O(H2O)2(tris((1-methylimidazol-2-yl)methyl)amine)2]4+, and New Mechanistic Aspects of Alkane Functionalization Including Enhanced Catalytic Activity of the Aqua Complex [Abstract in Rivista]
R. H., Fish; R. M., Buchanan; S., Chen; J. F., Richardson; Bressan, Mario; Forti, Luca; A., Morvillo
abstract

A new biomimetic structural model of the active site of the MMO enzyme, [Fe2O(H2O)2(tris((1-methylimidazol-2-yl)methyl)amine)2]4+, has been characterized by single-crystal X-ray analysis and shown to functionalize cyclohexane, toluene, adamantane, propane, and ethane, in the presence of t-butyl hydroperoxide (TBHP) and O2 gas, with rates that are faster than the comparable mu-OAc bridged complex. This supports a mechanism of ligand replacement by the oxidant, TBHP, as rate -determining step in the formation of the active FeOFe=O oxidant. The use of a radical clock substrate, trans-2-phenylmethylcyclopropane, to support a free radical mechanism for the functionalization process, shows only ring-opened products, 1-phenyl-but-3-en-1-ol and a trace of its corresponding ketone. Thus, the initially formed cyclopropylcarbinyl radical rapidly rearranges to the 1-phenyl-but-3-enyl radical before O2 trapping to form a hydroperoxide.

1994 - An efficient procedure to alfa-hydroxyaldehyde dimethyl acetals [Articolo su rivista]
M., Boni; Forti, Luca; Ghelfi, Franco; Pagnoni, Ugo Maria
abstract

Alpha-Hydroxyaldehyde dimethyl acetals are prepared efficiently by conversion of alpha-haloaldehyde dimethyl acetals into alpha-haloaldehyde hemiacetal acetates and subsequent methanolysis promoted by lithium methoxide.

1994 - Biomimetic Oxidation Studies. 8. Structure of a New MMO Active Site Model, [Fe2O(H2O)2(tris((1-methylimidazol-2-yl)methyl)amine)2]4+, and Role of the Aqua Ligand in Alkane Functionalization Reactions [Articolo su rivista]
R. M., Buchanan; S., Chen; J. F., Richardson; M., Bressan; Forti, Luca; A., Morvillo; R. H., Fish
abstract

In this communication, we report on an X-ray crystallographicstudy of a new MMO active site analogue, [ Fe20(H20)2(tmima),]-(Cl04)4, 2, and on important mechanistic aspects of the C-Hbond functionalization reaction of several hydrocarbons with 2as the catalyst using anhydrous TBHP in the presence of oxygengas (02) and acetonitrile (CH3CN) as the solvent. We willdemonstrate that, by removing the p-OAc group and replacingit with terminal H20 ligands, complex 2 enhances the numberof turnovers/h for the alkane functionalization reaction, whileUV-vis and 1H NMR spectroscopic data suggest that 2 retains its integrity upon reaction with TBHP. In addition, we have further verified the previously found free radical mechanism for alkane functionalization with 2 by the use of a radical clock substrate, trans-2-phenylmethylcyclopropane to show only ring openedproducts that must emanate from a cyclopropylcarbinyl radical rearrangement.

1994 - Effective ruthenium-catalyzed oxidation of chlorinated olefins by monopersulfate in aqueous medium [Articolo su rivista]
M., Bressan; Forti, Luca; A., Morvillo
abstract

In aqueous medium, in the presence of a surfactant agent and of catalytic amounts of ruthenium derivatives, monopersulfate effectively oxidizes alpha-chlorinated olefins to hydrochloric acid and the appropriate carboxylic acid and/or carbon dioxide, with rates in excess of 100 turnover min(-1) at room temperature.

1993 - Iodosobenzene and persulfate oxygenation of saturated hydrocarbons catalyzed by diphosphino complexes of ruthenium(III) [Articolo su rivista]
M., Bressan; Forti, Luca; A., Morvillo
abstract

Hydroxylation and/or ketonization of alkanes (adamantane, cyclo-octene, hexane) is achieved by using iodosobenzene or aqueous mono-persulfate at room temperature in the presence of catalytic amounts of ruthenium(III)-diphosphino complexes dissolved in dichloromethane. On the basis of the kinetic behavior, different oxidizing metal species are proposed for the oxidations with mono-persulfate and with iodosobenzene.

1993 - Ruthenium(II)-catalyzed oxidation of alcohols by persulfate [Articolo su rivista]
M., Bressan; Forti, Luca; Ghelfi, Franco; A., Morvillo
abstract

Alcohols are oxidized to carbonylic compounds by aqueous persulfate in the presence of catalytic amounts (0.2%) of ruthenium trichloride and of a choice of ruthenium(II) complexes. Secondary alcohols are quantitatively converted to ketones, whereas primary alcohols can be converted to aldehydes with good selectivities. A kinetic investigation suggests a concerted mechanism for the bi-electronic oxygen transfer.

1993 - Ruthenium-catalyzed oxidation of alkylaromatics by monopersulfate with preferential oxidative fission of the benzene ring [Articolo su rivista]
M., Bressan; Forti, Luca; A., Morvillo
abstract

Ethylbenzene and a number of alkylbenzenes variously substituted at the alkyl chains, i.e. benzyl alcohol, benzaldehyde, benzoic acid, benzonitrile, benzyl chloride and benzyl bromide, undergo effective oxidative fission of the benzene ring to carbon dioxide and water when reacting at room temperature with aqueous persulfate in the presence of catalytic amounts of ruthenium derivatives.

1992 - A Study of Materials Suitable to Produce Bioceramics with Controlled Porosity for Prosthetic Implants Stabilized by Bone Tissue Ingrowth [Articolo su rivista]
Menabue, Ledi; Forti, Luca; Pellacani, Gian Carlo; M., Fabbri; A., Krajewski; A., Ravaglioli; G., Giunchi
abstract

Porous bioceramic pieces obtained by various methods were studied and here reported. The materials taken into consideration are alumina and particularly hydroxyapatite. Among the various materials that allow the formation of large pores (pore makers) with high porosity percentage, the following showedto be the most suitable: polyurethane, naphtalene and strontium carbonate. The obtained pores are communicating and their average diameter is significally high to succeed in hosting cells and blood microvessels inside.

1991 - Coordination behavior of N-protected amino acids. Structural and spectroscopic study of complexes of CoII, NiII and CuII with N-(4-aminobenzoyl)glicine [Articolo su rivista]
Forti, Luca; Menabue, Ledi; Saladini, Monica
abstract

The structural and spectroscopic properties of complexes of N-(4-aminobenzoyl)glycine (HL) with Co(II), Ni(II) and Cu(II) have been investigated. The complex [{CoL2(H2O)2}n].2nH2O crystallizes in the space group P1BAR, with a = 8.027(2), b = 8.031(2), c = 10.033(1) angstrom, alpha = 96.52(2)degrees, beta = 100.17(2)degrees, gamma = 119.19(2)degrees and Z = 1; the nickel(II) homologue is isomorphous. The complex [(CuL2)n].4nH2O crystallizes in the space group P2(1)/n, with a = 4.673(1), b = 20.476(1), c = 12.026(1) angstrom, beta = 97.38(2)degrees and Z = 2. The cobalt(II) co-ordination geometry is tetragonally distorted octahedral with four oxygens from two water molecules and two carboxylate groups in the basal plane and two amino nitrogens in the apical position. The copper(II) complex shows a strongly distorted tetragonal-octahedral geometry with two oxygens and two amino nitrogens of different amino acidate anions in the basal plane and two oxygens in the off-z-axis position. The ligand acts as bidentate bridging giving rise to a polymeric one-dimensional chain in the cobalt(II) complex and two-dimensional layers in the copper(II) one.

1991 - Coordinative capability of propane-1,3-diamine: spectroscopic and structural properties of a complex of formula [Cd(pnH)4Cl2]Cl4 [Articolo su rivista]
G., Ciani; N., Masciocchi; A., Sironi; S., Bruni; F., Cariati; A., Pozzi; Forti, Luca; Manfredini, Tiziano; Menabue, Ledi
abstract

The compound [Cd(pnH)4Cl2]Cl4 (pn = propane-1,3-diamine) has been prepared and studied by means of spectroscopic (IR and Raman) and structural investigations. It crystallizes in the tetragonal P4nc space group (No. 104), with two formula units in a cell of dimensions: a = b = 11.972(4), c = 9.129(4) angstrom. The structure consists of an ionic packing of chlorine anions and of discrete [Cd(pnH)4Cl2]4+ cations; the Cd atoms are octahedrally coordinated to four nitrogen atoms of four distinct monoprotonated diamines and to two chlorine atoms in trans position to each other, showing very loose Cd-Cl bonding interactions (2.680(3) and 2.884(2) angstrom). The protonated ends of the propane-1,3-diamines interact with the non-coordinated chlorine ions via hydrogen bonding. The Cd-N and Cd-X stretching vibrations are discussed and assigned in the light of the known structures.

1991 - Solid state behaviour of N-tosyl-DL-asparagine-Cu(II)-2,2'-bipyridine system [Articolo su rivista]
Forti, Luca; Saladini, Monica; Sola, Marco
abstract

The structural and spectroscopic properties of [Cu(bipy)(tsasnO)2].2H2O are investigated. The complex crystallizes in the space group P2(1)/n, with a = 12.502(2), b = 15.298(2), c = 18.901(8) angstrom, beta = 96.80(2)-degrees and Z = 4. The complex consists of discrete [Cu(bipy)(tsasnO)2] units and lattice water molecules. The Cu(II) ion is coordinated to two bipy nitrogens and two carboxylate oxygens from two aminoacidate anions in a square-planar geometry.

Università degli studi di Modena e Reggio Emilia

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