Fonctionnalisation enzymatique de chitosane par des composés phénoliques : évaluation des propriétés biologiques et physico-chimiques de ces nouveaux biopolymères

Oxidation of ferulic acid and its ester (ethyl ferulate) by Myceliophtora thermophila laccase has been studied in aqueous medium under mild experimental conditions (30°C and pH 7.5) as a green process to synthesize natural neo-molecules. Enzymatic oxidation led to colored and colorless intermediaries for ferulic acid and ethyl ferulate, respectively. Additionally, ethyl ferulate was oxidized faster than ferulic acid. This procedure has led to dimeric major products with MM = 443 g/mol and MM = 386 g/mol for ethyl ferulate and ferulic acid, respectively. New synthesized molecules demonstrated important antioxidants properties with weak antibacterial and cytotoxic properties. With insoluble chitosan particles in the reaction medium, laccase was protected from inhibition due to oxidation products and the polymerization degree of these products was checked. In addition, the oxidation products reacted with the free NH2 groups forming covalent bonds of Schiff base type (C=N) at C-2 region. The majority of the oxidation products grafted onto chitosan was of dimeric form. This procedure led to colored and colorless chitosans by ferulic acid and ethyl ferulate, respectively, with new properties due to grafting of phenolic compounds. These chitosan derivatives presented interesting functional properties such as antioxidant, physico-chemical (thermal stability) and biological (cell adhesion) as well as the preservation of antibacterial properties of native chitosanL'oxydation de l'acide férulique et de son ester (le férulate d'éthyle) par la laccase de Myceliophtora thermophila a été étudiée en milieu aqueux et dans des conditions expérimentales douces (30 °C et pH 7,5) afin de synthétiser de nouvelles molécules naturelles grâce à un procédé vert. L'oxydation enzymatique a permis l'obtention d'intermédiaires colorés pour l'acide férulique et incolores pour le férulate d'éthyle. En outre, le férulate d'éthyle a été plus rapidement totalement oxydé que l'acide férulique. De plus, cette procédure a abouti à des produits majoritairement dimériques avec MM = 443 g/mol et MM = 386 g/mol pour le férulate d'éthyle et l'acide férulique, respectivement. Les nouvelles molécules synthétisées ont présenté des propriétés antioxydantes importantes avec de faibles propriétés antibactériennes et cytotoxiques. En présence du chitosane insoluble dans le milieu réactionnel, la laccase a été protégée de l'inhibition liée aux produits d'oxydation et le degré de polymérisation de ces produits a été contrôlé. De plus, les produits d'oxydation ont réagi avec les groupements NH2 libres permettant la formation de liaisons covalentes de type base de Schiff (C=N) au niveau du C2 sur le chitosane. La majorité des produits d'oxydation greffés sur le chitosane était de forme dimérique. Cette procédure a permis d'obtenir du chitosane coloré avec l'acide férulique et incolore avec le férulate d'éthyle tout en présentant de nouvelles propriétés dues au greffage de composés phénoliques. Ces chitosanes dérivés ont présenté des propriétés fonctionnelles intéressantes telles que antioxydantes, physico-chimiques (stabilisation thermique) et biologiques (adhésion cellulaire) ainsi que la conservation des propriétés antibactériennes du chitosane nati

Enzymatic functionalization of chitosan by phenolic compounds : evaluation of biological and physico-chemical properties of these new biopolymers

L'oxydation de l'acide férulique et de son ester (le férulate d'éthyle) par la laccase de Myceliophtora thermophila a été étudiée en milieu aqueux et dans des conditions expérimentales douces (30 °C et pH 7,5) afin de synthétiser de nouvelles molécules naturelles grâce à un procédé vert. L'oxydation enzymatique a permis l'obtention d'intermédiaires colorés pour l'acide férulique et incolores pour le férulate d'éthyle. En outre, le férulate d'éthyle a été plus rapidement totalement oxydé que l'acide férulique. De plus, cette procédure a abouti à des produits majoritairement dimériques avec MM = 443 g/mol et MM = 386 g/mol pour le férulate d'éthyle et l'acide férulique, respectivement. Les nouvelles molécules synthétisées ont présenté des propriétés antioxydantes importantes avec de faibles propriétés antibactériennes et cytotoxiques. En présence du chitosane insoluble dans le milieu réactionnel, la laccase a été protégée de l'inhibition liée aux produits d'oxydation et le degré de polymérisation de ces produits a été contrôlé. De plus, les produits d'oxydation ont réagi avec les groupements NH2 libres permettant la formation de liaisons covalentes de type base de Schiff (C=N) au niveau du C2 sur le chitosane. La majorité des produits d'oxydation greffés sur le chitosane était de forme dimérique. Cette procédure a permis d'obtenir du chitosane coloré avec l'acide férulique et incolore avec le férulate d'éthyle tout en présentant de nouvelles propriétés dues au greffage de composés phénoliques. Ces chitosanes dérivés ont présenté des propriétés fonctionnelles intéressantes telles que antioxydantes, physico-chimiques (stabilisation thermique) et biologiques (adhésion cellulaire) ainsi que la conservation des propriétés antibactériennes du chitosane natifOxidation of ferulic acid and its ester (ethyl ferulate) by Myceliophtora thermophila laccase has been studied in aqueous medium under mild experimental conditions (30°C and pH 7.5) as a green process to synthesize natural neo-molecules. Enzymatic oxidation led to colored and colorless intermediaries for ferulic acid and ethyl ferulate, respectively. Additionally, ethyl ferulate was oxidized faster than ferulic acid. This procedure has led to dimeric major products with MM = 443 g/mol and MM = 386 g/mol for ethyl ferulate and ferulic acid, respectively. New synthesized molecules demonstrated important antioxidants properties with weak antibacterial and cytotoxic properties. With insoluble chitosan particles in the reaction medium, laccase was protected from inhibition due to oxidation products and the polymerization degree of these products was checked. In addition, the oxidation products reacted with the free NH2 groups forming covalent bonds of Schiff base type (C=N) at C-2 region. The majority of the oxidation products grafted onto chitosan was of dimeric form. This procedure led to colored and colorless chitosans by ferulic acid and ethyl ferulate, respectively, with new properties due to grafting of phenolic compounds. These chitosan derivatives presented interesting functional properties such as antioxidant, physico-chemical (thermal stability) and biological (cell adhesion) as well as the preservation of antibacterial properties of native chitosa

Fonctionnalisation enzymatique de chitosane par des composés phénoliques (évaluation des propriétés biologiques et physico-chimiques de ces nouveaux biopolymères)

L'oxydation de l'acide férulique et de son ester (le férulate d'éthyle) par la laccase de Myceliophtora thermophila a été étudiée en milieu aqueux et dans des conditions expérimentales douces (30 C et pH 7,5) afin de synthétiser de nouvelles molécules naturelles grâce à un procédé vert. L'oxydation enzymatique a permis l'obtention d'intermédiaires colorés pour l'acide férulique et incolores pour le férulate d'éthyle. En outre, le férulate d'éthyle a été plus rapidement totalement oxydé que l'acide férulique. De plus, cette procédure a abouti à des produits majoritairement dimériques avec MM = 443 g/mol et MM = 386 g/mol pour le férulate d'éthyle et l'acide férulique, respectivement. Les nouvelles molécules synthétisées ont présenté des propriétés antioxydantes importantes avec de faibles propriétés antibactériennes et cytotoxiques. En présence du chitosane insoluble dans le milieu réactionnel, la laccase a été protégée de l'inhibition liée aux produits d'oxydation et le degré de polymérisation de ces produits a été contrôlé. De plus, les produits d'oxydation ont réagi avec les groupements NH2 libres permettant la formation de liaisons covalentes de type base de Schiff (C=N) au niveau du C2 sur le chitosane. La majorité des produits d'oxydation greffés sur le chitosane était de forme dimérique. Cette procédure a permis d'obtenir du chitosane coloré avec l'acide férulique et incolore avec le férulate d'éthyle tout en présentant de nouvelles propriétés dues au greffage de composés phénoliques. Ces chitosanes dérivés ont présenté des propriétés fonctionnelles intéressantes telles que antioxydantes, physico-chimiques (stabilisation thermique) et biologiques (adhésion cellulaire) ainsi que la conservation des propriétés antibactériennes du chitosane natifOxidation of ferulic acid and its ester (ethyl ferulate) by Myceliophtora thermophila laccase has been studied in aqueous medium under mild experimental conditions (30C and pH 7.5) as a green process to synthesize natural neo-molecules. Enzymatic oxidation led to colored and colorless intermediaries for ferulic acid and ethyl ferulate, respectively. Additionally, ethyl ferulate was oxidized faster than ferulic acid. This procedure has led to dimeric major products with MM = 443 g/mol and MM = 386 g/mol for ethyl ferulate and ferulic acid, respectively. New synthesized molecules demonstrated important antioxidants properties with weak antibacterial and cytotoxic properties. With insoluble chitosan particles in the reaction medium, laccase was protected from inhibition due to oxidation products and the polymerization degree of these products was checked. In addition, the oxidation products reacted with the free NH2 groups forming covalent bonds of Schiff base type (C=N) at C-2 region. The majority of the oxidation products grafted onto chitosan was of dimeric form. This procedure led to colored and colorless chitosans by ferulic acid and ethyl ferulate, respectively, with new properties due to grafting of phenolic compounds. These chitosan derivatives presented interesting functional properties such as antioxidant, physico-chemical (thermal stability) and biological (cell adhesion) as well as the preservation of antibacterial properties of native chitosanMETZ-SCD (574632105) / SudocNANCY1-Bib. numérique (543959902) / SudocNANCY2-Bibliotheque electronique (543959901) / SudocNANCY-INPL-Bib. électronique (545479901) / SudocSudocFranceF

Lipase catalyzed esterification of formic acid in solvent and solvent-free systems

International audienceEsterification reaction between formic acid and alcohols (C4, C8) catalyzed by lipases was performed in solvent and solvent-free systems. High Performance Liquid Chromatography (HPLC) was used to study the kinetic parameters (Michaelis-Menten) and the reaction conditions by monitoring the ester synthesis. The optimal conditions for formate ester synthesis were found to be: 0.5 M of formic acid, 1.5 M of alcohol, 2% of Novozym 435 at 40 degrees C and 400 rpm in acetonitrile solvent without molecular sieves. In solvent-free system, the optimal conditions were found to be: 1 M of formic acid, 10 M of alcohol, 2% of Novozym 435 at 40 degrees C and 400 rpm without molecular sieves. These optimal conditions resulted in maximum ester yield about 90% for 8 h in solvent system and 5 h in solvent-free system. Among all esterification reactions, the esterification reaction of formic acid with octan-1-ol in solvent-free system was the better regarding the quantity of esterified formic acid, catalytic efficiency of lipase and the required reaction time. Under these optimal conditions, the biocatalyst could be reused for six reaction cycles keeping around 97% of its initial activity after treatment process with n-hexane. Consequently, this enzymatic procedure in solvent-free system could provide formate esters for food and cosmetic applications

Enzymatic Oxidation of Ferulic Acid as a Way of Preparing New Derivatives

The ferulic acid (FA)-oxidation by Myceliophthora thermophila laccase was performed in phosphate buffer at 30 °C and pH 7.5 as an eco-friendly procedure. LC-MS analysis showed that oxidation products were four dehydrodimers (P1, P2, P3, P5) at MM = 386 g/mol, two dehydrotetramers (P6, P7) at MM = 770 g/mol and one decarboxylated dehydrodimer (P4) at MM = 340 g/mol. Structural characterization showed that FA-dehydrodimers were symmetric for P1 and P5 while asymmetric for P2, P3 and P4. Physicochemical characterization showed that oxidation products presented a higher lipophilicity than that of FA. Moreover, symmetric dimers and tetra dimers had a higher melting point compared to FA and its asymmetric dimers. Antioxidant and anti-proliferative assessments indicated that enzymatic oligomerization increased antioxidant and anti-proliferative properties of oxidation products for P2, P3 and P6 compared to FA. Finally, this enzymatic process in water could produce new molecules, having good antiradical and anti-proliferative activities

Physicochemical characterization of pectin grafted with exogenous phenols

International audiencePectin is a natural polysaccharide, having valuable properties that enable its use in many industrial fields. The aim of this work was to study the impact of pectin modification with phenols, on the properties of this biopolymer. Results suggested that the enzymatic grafting of ferulic acid (FA) oxidation products onto the pectin altered its morphological surface and its thermal properties. Moreover, modified pectin showed a less hygroscopic behavior when water activity is less than 0.50 and a higher ability to bound water above 0.5. Additionally, modified pectin became less viscous than the native pectin and presented different calciumdependent gelation behavior. Finally, a significant improvement of the antioxidant properties of pectin after functionalization was observed. As a conclusion, the modification of pectin with phenolic compounds appeared as a promising way to produce a polysaccharide with new properties that could enlarge the field of its potential applications

Laccase-catalysed oxidation of ferulic acid and ethyl ferulate in aqueous medium: A green procedure for the synthesis of new compounds

International audienceThe enzymatic oxidation of ferulic acid (FA) and ethyl ferulate (EF) with Myceliophthora thermophila laccase, as biocatalyst, was performed in aqueous medium using an eco-friendly procedure to synthesize new active molecules. First, the commercial laccase was ultrafiltrated allowing for the elimination of phenolic contaminants and increasing the specific activity by a factor of 2. Then, kinetic parameters of this laccase were determined for both substrates (FA, EF), indicating a higher substrate affinity for ethyl ferulate. Additionally, enzymatic oxidation led to the synthesis of a FA-major product, exhibiting a molecular mass of 386 g/mol and a EF-major product with a molecular mass of 442 g/mol. Structural analyses by mass spectrometry allowed the identification of dimeric derivatives. The optical properties of the oxidation products showed the increase of red and yellow colours, with FA-products compared to EF-products. Additionally, enzymatic oxidation led to a decrease of antioxidant and cytotoxic activities compared to initial substrates. Consequently, this enzymatic procedure in aqueous medium could provide new compounds presenting optical, antioxidant and cytotoxic interest

Reliable bioassays to detect potential hazard of paperboard food contact material extracts

Food contact materials (FCM) represent a major economic issue and a large field of innovation. Food packaging production must be in compliance with the European Regulation 1935/2004: the third article specifies that FCM must not transfer their constituents to food in quantities which could endanger human health under normal or foreseeable conditions of use. Indeed, these materials are not inert and, in addition to started substances, Non-Intentionally Added Substances (NIAS) are able to be released into the food. NIAS can be contaminants from recycled materials, impurities, synthetic residues, new substances formed along the packaging production chain… Then, NIAS could represent a larget part of migrating substances which are not risk assessed, some being unpredictabled (Grob et al., 2006 ; Skejevrak et al., 2005). The European Regulation 10/2011 on plastic materials, multilayers and articles intended to come into contact with food regulates NIAS. It requires that the notion of the risk engendered by a substance concerns the substance itself, the impurities of this substance and any reaction or degradation products that are foreseeable in the context of the intended use. However there are currently no available guidelines to assess the risk of NIAS. They may also be present in other FCM than plastics, such as paper and boards... Because of the complex nature of FCM extract, it is pertinent to evaluate the potential toxicity of FCM as a whole taking into account any « cocktail effect ». The objectives of this study are to provide to packaging and food companies scientific relevant tools combining physicochemical and toxicological strategies based on bioassays applied to FCM extracts, especially paper and boards. Four toxicological endpoints were checked as relevant for low dose exposure: cytotoxicity, genotoxicity , oxidative stress and endocrine disruption activities

Laccase-catalysed functionalisation of chitosan by ferulic acid and ethyl ferulate: Evaluation of physicochemical and biofunctional properties

International audienceChitosan and its derivatives functionalized by laccase-catalyzed oxidation of ferulic acid (FA) and ethyl ferulate (EF) were characterised for their physico-chemical, antioxidant and antibacterial properties. The enzymatic grafting of oxidised phenols led to FA-coloured and EF-colourless chitosan derivatives with good stability of colour and grafted phenols towards the chemical treatment by organic solvents. The efficiency of FA-products grafting onto chitosan was higher than that of EF-products. Moreover, the enzymatic grafting of phenols onto chitosan changed its morphological surface, increased its molecular weight and its viscosity. Furthermore, the chitosan derivatives presented improved antioxidant properties especially for FA-chitosan derivative when compared with chitosan with good antioxidant stability towards thermal treatment (100 degrees C/1 h). Chitosan and its derivatives showed also similar antibacterial activities and more precisely bactericidal activities. This enzymatic procedure provided chitosan derivatives with improved properties such as antioxidant activity, thermal antioxidant stability as well as the preservation of initial antibacterial activity of chitosan

Bioprocesses for the Biodiesel Production from Waste Oils and Valorization of Glycerol

The environmental context causes the use of renewable energy to increase, with the aim of finding alternatives to fossil-based products such as fuels. Biodiesel, an alternative to diesel, is now a well-developed solution, and its production from renewable resources makes it perfectly suitable in the environmental context. In addition, it is biodegradable, non-toxic and has low greenhouse gas emissions: reduced about 85% compared to diesel. However, the feedstock used to produce biodiesel competes with agriculture and the application of chemical reactions is not advantageous with a “green” process. Therefore, this review focuses only on bioprocesses currently taking an important place in the production of biodiesel and allow high yields, above 90%, and with very few produced impurities. In addition, the use of waste oils as feedstock, which now accounts for 10% of feedstocks used in the production of biodiesel, avoids competition with agriculture. To present a complete life-cycle of oils in this review, a second part will focus on the valorization of the biodiesel by-product, glycerol. About 10% of glycerol is generated during the production of biodiesel, so it should be recovered to high value-added products, always based on bioprocesses. This review will also present existing techniques to extract and purify glycerol. In the end, from the collection of feedstocks to the production of CO2 during the combustion of biodiesel, this review presents the steps using the “greener” possible processes