Laccase mediated-synthesis of hydroxycinnamoyl-peptide from ferulic acid and carnosine

• Published in: Journal of biotechnology 2016-06, Vol.227, p.83-93
• Main Authors: Aljawish, Abdulhadi, Chevalot, Isabelle, Madad, Nidal, Paris, Cédric, Muniglia, Lionel
• Format: Article
• Language: English
• Subjects: Antioxidant activity; Antioxidants; Benzothiazoles - metabolism; Biocatalysis - drug effects; Biotechnology; Caco-2 Cells; Caco2 cells; Carnosine; Carnosine - metabolism; Carnosine - pharmacology; Cell Proliferation - drug effects; Chemical and Process Engineering; Chromatography, High Pressure Liquid; Coumaric Acids - metabolism; Coumaric Acids - pharmacology; Coupling; Derivatives; Engineering Sciences; Ferulic acid; Humans; Inhibitory Concentration 50; Kinetics; Laccase; Laccase - metabolism; Mass Spectrometry; Molecular structure; Optimization; Oxidation-Reduction - drug effects; Peptides - metabolism; Solubility; Spectrophotometry, Ultraviolet; Sulfonic Acids - metabolism; Time Factors
• ISSN: 0168-1656; 1873-4863
• DOI: 10.1016/j.jbiotec.2016.04.021

•Enzymatic synthesis of carnosine derivatives with ferulic acid in aqueous medium under mild conditions.•The enzymatic process created two carnosine derivatives exhibiting the same molecular mass (760g/mol) but with different chemical structures.•Structural analyses showed the presence of one carnosine molecule and three ferulic acid molecules in each derivative.•Enzymatic functionalization improved both antioxidant and anti-proliferative properties of carnosine derivatives in comparison with carnosine. Carnosine (CAR) dipeptide was functionalized with ferulic acid (FA) as substrate using laccase from Myceliophtora thermophila as biocatalyst. The enzymatic reaction was performed in aqueous medium under mild conditions (pH 7.5, 30°C) as an eco-friendly procedure. Results showed that this enzymatic process led to the synthesis of two new derivatives (P1, P2), from the coupling between CAR and FA derived products. Conditions allowing a high production of P1, P2 derivatives were determined with an optimal ratio of (FA: CAR) of (1:1.6) at optimal time reaction of 8h. Under these optimal conditions, the coupling between CAR and FA-products was demonstrated, resulting in the decrease of −NH2 groups (almost 50%) as quantified via derivatization. Due to the presence of FA in the structure of these new derivatives, they exhibited higher hydrophobic property than carnosine. Structural analyses by mass spectrometry showed that P1 and P2 (FA-CAR) derivatives exhibited the same molecular mass (MM 770g/mol) containing one CAR-molecule and three FA-molecules but with different chemical structures. Furthermore, these derivatives presented improved antioxidant (almost 10 times) and anti-proliferative (almost 18 times) properties in comparison with CAR. Moreover, P1 derivative exhibited higher antioxidant and anti-proliferative activities than P2 derivative, which confirmed the different structures of P1 and P2. These results suggested that the oxidized phenols coupling with carnosine is a promising process to enhance the CAR-properties.