Generation of 4-vinylguaiacol through a novel high-affinity ferulic acid decarboxylase to obtain smoke flavours without carcinogenic contaminants

• Main Authors: Berger, R.G (Author), Detering, T. (Author), Mundry, K. (Author)
• Format: Article
• Language: English
• Published: Public Library of Science 2021
• Subjects: 4 vinylguaiacol; 4-vinylguaiacol; affinity chromatography; amino acid sequence; amino terminal sequence; Article; beta sheet; budding yeast; caffeic acid; carboxylesterase; Carboxylic Ester Hydrolases; carboxylyase; Carboxy-Lyases; carcinogen; Carcinogens; chemistry; cinnamic acid; complementary DNA; controlled study; Cordyceps; coumaric acid; Coumaric Acids; decarboxylation; enzyme active site; enzyme activity; enzyme phosphorylation; enzyme purification; enzyme stability; FAD gene; ferulic acid; ferulic acid decarboxylase; flavor; flavoring agent; Flavoring Agents; gene; gene amplification; gene expression; green chemistry; guaiacol; Guaiacol; hydrogen bond; hydrophilicity; isolation and purification; Komagataella; Komagataella phaffii; metabolism; Michaelis Menten kinetics; molecular cloning; nickel complex; nonhuman; phenylacrylic acid decarboxylase; RNA isolation; Saccharomycetales; Sanger sequencing; Schiff base; Schizophyllum; Schizophyllum commune; sequence alignment; sinapic acid; smoke; synthesis; unclassified drug
• Online Access: View Fulltext in Publisher

 Traditional smoke flavours bear the risk of containing a multitude of contaminating carcinogenic side-products. Enzymatic decarboxylation of ferulic acid released from agro-industrial side-streams by ferulic acid esterases (FAE) enables the sustainable generation of pure, food grade 4-vinylguaiacol (4-VG), the impact compound of smoke flavour. The first basidiomycetous ferulic acid decarboxylase (FAD) was isolated from Schizophyllum commune (ScoFAD) and heterologously produced by Komagataella phaffii. It showed a molecular mass of 21 kDa, catalytic optima at pH 5.5 and 35 C, and a sequence identity of 63.6% to its next relative, a FAD from the ascomycete Cordyceps farinosa. The ScoFAD exhibited a high affinity to its only known substrate ferulic acid (FA) of 0.16 mmol L-1 and a turnover number of 750 s-1. The resulting catalytic efficiency kcat KM-1 of 4,779 L s-1 mmol-1 exceeded the next best known enzyme by more than a factor of 50. Immobilised on AminoLink Plus Agarose, ScoFAD maintained its activity for several days. The combination with FAEs and agro-industrial side-streams paves the way for a new generation of sustainable, clean, and safe smoke flavours. © 2020 Detering et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.