Biotransformation of ferulic acid to protocatechuic acid by Corynebacterium glutamicum ATCC 21420 engineered to express vanillate O-demethylase

Abstract

Ferulic acid (4-hydroxy-3-methoxycinnamic acid, FA) is a lignin-derived phenolic compound abundant in plant biomass. The utilization of FA and its conversion to valuable compounds is desired. Protocatechuic acid (3,4-dihydroxybenzoic acid, PCA) is a precursor of polymers and plastics and a constituent of food. A microbial conversion system to produce PCA from FA was developed in this study using a PCA-producing strain of Corynebacterium glutamicum F (ATCC 21420). C. glutamicum strain F grown at 30 °C for 48 h utilized 2 mM each of FA and vanillic acid (4-hydroxy-3-methoxybenzoic acid, VA) to produce PCA, which was secreted into the medium. FA may be catabolized by C. glutamicum through proposed (I) non-β-oxidative, CoA-dependent or (II) β-oxidative, CoA-dependent phenylpropanoid pathways. The conversion of VA to PCA is the last step in each pathway. Therefore, the vanillate O-demethylase gene (vanAB) from Corynebacterium efficiens NBRC 100395 was expressed in C. glutamicum F (designated strain FVan) cultured at 30 °C in AF medium containing FA. Strain C. glutamicum FVan converted 4.57 ± 0.07 mM of FA into 2.87 ± 0.01 mM PCA after 48 h with yields of 62.8% (mol/mol), and 6.91 mM (1064 mg/L) of PCA was produced from 16.0 mM of FA after 12 h of fed-batch biotransformation. Genomic analysis of C. glutamicum ATCC 21420 revealed that the PCA-utilization genes (pca cluster) were conserved in strain ATCC 21420 and that mutations were present in the PCA importer gene pcaK.Electronic supplementary materialThe online version of this article (doi:10.1186/s13568-017-0427-9) contains supplementary material, which is available to authorized users.

Investigating the Efficiency of Hydroxycinnamic Acids to Inhibit the Production of Enniatins by Fusarium avenaceum and Modulate the Expression of Enniatins Biosynthetic Genes

“…According to our results, neither vanillin nor methoxyquinone were detected in FER-supplemented media inoculated by F. avenaceum. We however detected the presence of PROTO which could be explained by the metabolisation of VAN catalyzed by a vanillate-O-demethylase [39,40]. In our experiments, while more than 4 µmoles of FER have disappeared from the culture broths after 7 days of incubation at pH 4 or 10 days at pH 7, the sum of quantified VAN + PROTO acids did not exceed 1 µmole.…”

Advances and Prospects of Phenolic Acids Production, Biorefinery and Analysis

“…Hypogallic acid can be produced in E. coli when chorismate is first converted to isochorismate, then to 2,3-dihydroxy-2,3-dihydrobenzoic acid (2,3-dihydro-2,3-DHBA), and finally to this phenolic acid by the mediation of isochorismate synthase, isochorismatase, and 2,3-dihydro-2,3-DHBA dehydrogenase, respectively [ 221 ]. Protocatechuic acid is synthesised from ferulic acid through non-β-oxidative or β-oxidative CoA-dependent pathways, where the last step is the biotransformation of vanillic acid mediated by vanillate O-demethylase [ 222 , 223 ]. Vanillic acid is synthesised from ferulic after disruption of vanA and vanB genes from Pseudomonas fluorescens and P. putida using p -coumaric acid or ferulic acid extracted from corn bran [ 205 , 224 ].…”

A Timed Off-Switch for Dynamic Control of Gene Expression in Corynebacterium Glutamicum

“…Fcs, feruloyl-CoA synthetase; Ech, enoyl-CoA hydratase/aldolase; Vdh, vanillin dehydrogenase; VanAB, A and B subunits of vanillate demethylase encoded by vanA and vanB; TCA, tricarboxylic acid cycle; VanR, PadR-type repressor encoded by vanR; vanK, gene coding for vanillate transport protein VanK; cg2464 + cg2465, genes coding for hypothetical proteins; aceE, aceF, and lpd, genes coding for E1-, E2-and E3-subunits of pyruvate dehydrogenase complex (PDHC), respectively; P vanABK , promotor region of vanABK and its −10-region-optimized derivative indicated by (*). This figure was adapted and compiled with information from diverse references (Kalinowski et al, 2003;Merkens et al, 2005;Brinkrolf et al, 2006;Chaudhry et al, 2007;Shen et al, 2012;Pfeifer-Sancar et al, 2013;Eikmanns and Blombach, 2014;Ding et al, 2015;Morabbi Heravi et al, 2015;Kallscheuer et al, 2016;Okai et al, 2017). Menkel et al, 1989;Cordes et al, 1992 pJC4-ilvBNCE pJC4 with coding sequence for native ilvBNCE genes from C. glutamicum WT Radmacher et al, 2002 was prepared by adding 18 g agar-agar L −1 to liquid medium in 9-cm Petri dishes, which were incubated at the abovementioned temperature after inoculation.…”

“…Corynebacterium glutamicum is able to grow on the lignin-derived phenolic compounds ferulic acid (FA), vanillin (Van), and vanillic acid (VA) and, compared with other microbial systems, is less susceptible to growth inhibition by these molecules (Shen et al, 2012 ; Ding et al, 2015 ; Becker and Wittmann, 2019 ). The degradation of Van and VA proceeds via the vanillate pathway yielding protocatechuic acid (PCA), which is subsequently converted by the β-ketoadipate pathway to acetyl-CoA and succinyl-CoA entering eventually the tricarboxylic acid (TCA) cycle [ Figure 1 (Merkens et al, 2005 ; Brinkrolf et al, 2006 ; Shen et al, 2012 ; Kallscheuer et al, 2016 ; Okai et al, 2017 )]. Within the vanillate pathway, FA might be converted in a cascade via Van and VA to PCA (Merkens et al, 2005 ; Brinkrolf et al, 2006 ; Shen et al, 2012 ).…”