Identification and characterization of the vanillin dehydrogenase YfmT in Bacillus subtilis 3NA

Abstract

With vanillin as one of the most important flavoring agents, many efforts have been made to optimize its biotechnological production from natural abundant substrates. However, its toxicity against the hosts results in rather low yields and product concentrations. Bacillus subtilis as a soil-dwelling bacterium is a possible lignin-derived compound-degrading microorganism. Therefore, its vanillin and ferulic acid metabolism was investigated. With a rather high tolerance for vanillin up to 20 mM, it is a promising candidate to produce natural vanillin. In this study, the well-studied phenolic acid decarboxylases PadC and BsdBCD could be ascribed to function as the only enzymes in B. subtilis 3NA converting ferulic acid to 4-vinylguaiacol and vanillic acid to guaiacol, respectively. As vanillin also becomes converted to guaiacol, a previous conversion to vanillic acid was assumed. Usage of bioinformatic tools revealed YfmT, which could be shown to function as the only vanillin dehydrogenase in B. subtilis 3NA. Thus, YfmT was further characterized regarding its temperature and pH optima as well as its substrate range. Vanillin and ferulic acid metabolic routes in the tested B. subtilis strain were revealed, a direct conversion of ferulic acid to vanillin, however, could not be found.

Comparative Genomics of Bacillus amyloliquefaciens Strains Reveals a Core Genome with Traits for Habitat Adaptation and a Secondary Metabolites Rich Accessory Genome

“…Both phenolic acid decarboxylases padC and bsdBCD ( yclBCD ) of B. subtilis were mined. The putative LysR-type regulator encoded by bsdA ( yclA ) gene upstream of the bsdBCD operon revealed is the transcriptional activator of bsdBCD expression in response to phenolic acids were also mined (Graf et al, 2016 ). Dibenzothiophene (DBT) is the model compound for this class of molecules.…”

Evaluation of process involved in the production of aromatic compounds in Gram‐negative bacteria isolated from vanilla ( Vanilla planifolia ex. Andrews) beans

“…Some Gram‐negative and Gram‐positive bacteria can catalyse FA bioconversion to intermediate compounds such as vanillic alcohol, VA and vanillin (Vyrides et al ; Graf et al ; Mishra et al ; Paz et al ). The AC produced from FA have been studied in different strains of Pseudomonas sp.…”

“…Enterobacter sp. Px6-4, a reference strain, isolated from V. planifolia roots produced decarboxylase activity (0Á5 U ml À1 ) and was capable of degrading FA at maximum growth (15 h) (Li et al (Vyrides et al 2015;Graf et al 2016;Mishra et al 2016;Paz et al 2016). The AC produced from FA have been studied in different strains of Pseudomonas sp.…”

Cross Talk among Transporters of the Phosphoenolpyruvate-Dependent Phosphotransferase System in Bacillus subtilis

“…Unless otherwise specified, the start and stop codons of the target genes were fused in order to prevent the polar effect in all deletion strategies. In the first strategy, desired genes were deleted using temperature-sensitive derivatives of the pMW521.1 plasmid ( 66 ). pMW521.1 is an integration shuttle vector consisting of ori pUC18 for E. coli and temperature-sensitive ori pE194 for B. subtilis .…”