title: Functional characterization of a vanillin dehydrogenase in Corynebacterium glutamicum.
abstract: Vanillin dehydrogenase (VDH) is a crucial enzyme involved in the degradation of lignin-derived aromatic compounds. Herein, the VDH from Corynebacterium glutamicum was characterized. The relative molecular mass (Mr) determined by SDS-PAGE was ~51 kDa, whereas the apparent native Mr values revealed by gel filtration chromatography were 49.5, 92.3, 159.0 and 199.2 kDa, indicating the presence of dimeric, trimeric and tetrameric forms. Moreover, the enzyme showed its highest level of activity toward vanillin at pH 7.0 and 30°C, and interestingly, it could utilize NAD(+) and NADP(+) as coenzymes with similar efficiency and showed no obvious difference toward NAD(+) and NADP(+). In addition to vanillin, this enzyme exhibited catalytic activity toward a broad range of substrates, including p-hydroxybenzaldehyde, 3,4-dihydroxybenzaldehyde, o-phthaldialdehyde, cinnamaldehyde, syringaldehyde and benzaldehyde. Conserved catalytic residues or putative cofactor interactive sites were identified based on sequence alignment and comparison with previous studies, and the function of selected residues were verified by site-directed mutagenesis analysis. Finally, the vdh deletion mutant partially lost its ability to grow on vanillin, indicating the presence of alternative VDH(s) in Corynebacterium glutamicum. Taken together, this study contributes to understanding the VDH diversity from bacteria and the aromatic metabolism pathways in C. glutamicum.
authors.affiliation: State Key Laboratory of Crop Stress Biology for Arid Areas and College of Life Sciences, Northwest A&F University, Yangling, Shaanxi 712100, PR China.
authors.forename: Wei
authors.initials: W
authors.lastname: Ding
authors.affiliation: 1] State Key Laboratory of Crop Stress Biology for Arid Areas and College of Life Sciences, Northwest A&F University, Yangling, Shaanxi 712100, PR China [2] Biomass Energy Center for Arid and Semi-Arid Lands, Northwest A&F University, Yangling, Shaanxi 712100, PR China.
authors.forename: Meiru
authors.initials: M
authors.lastname: Si
authors.affiliation: State Key Laboratory of Crop Stress Biology for Arid Areas and College of Life Sciences, Northwest A&F University, Yangling, Shaanxi 712100, PR China.
authors.forename: Weipeng
authors.initials: W
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Crop Stress Biology for Arid Areas and College of Life Sciences, Northwest A&F University, Yangling, Shaanxi 712100, PR China.
authors.forename: Yaoling
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: 1] State Key Laboratory of Crop Stress Biology for Arid Areas and College of Life Sciences, Northwest A&F University, Yangling, Shaanxi 712100, PR China [2] Biomass Energy Center for Arid and Semi-Arid Lands, Northwest A&F University, Yangling, Shaanxi 712100, PR China.
authors.forename: Can
authors.initials: C
authors.lastname: Chen
authors.affiliation: 1] State Key Laboratory of Crop Stress Biology for Arid Areas and College of Life Sciences, Northwest A&F University, Yangling, Shaanxi 712100, PR China [2] Biomass Energy Center for Arid and Semi-Arid Lands, Northwest A&F University, Yangling, Shaanxi 712100, PR China.
authors.forename: Lei
authors.initials: L
authors.lastname: Zhang
authors.affiliation: College of Plant Protection, Northwest A&F University, Yangling, Shaanxi 712100, PR China.
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Lu
authors.affiliation: Biomass Energy Center for Arid and Semi-Arid Lands, Northwest A&F University, Yangling, Shaanxi 712100, PR China.
authors.forename: Shaolin
authors.initials: S
authors.lastname: Chen
authors.affiliation: 1] State Key Laboratory of Crop Stress Biology for Arid Areas and College of Life Sciences, Northwest A&F University, Yangling, Shaanxi 712100, PR China [2] Biomass Energy Center for Arid and Semi-Arid Lands, Northwest A&F University, Yangling, Shaanxi 712100, PR China.
authors.forename: Xihui
authors.initials: X
authors.lastname: Shen
mesh_terms: D000445:Aldehyde Oxidoreductases / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D000447:Aldehydes / Q000378:metabolism; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D001547:Benzaldehydes / Q000378:metabolism; D055162:Biocatalysis; D002850:Chromatography, Gel; D048230:Corynebacterium glutamicum / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016297:Mutagenesis, Site-Directed; D009243:NAD / Q000378:metabolism; D010802:Phylogeny; D055503:Protein Multimerization; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D016415:Sequence Alignment; D013379:Substrate Specificity
chemical_list: D000447:Aldehydes; D001426:Bacterial Proteins; D001547:Benzaldehydes; D011994:Recombinant Proteins; D009243:NAD; C100058:vanillin; D000445:Aldehyde Oxidoreductases; C105452:vanillin dehydrogenase