Cloned (Comment) | Organism |
---|---|
genes vanAB, functional expression in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain DH5alpha, VanA is expressed as a soluble protein, whereas VanB is expressed in inclusion bodies | Streptomyces sp. NL15-2K |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-hydroxy-3-methylbenzoate + O2 + NADH + H+ | Streptomyces sp. NL15-2K | - |
? | - |
? | |
m-anisate + O2 + NADH + H+ | Streptomyces sp. NL15-2K | - |
m-hydroxybenzoate + NAD+ + H2O + formaldehyde | - |
? | |
syringate + NADH + H+ + O2 | Streptomyces sp. NL15-2K | - |
? | - |
? | |
vanillate + O2 + NADH + H+ | Streptomyces sp. NL15-2K | i.e. protocatechiuc acid | 3,4-dihydroxybenzoate + NAD+ + H2O + formaldehyde | - |
? | |
veratrate + NADH + H+ + O2 | Streptomyces sp. NL15-2K | - |
isovanillate + NAD+ + H2O + formaldehyde | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Streptomyces sp. NL15-2K | Q08KC3 AND Q08KC3 | vanillate O-demethylase oxygenase subunits A and B | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-hydroxy-3-methylbenzoate + O2 + NADH + H+ | - |
Streptomyces sp. NL15-2K | ? | - |
? | |
4-hydroxy-3-methylbenzoate + O2 + NADH + H+ | - |
Streptomyces sp. NL15-2K | 4-hydroxy-3-(hydroxymethyl)benzoate + NAD+ + H2O + formaldehyde | - |
? | |
m-anisate + O2 + NADH + H+ | - |
Streptomyces sp. NL15-2K | m-hydroxybenzoate + NAD+ + H2O + formaldehyde | - |
? | |
additional information | substrate specificity for vanillate analogues by an in vivo assay using recombinant whole cells: among aromatic methyl ethers, vanillate, syringate, m-anisate, and veratrate are good substrates, whereas ferulate, vanillin, and guaiacol are not recognized by Streptomyces vanillate demethylase. Besides vanillate, 4-hydroxy-3-methylbenzoate is a better substrate than m-anisate and veratrate, and the 3-methyl group is efficiently oxidized to a hydroxymethyl group. The combination of a carboxyl group on the benzene ring and a hydroxyl group in the para-position relative to the carboxyl group may be preferable for substrate recognition by the enzyme. Demethylation by Streptomyces vanillate demethylase occurs only at the 3-position of veratrate and not at the 4-position | Streptomyces sp. NL15-2K | ? | - |
? | |
syringate + NADH + H+ + O2 | - |
Streptomyces sp. NL15-2K | ? | - |
? | |
syringate + NADH + H+ + O2 | 3,4-dihydroxy-5-methoxybenzoate, and gallate are produced from syringate, one of the two methoxy groups in the meta-position of syringate is demethylated first, and then the second methoxy group is demethylated at a slower rate | Streptomyces sp. NL15-2K | 3,4-dihydroxy-5-methoxybenzoate + gallate + NAD+ + H2O + formaldehyde | - |
? | |
vanillate + O2 + NADH + H+ | i.e. protocatechiuc acid | Streptomyces sp. NL15-2K | 3,4-dihydroxybenzoate + NAD+ + H2O + formaldehyde | - |
? | |
veratrate + NADH + H+ + O2 | - |
Streptomyces sp. NL15-2K | isovanillate + NAD+ + H2O + formaldehyde | - |
? | |
veratrate + NADH + H+ + O2 | demethylation product of veratrate is isovanillate rather than vanillate, 1H-NMR analysis. O-Demethylation of veratrate by Streptomyces vanillate demethylase occurs only at the meta-position relative to the carboxyl group | Streptomyces sp. NL15-2K | isovanillate + NAD+ + H2O + formaldehyde | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | 1 x 40000, subunit A + 1 * 33000, subunit B, recombinant enzyme, SDS-PAGE | Streptomyces sp. NL15-2K |
Synonyms | Comment | Organism |
---|---|---|
VanA | - |
Streptomyces sp. NL15-2K |
vanAB | - |
Streptomyces sp. NL15-2K |
VanB | - |
Streptomyces sp. NL15-2K |
vanillate demethylase | - |
Streptomyces sp. NL15-2K |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADH | - |
Streptomyces sp. NL15-2K |