Cloned (Comment) | Organism |
---|---|
overexpression of wild-type and mutant enzymes | Escherichia coli |
Crystallization (Comment) | Organism |
---|---|
purified recombinant mutant enzymes Y17F, Y144F, D147N, R253A, and R253K, hanging drop method, 20°C, 10 mg/ml protein in solution mixed with equal volume of well solution containing 26-28% PEG 4000, 9-12% methylpentanediol, 100 mM HEPES, pH 7.5, microseeding, 2 days, X-ray diffraction structure determination and analysis at 1.7-2.4 A resolution, modeling | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
D147N | site-directed mutagenesis, mutation of an active site residue, inactive mutant | Escherichia coli |
R253A | site-directed mutagenesis, altered kinetics, highly increased Km for S-adenosyl-L-methionine compared to the wild-type enzyme | Escherichia coli |
R253K | site-directed mutagenesis, altered kinetics compared to the wild-type enzyme | Escherichia coli |
R253M | site-directed mutagenesis, altered kinetics compared to the wild-type enzyme | Escherichia coli |
R253Q | site-directed mutagenesis, altered kinetics compared to the wild-type enzyme | Escherichia coli |
Y144F | site-directed mutagenesis, mutation of an active site residue, highly increased Km for S-adenosyl-L-methionine, highly reduced activity compared to the wild-type enzyme | Escherichia coli |
Y17F | site-directed mutagenesis, mutation of an active site residue, highly reduced activity compared to the wild-type enzyme | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | single turnover reactions, stopped-flow, wild-type and mutant enzymes | Escherichia coli | |
0.0006 | - |
8-Amino-7-oxononanoate | pH 9.0, recombinant mutant R253M | Escherichia coli | |
0.0008 | - |
8-Amino-7-oxononanoate | pH 9.0, recombinant mutant R253K | Escherichia coli | |
0.001 | - |
8-Amino-7-oxononanoate | pH 9.0, recombinant wild-type enzyme | Escherichia coli | |
0.0014 | - |
8-Amino-7-oxononanoate | pH 9.0, recombinant mutant R253Q | Escherichia coli | |
0.0022 | - |
8-Amino-7-oxononanoate | pH 9.0, recombinant mutant R253A | Escherichia coli | |
0.01 | - |
S-adenosyl-L-methionine | pH 9.0, recombinant mutant R253K | Escherichia coli | |
0.011 | - |
S-adenosyl-L-methionine | pH 9.0, recombinant mutant R253Q | Escherichia coli | |
0.02 | - |
8-Amino-7-oxononanoate | pH 9.0, recombinant mutant Y17F | Escherichia coli | |
0.063 | - |
S-adenosyl-L-methionine | pH 9.0, recombinant mutant Y17F | Escherichia coli | |
0.1 | - |
S-adenosyl-(5')-3-methylthiopropylamine | pH 9.0, recombinant mutant R253K | Escherichia coli | |
0.15 | - |
8-Amino-7-oxononanoate | pH 9.0, recombinant mutant Y144F | Escherichia coli | |
0.3 | - |
S-adenosyl-L-methionine | pH 9.0, recombinant wild-type enzyme | Escherichia coli | |
0.75 | - |
S-adenosyl-(5')-3-methylthiopropylamine | pH 9.0, recombinant wild-type enzyme | Escherichia coli | |
1 | - |
S-adenosyl-L-methionine | pH 9.0, recombinant mutant R253A | Escherichia coli | |
3 | - |
S-adenosyl-L-methionine | pH 9.0, recombinant mutant Y144F | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
S-adenosyl-L-methionine + 8-amino-7-oxononanoate | Escherichia coli | second of 4 steps in the biosynthesis of vitamin H, i.e. biotin | S-adenosyl-4-methylthio-2-oxobutanoate + (7S,8R)-diaminononanoate | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P12995 | - |
- |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
S-adenosyl-L-methionine + 8-amino-7-oxononanoate = S-adenosyl-4-(methylsulfanyl)-2-oxobutanoate + 7,8-diaminononanoate | reaction mechanism, substrate binding structure, detailed reversible aminotransferase half-reaction, overview, residues Tyr17, Asp147, and conserved Tyr144 and Arg253 are important for catalysis | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
S-adenosyl-(5')-3-methylthiopropylamine + 8-amino-7-oxononanoate | decarboxylated S-adenosyl-L-methionine is as reactive as S-adenosyl-L-methionine | Escherichia coli | ? + (7S,8R)-diaminononanoate | - |
r | |
S-adenosyl-L-methionine + 8-amino-7-oxononanoate | second of 4 steps in the biosynthesis of vitamin H, i.e. biotin | Escherichia coli | S-adenosyl-4-methylthio-2-oxobutanoate + (7S,8R)-diaminononanoate | - |
r | |
S-adenosyl-L-methionine + 8-amino-7-oxononanoate | the aminotransferase half-reaction is reversible, substrate binding structure | Escherichia coli | S-adenosyl-4-methylthio-2-oxobutanoate + (7S,8R)-diaminononanoate | - |
r |
Synonyms | Comment | Organism |
---|---|---|
7,8-diaminopelargonic acid synthase | - |
Escherichia coli |
DAPA synthase | - |
Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | single turnover reactions | Escherichia coli | |
0.001 | - |
S-adenosyl-L-methionine | pH 9.0, recombinant mutant R253A | Escherichia coli | |
0.0035 | - |
S-adenosyl-L-methionine | pH 9.0, recombinant mutant Y17F | Escherichia coli | |
0.009 | - |
S-adenosyl-L-methionine | pH 9.0, recombinant mutant R253K | Escherichia coli | |
0.01 | - |
S-adenosyl-L-methionine | pH 9.0, recombinant mutant R253M | Escherichia coli | |
0.012 | - |
8-Amino-7-oxononanoate | pH 9.0, recombinant mutant Y17F | Escherichia coli | |
0.016 | - |
S-adenosyl-L-methionine | pH 9.0, recombinant wild-type enzyme | Escherichia coli | |
0.016 | - |
S-adenosyl-(5')-3-methylthiopropylamine | pH 9.0, recombinant wild-type enzyme and mutant R253K | Escherichia coli | |
0.04 | - |
8-Amino-7-oxononanoate | pH 9.0, recombinant mutant Y144F | Escherichia coli | |
0.06 | - |
S-adenosyl-L-methionine | pH 9.0, recombinant mutant Y144F | Escherichia coli | |
0.074 | - |
S-adenosyl-L-methionine | pH 9.0, recombinant mutant R253Q | Escherichia coli | |
0.67 | - |
8-Amino-7-oxononanoate | pH 9.0, recombinant mutant R253M | Escherichia coli | |
0.79 | - |
8-Amino-7-oxononanoate | pH 9.0, recombinant wild-type enzyme | Escherichia coli | |
1.3 | - |
8-Amino-7-oxononanoate | pH 9.0, recombinant mutant R253A | Escherichia coli | |
1.3 | - |
8-Amino-7-oxononanoate | pH 9.0, recombinant mutant R253K | Escherichia coli | |
1.7 | - |
8-Amino-7-oxononanoate | pH 9.0, recombinant mutant R253Q | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
9 | - |
assay at | Escherichia coli |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
pyridoxal 5'-phosphate | i.e. vitamin B6, dependent on | Escherichia coli |