Application | Comment | Organism |
---|---|---|
synthesis | 2-methylsuccinic acid is a C5 branched-chain dicarboxylate that serves as an attractive synthon for the synthesis of polymers with extensive applications in coatings, cosmetic solvents and bioplastics. A promising platform for 2-methylsuccinic acid bioproduction is established. Over-expression of codon-optimized citramalate synthase variant CimA from Methanococcus jannaschii, endogenous isopropylmalate isomerase EcLeuCD and enoate reductase YqjM from Bacillus subtilis allows the production of 2-methylsuccinic acid in Escherichia coli. Incorporation of the enzyme (KpnER) into the 2-methylsuccinic acid biosynthetic pathway leads to 2-methylsuccinic acid production improvement to a titer of 0.96 g/l in aerobic condition | Klebsiella pneumoniae |
synthesis | 2-methylsuccinic acid is a C5 branched-chain dicarboxylate that serves as an attractive synthon for the synthesis of polymers with extensive applications in coatings, cosmetic solvents and bioplastics. A promising platform for 2-methylsuccinic acid bioproduction is established. Over-expression of codon-optimized citramalate synthase variant CimA from Methanococcus jannaschii, endogenous isopropylmalate isomerase EcLeuCD and enoate reductase YqjM from Bacillus subtilis allows the production of 2-methylsuccinic acid in Escherichia coli. Incorporation of the enzyme (KpnER) into the 2-methylsuccinic acid biosynthetic pathway leads to 2-methylsuccinic acid production improvement to a titer of 0.96 g/l in aerobic condition | Bacillus subtilis |
Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli with N-terminal His6-tag | Klebsiella pneumoniae |
expression in Escherichia coli with N-terminal His6-tag | Bacillus subtilis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00259 | - |
NADPH | pH 7.5, 25°C | Klebsiella pneumoniae | |
0.01157 | - |
NADH | pH 7.5, 25°C | Klebsiella pneumoniae | |
0.039 | - |
NADPH | pH 7.5, 25°C | Bacillus subtilis | |
0.0587 | - |
NADH | pH 7.5, 25°C | Bacillus subtilis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus subtilis | P54550 | - |
- |
Bacillus subtilis 168 | P54550 | - |
- |
Klebsiella pneumoniae | A6T9B7 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Klebsiella pneumoniae |
- |
Bacillus subtilis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
citraconate + NADH + H+ | exhibits higher activity towards citraconate than NAD(P)H-dependent enoate reductase YqjM from Bacillus subtilis | Klebsiella pneumoniae | 2-methylsuccinic acid + NAD+ | - |
? | |
citraconate + NADH + H+ | exhibits lower activity towards citraconate than NAD(P)H-dependent enoate reductase KpnER from Klebsiella pneumoniae A6T9B7 | Bacillus subtilis | 2-methylsuccinic acid + NAD+ | - |
? | |
citraconate + NADH + H+ | exhibits lower activity towards citraconate than NAD(P)H-dependent enoate reductase KpnER from Klebsiella pneumoniae A6T9B7 | Bacillus subtilis 168 | 2-methylsuccinic acid + NAD+ | - |
? | |
citraconate + NADPH + H+ | exhibits higher activity towards citraconate than NAD(P)H-dependent enoate reductase YqjM from Bacillus subtilis | Klebsiella pneumoniae | 2-methylsuccinic acid + NADP+ | - |
? | |
citraconate + NADPH + H+ | exhibits lower activity towards citraconate than NAD(P)H-dependent enoate reductase KpnER from Klebsiella pneumoniae A6T9B7 | Bacillus subtilis | 2-methylsuccinic acid + NADP+ | - |
? | |
citraconate + NADPH + H+ | exhibits lower activity towards citraconate than NAD(P)H-dependent enoate reductase KpnER from Klebsiella pneumoniae A6T9B7 | Bacillus subtilis 168 | 2-methylsuccinic acid + NADP+ | - |
? |
Synonyms | Comment | Organism |
---|---|---|
KpnER | - |
Klebsiella pneumoniae |
NAD(P)H-dependent enoate reductase | - |
Klebsiella pneumoniae |
NAD(P)H-dependent enoate reductase | - |
Bacillus subtilis |
YqjM | - |
Bacillus subtilis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0153 | - |
NADH | pH 7.5, 25°C | Bacillus subtilis | |
0.0168 | - |
NADPH | pH 7.5, 25°C | Bacillus subtilis | |
0.274 | - |
NADH | pH 7.5, 25°C | Klebsiella pneumoniae | |
0.278 | - |
NADPH | pH 7.5, 25°C | Klebsiella pneumoniae |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADH | the enzyme utilizes both NADH and NADPH as reductants | Klebsiella pneumoniae | |
NADH | the enzyme utilizes both NADH and NADPH as reductants | Bacillus subtilis | |
NADPH | the enzyme utilizes both NADH and NADPH as reductants | Klebsiella pneumoniae | |
NADPH | the enzyme utilizes both NADH and NADPH as reductants | Bacillus subtilis |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.26 | - |
NADH | pH 7.5, 25°C | Bacillus subtilis | |
0.43 | - |
NADPH | pH 7.5, 25°C | Bacillus subtilis | |
23.68 | - |
NADH | pH 7.5, 25°C | Klebsiella pneumoniae | |
107.3 | - |
NADPH | pH 7.5, 25°C | Klebsiella pneumoniae |