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Literature summary extracted from

  • Wang, J.; Yang, Y.; Zhang, R.; Shen, X.; Chen, Z.; Wang, J.; Yuan, Q.; Yan, Y.
    Microbial production of branched-chain dicarboxylate 2-methylsuccinic acid via enoate reductase-mediated bioreduction (2018), Metab. Eng., 45, 1-10 .
    View publication on PubMed

Application

EC Number Application Comment Organism
1.3.1.31 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
1.3.1.31 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(Commentary)

EC Number Cloned (Comment) Organism
1.3.1.31 expression in Escherichia coli with N-terminal His6-tag Klebsiella pneumoniae
1.3.1.31 expression in Escherichia coli with N-terminal His6-tag Bacillus subtilis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.3.1.31 0.00259
-
NADPH pH 7.5, 25°C Klebsiella pneumoniae
1.3.1.31 0.01157
-
NADH pH 7.5, 25°C Klebsiella pneumoniae
1.3.1.31 0.039
-
NADPH pH 7.5, 25°C Bacillus subtilis
1.3.1.31 0.0587
-
NADH pH 7.5, 25°C Bacillus subtilis

Organism

EC Number Organism UniProt Comment Textmining
1.3.1.31 Bacillus subtilis P54550
-
-
1.3.1.31 Bacillus subtilis 168 P54550
-
-
1.3.1.31 Klebsiella pneumoniae A6T9B7
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.3.1.31
-
Klebsiella pneumoniae
1.3.1.31
-
Bacillus subtilis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.3.1.31 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+
-
?
1.3.1.31 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+
-
?
1.3.1.31 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+
-
?
1.3.1.31 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+
-
?
1.3.1.31 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+
-
?
1.3.1.31 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

EC Number Synonyms Comment Organism
1.3.1.31 KpnER
-
Klebsiella pneumoniae
1.3.1.31 NAD(P)H-dependent enoate reductase
-
Klebsiella pneumoniae
1.3.1.31 NAD(P)H-dependent enoate reductase
-
Bacillus subtilis
1.3.1.31 YqjM
-
Bacillus subtilis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.3.1.31 0.0153
-
NADH pH 7.5, 25°C Bacillus subtilis
1.3.1.31 0.0168
-
NADPH pH 7.5, 25°C Bacillus subtilis
1.3.1.31 0.274
-
NADH pH 7.5, 25°C Klebsiella pneumoniae
1.3.1.31 0.278
-
NADPH pH 7.5, 25°C Klebsiella pneumoniae

Cofactor

EC Number Cofactor Comment Organism Structure
1.3.1.31 NADH the enzyme utilizes both NADH and NADPH as reductants Klebsiella pneumoniae
1.3.1.31 NADH the enzyme utilizes both NADH and NADPH as reductants Bacillus subtilis
1.3.1.31 NADPH the enzyme utilizes both NADH and NADPH as reductants Klebsiella pneumoniae
1.3.1.31 NADPH the enzyme utilizes both NADH and NADPH as reductants Bacillus subtilis

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.3.1.31 0.26
-
NADH pH 7.5, 25°C Bacillus subtilis
1.3.1.31 0.43
-
NADPH pH 7.5, 25°C Bacillus subtilis
1.3.1.31 23.68
-
NADH pH 7.5, 25°C Klebsiella pneumoniae
1.3.1.31 107.3
-
NADPH pH 7.5, 25°C Klebsiella pneumoniae