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Literature summary for 1.2.1.9 extracted from

  • Dong, J.; Kan, B.; Liu, H.; Zhan, M.; Wang, S.; Xu, G.; Han, R.; Ni, Y.
    CRISPR-Cpf1-assisted engineering of Corynebacterium glutamicum SNK118 for enhanced L-ornithine production by NADP-dependent glyceraldehyde-3-phosphate dehydrogenase and NADH-dependent glutamate dehydrogenase (2020), Appl. Biochem. Biotechnol., 191, 955-967 .
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
gene gapC, recombinant expression in Corynebacterium glutamicum strain KBJ11, method optimization, coexpression with gene BsrocG encoding NADH-dependent glutamate dehydrogenase from Bacillus subtilis strain HB-1, recombinant expression in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain JM109 Clostridium saccharobutylicum
gene gapC, recombinant expression in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain JM109 Clostridium beijerinckii
gene gapC, recombinant expression in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain JM109 Clostridium acetobutylicum

Protein Variants

Protein Variants Comment Organism
additional information CRISPR-Cpf1-assisted engineering of Corynebacterium glutamicum SNK118 for enhanced L-ornithine production by NADP-dependent glyceraldehyde-3-phosphate dehydrogenase and NADH-dependent glutamate dehydrogenase. Insufficient intracellular NADPH content has been recognized as a limiting factor in L-ornithine biosynthesis. Overexpression of NADP-dependent glyceraldehyde 3-phosphate dehydrogenase gene CsgapC from Clostridium saccharobutylicum strain DSM 13864 and NADH-dependent glutamate dehydrogenase gene BsrocG from Bacillus subtilis strain HB-1 results in markedly increased ornithine biosynthesis. The engineered Corynebacterium glutamicum strain KBJ11 (SNK118DELTAargRDELTAargFDELTAncgl2228/pXMJ19-CsgapC-BsrocG) is constructed for L-ornithine overproduction. In fed-batch fermentation, L-ornithine of 88.26 g/l with productivity of 1.23 g/l/h (over 72 h) and yield of 0.414 g/g glucose is achieved by strain KBJ11 in a 10-l bioreactor. High titer and yield of L-ornithine production by microbial fermentation proves that heterologous expression of CsgapC and BsrocG can promote L-ornithine production by Corynebacterium glutamicum strains Clostridium saccharobutylicum

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
D-glyceraldehyde 3-phosphate + NADP+ + H2O Clostridium acetobutylicum
-
3-phospho-D-glycerate + NADPH + 2 H+
-
?
D-glyceraldehyde 3-phosphate + NADP+ + H2O Clostridium saccharobutylicum
-
3-phospho-D-glycerate + NADPH + 2 H+
-
?
D-glyceraldehyde 3-phosphate + NADP+ + H2O Clostridium acetobutylicum 1.7
-
3-phospho-D-glycerate + NADPH + 2 H+
-
?
D-glyceraldehyde 3-phosphate + NADP+ + H2O Clostridium saccharobutylicum DSM 13864
-
3-phospho-D-glycerate + NADPH + 2 H+
-
?

Organism

Organism UniProt Comment Textmining
Clostridium acetobutylicum
-
-
-
Clostridium acetobutylicum 1.7
-
-
-
Clostridium beijerinckii
-
-
-
Clostridium beijerinckii DSM 1739
-
-
-
Clostridium saccharobutylicum
-
-
-
Clostridium saccharobutylicum DSM 13864
-
-
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [┬Ámol/min/mg] Specific Activity Maximum [┬Ámol/min/mg] Comment Organism
2
-
purified recombinant enzyme, pH 8.5, 30┬░C Clostridium acetobutylicum
5.263
-
purified recombinant enzyme, pH 8.5, 30┬░C Clostridium saccharobutylicum

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-glyceraldehyde 3-phosphate + NADP+ + H2O
-
Clostridium acetobutylicum 3-phospho-D-glycerate + NADPH + 2 H+
-
?
D-glyceraldehyde 3-phosphate + NADP+ + H2O
-
Clostridium saccharobutylicum 3-phospho-D-glycerate + NADPH + 2 H+
-
?
D-glyceraldehyde 3-phosphate + NADP+ + H2O
-
Clostridium acetobutylicum 1.7 3-phospho-D-glycerate + NADPH + 2 H+
-
?
D-glyceraldehyde 3-phosphate + NADP+ + H2O
-
Clostridium saccharobutylicum DSM 13864 3-phospho-D-glycerate + NADPH + 2 H+
-
?
additional information recombinant CbgapC shows no catalytic activity Clostridium beijerinckii ?
-
-
additional information recombinant CbgapC shows no catalytic activity Clostridium beijerinckii DSM 1739 ?
-
-

Synonyms

Synonyms Comment Organism
CagapC
-
Clostridium acetobutylicum
CbgapC
-
Clostridium beijerinckii
CsgapC
-
Clostridium saccharobutylicum
GAPC
-
Clostridium beijerinckii
GAPC
-
Clostridium acetobutylicum
GAPC
-
Clostridium saccharobutylicum
NADP-dependent glyceraldehyde 3-phosphate dehydrogenase
-
Clostridium beijerinckii
NADP-dependent glyceraldehyde 3-phosphate dehydrogenase
-
Clostridium acetobutylicum
NADP-dependent glyceraldehyde 3-phosphate dehydrogenase
-
Clostridium saccharobutylicum

Temperature Optimum [┬░C]

Temperature Optimum [┬░C] Temperature Optimum Maximum [┬░C] Comment Organism
30
-
assay at Clostridium beijerinckii
30
-
assay at Clostridium acetobutylicum
30
-
assay at Clostridium saccharobutylicum

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.5
-
assay at Clostridium beijerinckii
8.5
-
assay at Clostridium acetobutylicum
8.5
-
assay at Clostridium saccharobutylicum

Cofactor

Cofactor Comment Organism Structure
NADP+
-
Clostridium beijerinckii
NADP+
-
Clostridium acetobutylicum
NADP+
-
Clostridium saccharobutylicum