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Literature summary extracted from

  • Wang, Y.; San, K.Y.; Bennett, G.N.
    Improvement of NADPH bioavailability in Escherichia coli by replacing NAD+-dependent glyceraldehyde-3-phosphate dehydrogenase GapA with NADP+-dependent GapB from Bacillus subtilis and addition of NAD kinase (2013), J. Ind. Microbiol. Biotechnol., 40, 1449-1460 .
    View publication on PubMed

Application

EC Number Application Comment Organism
1.3.1.103 analysis monitoring the production of (S)-2-chloropropionate from 2-chloroacrylate as a model system for monitoring NADPH availability. A phosphofructokinase pfkA pfkB double-deletion strain shows the highest yield of 2-chloropropionic acid product. The flux distribution of fructose-6-phosphate between glycolysis and the pentose phosphate pathway determines the amount of NAPDH available for reductive biosynthesis Burkholderia sp.

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.2.1.13 gene gapB, recombinant overexpression in and complementation of Escherichia coli gapA-deficient strain MG1655 gapA::Tn10, gene gapB is expressed from plasmid pDHC29 under the Plac promoter control, coexpression with Escherichia coli gene nadK encoding the NAD kinase, genes gapB and nadk are grouped as a single operon Bacillus subtilis
1.3.1.103 expression in Escherichia coli Burkholderia sp.

Protein Variants

EC Number Protein Variants Comment Organism
1.2.1.13 additional information to increase NADPH bioavailability, the native NAD+-dependent glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gapA gene, EC 1.2.1.12, in Escherichia coli is replaced with the NADP+-dependent gapB gene, EC 1.2.1.13, from Bacillus subtilis. To overcome the limitation of NADP+ availability, Escherichia coli NAD kinase, gene nadK is also coexpressed with gapB in Escherichia coli. replacing NAD+-dependent GapA activity with NADP+-dependent GapB activity increases the synthesis of NADPH-dependent compounds Bacillus subtilis

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.2.1.13 D-glyceraldehyde 3-phosphate + phosphate + NADP+ Bacillus subtilis
-
 3-phospho-D-glyceroyl phosphate + NADPH + H+
-
 ?
1.2.1.13 D-glyceraldehyde 3-phosphate + phosphate + NADP+ Bacillus subtilis 168
-
 3-phospho-D-glyceroyl phosphate + NADPH + H+
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.2.1.13 Bacillus subtilis O34425
-
-
1.2.1.13 Bacillus subtilis 168 O34425
-
-
1.3.1.103 Burkholderia sp. Q59I44
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.2.1.13 D-glyceraldehyde 3-phosphate + phosphate + NADP+
-
 Bacillus subtilis 3-phospho-D-glyceroyl phosphate + NADPH + H+
-
 ?
1.2.1.13 D-glyceraldehyde 3-phosphate + phosphate + NADP+
-
 Bacillus subtilis 168 3-phospho-D-glyceroyl phosphate + NADPH + H+
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.2.1.13 GapB
-
 Bacillus subtilis
1.2.1.13 NADP+-dependent gapB
-
 Bacillus subtilis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.2.1.13 37
-
 assay at Bacillus subtilis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.2.1.13 8
-
 assay at Bacillus subtilis

Cofactor

EC Number Cofactor Comment Organism Structure
1.2.1.13 NADP+
-
 Bacillus subtilis