Literature summary for 1.1.1.44 extracted from

Chen, H.; Zhu, Z.; Huang, R.; Zhang, Y.P.
Coenzyme engineering of a hyperthermophilic 6-phosphogluconate dehydrogenase from NADP(+) to NAD(+) with its application to biobatteries (2016), Sci. Rep., 6, 36311 .

Search for more literature for 1.1.1.44

Application

Application	Comment	Organism
additional information	the enzyme can be used for power production in biobatteries. Mutant N32E/R33I/T34I versus the wild-type 6PGDH are evaluated electrochemically in an anodic reaction system containing two enzymes: 6PGDH and diaphorase, a coenzyme (NADP+ or NAD+), an electron mediator AQDS, and a 6-phosphogluconate substrate. Cyclic voltammetry results clearly show that both enzymes produce significant oxidation current peaks at -0.3 V versus Ag/AgCl. The mutant N32E/R33I/T34I exhibits a current density 25% higher than that generated by the wild-type	Thermotoga maritima

Cloned(Commentary)

Cloned (Comment)	Organism
gene 6pgdh, recombinant overexpression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)	Thermotoga maritima

Protein Variants

Protein Variants	Comment	Organism
additional information	construction and evaluation of enzyme mutants with increased activity for NAD* compared to the wild-type enzyme	Thermotoga maritima
N32D	site-directed mutagenesis, mutant N32D has a 460times higher Km value for NADP+ and a slightly decreased Km value for NAD+, but no significant difference of kcat for NADP+, compared to the wild-type enzyme	Thermotoga maritima
N32D/R33I/T34I	site-directed mutagenesis, the triple mutant exhibits a far higher Km value for NADP+ and a slightly decreased Km value for NAD+, but no significant difference of kcat for NADP+, compared to the wild-type enzyme	Thermotoga maritima
N32D/R33L/T34S	site-directed mutagenesis, the triple mutant has a increased Km value for NADP+ and NAD+, but no significant difference of kcat for NADP+, compared to the wild-type enzyme	Thermotoga maritima
N32E/R33I/T34I	site-directed mutagenesis, the mutant shows almost 2fold declined Km and a 2fold increased kcat for NAD+ compared to wild-type, the catalytic efficiency kcat/Km towards NADP+ decreases, while the catalytic efficiency towards NAD+ increases	Thermotoga maritima

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
6-phospho-D-gluconate + NADP+	Thermotoga maritima	-	D-ribulose 5-phosphate + CO2 + NADPH + H+	-	?

Organism

Organism	UniProt	Comment	Textmining
Thermotoga maritima	Q9WYR9	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography to homogeneity	Thermotoga maritima

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
6-phospho-D-gluconate + NAD+	-	Thermotoga maritima	D-ribulose 5-phosphate + CO2 + NADH + H+	-	?
6-phospho-D-gluconate + NADP+	-	Thermotoga maritima	D-ribulose 5-phosphate + CO2 + NADPH + H+	-	?

Cofactor

Cofactor	Comment	Organism	Structure
NAD+	active with enzyme mutants, not with the wild-type enzyme	Thermotoga maritima
NADP+	specific for, natural coenzyme is NADP+	Thermotoga maritima

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Release 2023.1 (January 2023)