2020
DOI: 10.1101/2020.04.02.022350
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In vivoselection for formate dehydrogenases with high efficiency and specificity towards NADP+

Abstract: CGAR) 2 A b s t r a c t Efficient regeneration of cofactors is vital for the establishment of continuous biocatalytic processes.Formate is an ideal electron donor for cofactor regeneration due to its general availability, low reduction potential, and benign byproduct (CO 2 ). However, formate dehydrogenases (FDHs) are usual specific to NAD + , such that NADPH regeneration with formate is challenging. Previous studies reported naturally occurring FDHs or engineered FDHs that accept NADP + , but these enzymes sh… Show more

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Cited by 2 publications
(2 citation statements)
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“…26 One unit (U) of enzyme activity was defined as the amount of enzyme catalyzing the conversion of 1 μmol of substrate per minute under standard conditions. According to previous reports, 27,28 the kinetic constants were measured by varying the ammonium formate from 5 to 250 mM, with the NADP + (5−10) were used to determine the optimum pH for ApFDH and its variants. Citric acid−sodium citrate buffer (100 mM) was prepared for pH 5.0−6.0, phosphate buffer (100 mM) was prepared for pH 6.0−8.0, Tris-HCl buffer (100 mM) was prepared for pH 8.0−9.0, and glycine−NaOH buffer (100 mM) was prepared for pH 9.0−10.0.…”
Section: ■ Materials and Methodsmentioning
confidence: 99%
“…26 One unit (U) of enzyme activity was defined as the amount of enzyme catalyzing the conversion of 1 μmol of substrate per minute under standard conditions. According to previous reports, 27,28 the kinetic constants were measured by varying the ammonium formate from 5 to 250 mM, with the NADP + (5−10) were used to determine the optimum pH for ApFDH and its variants. Citric acid−sodium citrate buffer (100 mM) was prepared for pH 5.0−6.0, phosphate buffer (100 mM) was prepared for pH 6.0−8.0, Tris-HCl buffer (100 mM) was prepared for pH 8.0−9.0, and glycine−NaOH buffer (100 mM) was prepared for pH 9.0−10.0.…”
Section: ■ Materials and Methodsmentioning
confidence: 99%
“…Several studies also constructed mutant libraries of specific oxidoreductases and harnessed natural selection to identify variants with altered cofactor specificity [5][6][7]. Yet, until now, no study has attempted to systematically explore the overall evolvability of central metabolism oxidoreductases towards the use of a different cofactor.…”
Section: Introductionmentioning
confidence: 99%