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. 2002 Nov 1;367(Pt 3):841–847. doi: 10.1042/BJ20020379

Engineering of coenzyme specificity of formate dehydrogenase from Saccharomyces cerevisiae.

Alexander E Serov 1, Anna S Popova 1, Vladimir V Fedorchuk 1, Vladimir I Tishkov 1
PMCID: PMC1222933  PMID: 12144528

Abstract

A eukaryotic formate dehydrogenase (EC 1.2.1.2, FDH) with its substrate specificity changed from NAD(+) to NADP(+) has been constructed by introducing two single-point mutations, Asp(196)-->Ala (D196A) and Tyr(197)-->Arg (Y197R). The mutagenesis was based on the results of homology modelling of a NAD(+)-specific FDH from Saccharomyces cerevisiae (SceFDH) using the Pseudomonas sp.101 FDH (PseFDH) crystal structure as a template. The resulting model structure suggested that Asp(196) and Tyr(197) mediate the absolute coenzyme specificity of SceFDH for NAD(+).

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Selected References

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