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Engineering of coenzyme specificity of formate dehydrogenase from Saccharomyces cerevisiae.

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(+).[1]

References

  1. Engineering of coenzyme specificity of formate dehydrogenase from Saccharomyces cerevisiae. Serov, A.E., Popova, A.S., Fedorchuk, V.V., Tishkov, V.I. Biochem. J. (2002) [Pubmed]
 
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