Engineering of coenzyme specificity of formate dehydrogenase from Saccharomyces cerevisiae
- 1 November 2002
- journal article
- Published by Portland Press Ltd. in Biochemical Journal
- Vol. 367 (3) , 841-847
- https://doi.org/10.1042/bj20020379
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(+).Keywords
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