Engineering of coenzyme specificity of formate dehydrogenase from Saccharomyces cerevisiae
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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, Asp196→Ala (D196A) and Tyr197→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 Asp196 and Tyr197 mediate the absolute coenzyme specificity of SceFDH for NAD+.
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2012 ◽
Vol 25
(11)
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pp. 781-788
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2014 ◽
Vol 447
(4)
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pp. 626-632
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2004 ◽
Vol 64
(1)
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pp. 1-9
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2006 ◽
Vol 340
(3)
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pp. 792-799
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