... and Asp245, to coenzymebinding and specificity in P. asaccharolyticus GDH.Five mutants were created: in E243K and E243R, theP7 glutamate was replaced by positively charged lysine and arginine, ... coenzyme binding was tighter and thereforemeasurable and, in particular, gave measurable Kdvalues for binding of NADPH to both E243K and E243R, whereas the binding of NADH to thesemutants was ... E243R and E243K, reflecting thecombination of improved efficiency with NADPH and diminished efficiency with NADH. This is muchgreater than the shift in discrimination values for D245K, E243D and...