Alkaline phosphatase (AP) is a member of an enzyme superfamily capable of catalyzing phosphate and sulfate hydrolysis reactions for a variety of substrates. Catalytic promiscuity of E. coli AP has been previously studied and suggests a link between active site flexibility and increased cross superfamily catalytic activity. Typically, mutations of site Asp101 in E. coli AP cause a decrease in catalytic efficiency due to the residue’s proximity and stabilization of the neighboring alkoxide nucleophile Ser102. Interestingly, mutation of Asp101 to histidine yields a decrease in binding affinity and a concomitant increase in catalytic efficiency (kcat/KM). Further investigation into the mechanistic and structural variations caused by this mutation will be important in deciphering requirements for the evolution of a more promiscuous and catalytically efficient AP enzyme.

Designing a fast enzyme 101: D101H mutation of E. coli alkaline phosphatase

2019 Spring National Meeting

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