Oct 10, 2006

Change in substrate preference of Streptomyces aminopeptidase through modification of the environment around the substrate binding site

Applied and Environmental Microbiology
Jiro ArimaTadashi Hatanaka

Abstract

We attempted to alter the substrate preference of aminopeptidase from Streptomyces septatus TH-2 (SSAP). Because Asp198 and Phe221 of SSAP are located in the substrate binding site, we screened 2,000 mutant enzymes with D198X/F221X mutations. By carrying out this examination, we obtained two enzymes; one specifically hydrolyzed an arginyl derivative, and the other specifically hydrolyzed a cystinyl derivative (65- and 12.5-fold higher k(cat) values for hydrolysis of p-nitroanilide derivatives than those of the wild type, respectively).

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Derivatives
Enzymes, antithrombotic
Streptomyces septatus
Aminopeptidase
Ligand Binding Domain
Substrate Specificity
Enzymes for Treatment of Wounds and Ulcers
Biotechnology
Mutant
Enzymes, hematological

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