Change in Substrate Preference of Streptomyces Aminopeptidase through Modification of the Environment around the Substrate Binding Site | Zendy

Jiro Arima | Zendy; Yoshiko Uesugi | Zendy; Masaki Iwabuchi | Zendy; Tadashi Hatanaka | Zendy

Open Access

Change in Substrate Preference of Streptomyces Aminopeptidase through Modification of the Environment around the Substrate Binding Site

Author(s) -

Jiro Arima,

Yoshiko Uesugi,

Masaki Iwabuchi,

Tadashi Hatanaka

Publication year - 2006

Publication title -

applied and environmental microbiology

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 1.552

H-Index - 324

eISSN - 1070-6291

pISSN - 0099-2240

DOI - 10.1128/aem.01460-06

Subject(s) - substrate (aquarium) , hydrolysis , aminopeptidase , enzyme , mutant , streptomyces , derivative (finance) , biochemistry , chemistry , substrate specificity , stereochemistry , biology , bacteria , genetics , amino acid , leucine , gene , ecology , financial economics , economics

We attempted to alter the substrate preference of aminopeptidase fromStreptomyces 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 higherk cat values for hydrolysis ofp -nitroanilide derivatives than those of the wild type, respectively).

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