通过修饰底物结合位点周围的环境改变链霉菌氨肽酶的底物偏好性。
Change in substrate preference of Streptomyces aminopeptidase through modification of the environment around the substrate binding site.
作者信息
Arima Jiro, Uesugi Yoshiko, Iwabuchi Masaki, Hatanaka Tadashi
机构信息
Research Institute for Biological Sciences (RIBS), Okayama, 7549-1 Kibichuo-cho, Kaga-gun, Okayama 716-1241, Japan.
出版信息
Appl Environ Microbiol. 2006 Dec;72(12):7962-7. doi: 10.1128/AEM.01460-06. Epub 2006 Oct 6.
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).
摘要
我们试图改变来自龟裂链霉菌TH-2（SSAP）的氨肽酶的底物偏好性。由于SSAP的Asp198和Phe221位于底物结合位点，我们筛选了2000种具有D198X/F221X突变的突变酶。通过进行这项检测，我们获得了两种酶；一种特异性水解精氨酰衍生物，另一种特异性水解胱氨酰衍生物（对硝基苯胺衍生物水解的k(cat)值分别比野生型高65倍和12.5倍）。
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