Construction and Enzyme-activity Assessment of L-Asparaginase Mutants
- VernacularTitle:L-门冬酰胺酶突变体的构建及其活性测定
- Author:
Ruibo JIA
;
Jianhua CHEN
;
Yujun WEI
;
Xiangdong GAO
;
Wutong WU
- Publication Type:Journal Article
- Keywords:
L-Asparaginase;
Site-directed mutagenesis;
Overlap extension PCR;
Alanine-scanning
- From:
Journal of China Pharmaceutical University
2005;(5):468-472
- CountryChina
- Language:Chinese
-
Abstract:
AIM:To construct nine novel L-asparaginase mutants and study their enzyme-activity.METHODS:The mutants were constructed using overlap extension PCR according to the principle of alanine-scanning mutagenesis. The enzyme-activity was detected by Nessler's method. RESULTS:The DNA sequencing showed that the mutagenesis was consistent with the theoretical prediction. The enzyme-activity assay demonstrated that each mutant possessed enzyme activity equal to the original enzyme. CONCLUSION:Through gene modification,epitop of L-asparaginase was changed without activity loss.These results provide foundation for further study of the structure-function relationship of L-asparaginase.
