摘要
讨论并提出了有效增强重组腈代谢酶系表达活性的多种策略。根据所选用的载体和宿主的不同表达特性,同时克隆目的基因上、下游的正向调控序列可直接增强酶表达水平;强化翻译后的修饰,可有效提高活性;除了常用的大肠杆菌克隆表达体系外,开发新型宿主-载体系统可为重组酶的活性表达提供新的途径;随着生物信息学技术的迅速发展,利用已知三维结构的蛋白对目标酶进行突变位点设计及结构与功能预测,将有望加快腈代谢酶系的定点突变研究进程。
To enhance the functional expression of nitriledegradation enzymatic system,several effiecient strategies are presented and discussed.According to the different expression characteristics of the clone vector and the host,the expression level of the enzyme can be directly raised by cloning the upstream or downstream genetic sequences of the positive regulators simultaneously.The posttranslational modification being strengthened,the activity can be raised efficiently.The development of novel hostvector systems can provide a new approach for the functional expression of the recombination enzyme.Based on the knowledge of the 3D stucture of the enzyme,the mutational site design and the stucture and function prediction can be made with the rapid development of the bioinformatics,which in turn will accelerate the research progress in the sitedirected mutagenesis of the nitriledegradation enzymatic system.
出处
《现代化工》
CAS
CSCD
北大核心
2003年第9期23-26,28,共5页
Modern Chemical Industry
基金
国家自然科学基金资助项目(20206014)
关键词
腈水合酶
腈代谢酶系
表达活性
载体
重组酶
nitrile hydratase
nitrile-degradation enzymatic system
functional expression