高校化学工程学报2017，Vol.31Issue(3)：657-662,6.DOI:10.3969/j.issn.1003-9015.2017.03.021
活性位点邻近的Ω-loop对胰蛋白酶热稳定性和活性的影响
Effects of?-Loop Near Active Sites on the Stability and Activity of Trypsin
摘要
Abstract
Thermostability and activity of porcine trypsin were studied by deleting?-loop near active sites. Molecular dynamics software Gromacs v4.5.5 was used to predict Fragment 41~46 at the flexible region of porcine trypsin (which is the?-loop region). Subsequently, three mutations R1Q, K3Q and C5Q were obtained by deleting different amounts of amino acids in the?-loop. The results demonstrate that the activities of R1Q, K3Q and C5Q are all improved, and the maximum activity of R1Q is improved by 1.42 times compared to that of the wild type. Meanwhile, the inactivation half-life period and the optimal reaction temperature of R1Q are increased by 213.3 min and 11.8℃, respectively. Moreover, thermostability is improved for R1Q and K3Q, while it is decreased for C5Q. The results indicate that R1Q with isoleucine deleted from the?-loop shows both thermostability and activity improvements. This study provides a promising method to improve thermostability and activity of other industrial enzymes.关键词
猪源胰蛋白酶/Ω-loop/理性设计/热稳定性/活性Key words
porcine trypsin/Ω-loop/rational design method/thermostability/activity分类
生物科学引用本文复制引用
杜坤,甘一如,黄鹤..活性位点邻近的Ω-loop对胰蛋白酶热稳定性和活性的影响[J].高校化学工程学报,2017,31(3):657-662,6.基金项目
国家自然科学基金(31470967) （31470967）
国家科技重大专项(2011ZX09201-301-05,2014ZX09508006-002-002). （2011ZX09201-301-05,2014ZX09508006-002-002）
