文献类型：期刊文献

英文题名：Computer-aided rational design strategy based on protein surface charge to improve the thermal stability of a novel esterase from Geobacillus jurassicus

作者：Song, Runfei[1];Zhang, Jin[1];Zhu, Mengyu[1];Lin, Lin[2,3];Wei, Wei[1];Wei, Dongzhi[1]

机构：[1]East China Univ Sci & Technol, Newworld Inst Biotechnol, State Key Lab Bioreactor Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China;[2]Shanghai Inst Technol, Sch Chem & Environm Engn, Shanghai 201418, Peoples R China;[3]Natl Engn Res Ctr Nanotechnol, Res Lab Funct Nanomat, Shanghai 200241, Peoples R China

年份：2024

卷号：46

期号：3

起止页码：443

外文期刊名：BIOTECHNOLOGY LETTERS

收录：;EI(收录号：20241315802499);WOS:【SCI-EXPANDED(收录号:WOS:001190299000001)】；

基金：This work was supported by the grant from the National Key Research and Development Program of China (2021YFC2100300), the Shanghai Natural Science Foundation (No. 20ZR1415400), the Shanghai Institute of Technology's collaborative innovation foundation (10120K228051), and the Shanghai Institute of Technology's scientific research start funds (10120K226107).

语种：英文

外文关键词：Surface charge protein engineering; Thermostable esterase; Site-directed mutagenesis; Rational design

摘要：Objectives Although Geobacillus are significant thermophilic bacteria source, there are no reports of thermostable esterase gene in Geobacillus jurassicus or rational design strategies to increase the thermal stability of esterases. Results Gene gju768 showed a highest similarity of 15.20% to esterases from Geobacillus sp. with detail enzymatic properties. Using a combination of Gibbs Unfolding Free Energy (Delta Delta G) calculator and the distance from the mutation site to the catalytic site (Ds(C alpha-C alpha)) to screen suitable mutation sites with elimination of negative surface charge, the mutants (D24N, E221Q, and E253Q) displayed stable mutants with higher thermal stability than the wild-type (WT). Mutant E253Q exhibited the best thermal stability, with a half-life (T-1/2) at 65(degrees)C of 32.4 min, which was 1.8-fold of the WT (17.9 min). Conclusion Cloning of gene gju768 and rational design based on surface charge engineering contributed to the identification of thermostable esterase from Geobacillus sp. and the exploration of evolutionary strategies for thermal stability.