文献类型：期刊文献

英文题名：Identification and magnetic immobilization of a pyrophilous aspartic protease from Antarctic psychrophilic fungus

作者：Gao, Bei[1];He, Lei[1];Wei, Dongzhi[1];Zhang, Lujia[2,3]

机构：[1]East China Univ Sci & Technol, New World Inst Biotechnol, State Key Lab Bioreactor Engn, Shanghai, Peoples R China;[2]East China Normal Univ, Sch Mol Engn, Dept Chem, Shanghai Engn Res Ctr Mol Therapeut & New Drug De, 500 Dongchuan Rd, Shanghai 200062, Peoples R China;[3]NYU Shanghai, NYU ECNU Ctr Computat Chem, Shanghai, Peoples R China

年份：2018

卷号：42

期号：6

外文期刊名：JOURNAL OF FOOD BIOCHEMISTRY

收录：;EI(收录号：20184105928579);WOS:【SCI-EXPANDED(收录号:WOS:000456942100053)】；

基金：This work was funded by the National Key Research Program of China (No. 2016YFA0501701), National Natural Foundation of China (No. 31772007, and No. 31571786), and Natural Science Foundation of Shanghai (No. 16ZR1449500), Innovation Program of Shanghai Municipal Education Commission; Grant number: 201701070005E00020. We are grateful to Institute of Ocean and Earth Sciences, University of Malaya for offering the strain, Geomyces pannorum.

语种：英文

外文关键词：aspartic protease; bond specificity; cheese-making; magnetic immobilization

摘要：Aspartic protease is a versatile protease used in the food processing industry. A novel aspartic protease gene (P10) was cloned from an Antarctic psychrophilic fungus Geomyces pannorum and successfully expressed in Aspergillus oryzae when cultured at 20 degrees C. However, purified P10 exhibited optimal activity at 60 degrees C and retained approximately 80% activity at 50-70 degrees C. The Km and Vmax values for this protease toward BSA were 1.01 mg/ml and 4.4 x 10-2 mg/(ml min), respectively, with a specific activity of 585 U/mg. P10 showed broad substrate specificity, with an increased affinity for hydrolyzing kappa-casein than for alpha-casein and beta-casein, indicating a potential value for cheese-making. P10 also presented wide peptide bond specificity toward the oxidized insulin B chain with high affinity for the C-terminus. Furthermore, P10 was immobilized on iron oxide nanoparticles, wherein it displayed improved thermo-stability and pH tolerance. These results provide novel insights into psychrophilic fungal enzymes, suggesting P10 as a potential biocatalyst.