吉林农业大学学报2011，Vol.33Issue(4)：395-398,417,5.DOI:CNKI:22-1100/S.20110511.1646.001
微小毛霉凝乳酶的分泌表达、产物纯化及鉴定
Secretory Expression, Purification and Identification of Rennet from Mucor pusillus
摘要
Abstract
The rennet from Mucor pusillus is one of the major sources of tnicrobial rennet, but it has some defects compared with the bovine chymosin. Aspartic proteinases from fungi are usually more heat resistant . When these enzymes are used for cheese making, the residue of enzymatic activity after heat treatment (55 ℃) following milk coagulation may still exist and cause proteolysis during cheese storage, bitter taste, softness and lower yield of the cheese products. In order to develop a rennet product with suitable milk clotting properties, the gene of rennet was cloned from Mucor pusillus, and the expression vector pPIC9K/M was constructed. The plasmids of pPIC9K/M was linearized with Sac I , and transformed into Pichia pastoris GS115 competent cells. The rennet was secretory expressed in Pichia pastoris successfully , and a strong band at about 46 kD was shown by SDS - PAGE. Activity tests showed that the rennet activity in the culture supernatant was 311.8 U/mL. The purity of recombinant rennet reached 92% with a 51.89% activity recovery.关键词
凝乳酶；分泌表达；毕赤酵母Key words
rennet/ secretory expression/ Pichia pastoris分类
轻工纺织引用本文复制引用
王景会,李玉秋,郑丽,李倬琳,高岩,杨贞耐..微小毛霉凝乳酶的分泌表达、产物纯化及鉴定[J].吉林农业大学学报,2011,33(4):395-398,417,5.基金项目
国家“863”计划项目(2006AA10Z306)，现代农业产业技术体系建设专项[农科教发(200714)号] （2006AA10Z306）
