松毛虫肠道产蛋白酶菌株的筛选鉴定及培养条件研究-中国农学通报.pdfVIP

中国农学通报 2012,28(14): 18-21 Chinese Agricultural Science Bulletin 松毛虫肠道产蛋白酶菌株的筛选鉴定及培养条件研究 孙佑赫，周开艳，熊 智 （西南林业大学林学院，昆明650224） 摘 要：为了获得产蛋白酶能力高的细菌资源，采用酪蛋白平板法从松毛虫肠道中分离筛选得到16株产 蛋白酶的菌株。通过比较透明圈大小和测定发酵后酶活力，筛选出一株产酶能力较高的菌株2N0 1。根 据序列比对结果，构建关于2N0 1 的系统发育树，将其鉴定为Enterobacter hormaechei 。研究培养条件对 该菌株生长的影响，确定2N0 1 产酶培养的最适温度是40℃，最适pH 8.0 ，培养72 h 左右出现产酶峰值， 酶活力达50.07 U/mL 。通过研究表明，菌株2N0 1是产蛋白酶的较好材料，具有一定的应用前景。 关键词：肠道细菌；蛋白酶；鉴定；酶学性质 中图分类号：Q933 文献标志码：A 论文编号：2011-3833 ScreeningandIdentificationofProtease-producingBacteriumfromIntestinalCanal ofDendrolimuandLiquidCultureStudies Sun Youhe, Zhou Kaiyan, Xiong Zhi (FacultyofForestry,SouthwestForestryUniversity, Kunming 650224) Abstract: In order to breed high-yield strains for protease, sixteen protease-producing strains were isolated from intestinal canal of Dendrolimu houi by casein plate. 2N0 1 was screened as a higher-yield strain for protease by comparing the size of transparent circle and enzyme activity. A phylogenetic tree was constructed with the published 16S rDNA sequences of relative bacteria species. In the phylogenetic tree, the strain 2N0 1 T was in the branch with the Enterobacter hormaechei CIP 103441 (AJ508302). The most suitably culture conditions for the growth of strain 2N0 1 was temperature 40℃ and pH 8.0. The highest protease activity of strain 2N0 1 was 50.07 U/mL when grown for 72 h. This study provided a good experimental material for the further industrial utilization about protease production. Keywords:intestinal bacteria; protease; identificati