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金枪鱼肉降解用高产蛋白酶菌株的筛选及鉴定
引用本文:吴丽娜,赵小惠,郭凯晴,杨宇杰,方旭波,陈小娥,张银照.金枪鱼肉降解用高产蛋白酶菌株的筛选及鉴定[J].湖北农业科学,2017,56(13).
作者姓名:吴丽娜  赵小惠  郭凯晴  杨宇杰  方旭波  陈小娥  张银照
作者单位:1. 浙江海洋大学食品与医药学院,浙江 舟山,316022;2. 浙江海洋大学食品与医药学院,浙江 舟山 316022;浙江省水产品加工技术研究联合重点实验室,浙江 舟山 316022;3. 浙江丰宇海洋生物制品有限公司,浙江 舟山,316104
基金项目:浙江省科技计划项目,国家级大学生创新创业训练计划项目
摘    要:采用透明圈法从舟山普陀白山土壤中筛选出3株能分泌胞外蛋白酶的菌株,并对蛋白酶活力最高的菌株进行分子鉴定及最适宜生长条件分析。选择福林酚试剂法测定其蛋白酶活力,对蛋白酶活力最高的菌株进行16S r RNA基因测序,用MEGA软件构建系统发育树,并考察其接种量、温度、时间对该菌株产酶活力的影响。结果表明,zjoub-1菌为高产蛋白酶菌属,并确定为枯草芽孢杆菌。该菌株用于金枪鱼暗色肉发酵时的最适宜条件为接种量2%、pH 7.4、温度37℃、时间48 h。在此生长条件下,菌株产蛋白酶活力最高,为49.21 U/m L,能够有效地降解金枪鱼暗色肉。

关 键 词:金枪鱼肉  透明圈法  福林酚试剂法  筛选  鉴定  生长条件

Screening and Identification of A Strain with High Protease in Fermentation of Tuna Dark Meat
WU Li-na,ZHAO Xiao-hui,GUO Kai-qing,YANG Yu-jie,FANG Xu-bo,CHEN Xiao-e,ZHANG Yin-zhao.Screening and Identification of A Strain with High Protease in Fermentation of Tuna Dark Meat[J].Hubei Agricultural Sciences,2017,56(13).
Authors:WU Li-na  ZHAO Xiao-hui  GUO Kai-qing  YANG Yu-jie  FANG Xu-bo  CHEN Xiao-e  ZHANG Yin-zhao
Abstract:Using Three strains of bacteria secreting extracellular protease were isolated from soil of Baishan in Zhoushan by transparent circle method, the bacterium with highest protease activity was identified and the most suitable growth conditions was studied. The collected samples were screened by transparent circle method to get three pure strains whose protease activ-ity would be measured by Forint phenol reagent method. The strain with the highest protease activity was identified by the 16S rRNA sequence analysis and the phylogenetic tree was constructed by MEGA software. The inoculation amount,tempera-ture,time and pH had been analyzed on the influence of its protease activity. The results showed that the study demonstrates that the zjoub-1 strain with highest protease activity was defined as bacillus subtilis. The optimal condition for fermentation of Tuna dark meat was as follows: the inoculation quantity was 2℅,the incubation temperature was 37℃, the incubation time was about 48 h, the initial pH was 7.4. Under this circumstance, the strain has the highest protease activity, which illus-trates that the strain with high protease activity can degrade the Tuna dark meat efficiently.
Keywords:Tuna dark meat  transparent circle method  Forint phenol reagent method  isolation  identification  growth conditions
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