产蛋白酶海洋细菌Bacillus sp.4sp5分离鉴定及降解南极磷虾原肌球蛋白的研究  

Isolation and characterization of the protease-producing marine bacterium Bacillus sp.4sp5 and its degradation of Antarctic krill tropomyosin

作　　者：白玥 郝青芳 周俊栏 包秀丽 郝悦 亢新鑫 王淑军 BAI Yue;HAO Qing-Fang;ZHOU Jun-Lan;BAO Xiu-Li;HAO Yue;KANG Xin-Xin;WANG Shu-Jun(Key Laboratory of Marine Biological Resources and Environment of Jiangsu Province,Jiangsu Ocean University,Lianyungang 222005,China;Key Laboratory of Marine Biotechnology of Jiangsu Province,Jiangsu Ocean University,Lianyungang 222005,China)

机构地区：[1]江苏海洋大学,江苏省海洋生物资源与环境重点实验室,连云港222005 [2]江苏海洋大学,江苏省海洋生物技术重点实验室,连云港222005

出　　处：《食品安全质量检测学报》2024年第4期98-105,共8页Journal of Food Safety and Quality

基　　金：国家重点研发计划项目(2022YFC2805101);连云港市“521工程”科研资助项目;耐盐海洋水产品加工专用蛋白酶制剂及活性肽制备关键技术研究项目(LYG06521202107);江苏省科研实践创新项目(SJCX-JOU2296)。

摘　　要：目的从海洋样品中筛选产蛋白酶海洋细菌及可以降解南极磷虾原肌球蛋白(tropomyosin,TM)的蛋白酶。方法采用透明圈法筛选产蛋白酶菌株,并根据聚丙烯酰胺凝胶电泳(polyacrylamide gel electrophoresis,PAGE)和酶联免疫吸附测定法(enzyme-linked immunosorbent assay,ELISA)确定降解效果最佳菌株,对其产的蛋白酶的酶学特性及降解原肌球蛋白条件进行研究。结果最佳为菌株4sp5,经16SrDNA鉴定为Bacillus sp.。蛋白酶的最适作用温度和pH分别为60℃和9。Na^(+)和K^(+)对蛋白酶活有促进作用,其对有机溶剂敏感,表面活性剂对其影响较小。3种抑制剂均可抑制酶的活性。当加酶量为24U/mL时,7.0h能够有效地降解南极磷虾原肌球蛋白,经ELISA试剂盒测定,其致敏性消除率达98.63%。结论菌株4sp5及其产生的蛋白酶在南极磷虾原肌球蛋白脱敏作用中具有应用前景。Objective To screen protease-producing marine bacteria produced protaese which could degrade tropomyosin(TM)of krill Antarctic from marine samples Methods The protease-producing strains were screened by the transparent circle method,and the ability of hydrolyzed TM was determined by polyacrylamide gel electrophoresis(PAGE)and enzyme-linked immunosorbent assay(ELISA).The enzymatic characteristics of the protease and the degradation conditions of tropomyosin were studied.Results The best strain 4sp5 was identified as Bacillus sp.by 16S rDNA.The optimal temperature and pH of the protease were 60℃ and 9,respectively.Na and Kt promoted the activity of the protease,which was sensitive to organic solvents,and had little effect on surfactants.The 3 kinds of inhibitors inhibited the activity of the protease.When the enzyme dosage was 24 U/mL,it could effectively degrade Antarctic krill myosin for 7.0 h.The sensitivity elimination rate of the enzyme was determined by ELISA kit to reach 98.63%.Conclusion The protease produced by strain 4sp5 has application prospects in the desensitization of tropomyosin in krill.

关 键 词：原肌球蛋白 南极磷虾 海洋细菌 蛋白酶 脱敏作用 

分 类 号：TS254.1[轻工技术与工程—水产品加工及贮藏工程]