产几丁质酶菌株Acinetobacter sp. CZW011的筛选鉴定与酶学性质研究  

Screening,identification and enzymatic properties of Acinetobacter sp. CZW011 strain produced by chitinase

作　　者：宋阳 陈梦 吴新财 丁志雯 李甜 刘耀东 房耀维 刘姝 SONG Yang;CHEN Meng;WU Xincai;DING Zhiwen;LI Tian;LIU Yaodong;FANG Yaowei;LIU Shu(College of Food Science and Engineering,Jiangsu Ocean University,Lianyungang 222005;Comprehensive Center of Lianyungang for Quality and Technical Inspection and Testing,Lianyungang 222006;Lianyungang Food and Drug Inspection and Testing Center,222000)

机构地区：[1]江苏海洋大学食品科学与工程学院,连云港222005 [2]连云港市质量技术综合检验检测中心,连云港222006 [3]连云港市食品药品检验检测中心,222000

出　　处：《中国食品添加剂》2022年第11期1-8,共8页China Food Additives

基　　金：国家自然科学基金项目(31772016);江苏省高等学校自然科学研究重大项目(20KJA550001);江苏省第五期“333工程”科研项目(BRA2019243);江苏省大学生创新项目(Z202011641631020、Z202011641633009);江苏省研究生科研与实践创新计划项目(KYCX20_2883)。

摘　　要：几丁质的自然资源十分丰富,因此新型产几丁质酶菌株的发现具有重要的研究价值。本研究是从连云港连岛黄海海域海泥中筛选获得的一株产几丁质酶菌株,并对所筛选的菌株进行菌株鉴定,完成其酶学性质实验。采用胶体几丁质平板产生透明圈法从海泥样品中筛选出产几丁质酶的细菌,获得产几丁质酶水平较高的菌株CZW011,并进一步通过摇瓶发酵,对菌株CZW011的酶学性质进行研究。培养菌株CZW011,光学显微镜下观察其形态学特征,完成生理生化试验,并将PCR产物完成序列扩增及分析,可鉴定CZW011菌株为不动杆菌属。通过研究菌株CZW011酶学性质发现,在温度35℃,pH=7.0的条件下水解几丁质能力最强。在25~35℃孵育2h,残留酶活高于80%;在pH 6.0~7.0孵育2h,残留酶活高于90%,是一种中温中性几丁质酶。常规金属离子Zn^(2+)、Ca^(2+)对酶活无明显促进或抑制的作用,Mg^(2+)有明显促进作用,Ag^(+)对酶活有明显的抑制作用。本菌株的发现与研究丰富了产几丁质酶菌株数据库,为几丁质酶的实际应用提供了一个新的思路。Chitin is rich in natural.New chitinase producing strains have important research value.A chitinase production strain was discovered from sea mud of Lianyungang Lian lsland,it was screened and identified in order to study its enzymatic properties.The method of producing a transparent circle on the colloidal chitin plate was applied to obtain the strain CZW011,and enzymology of the strain was further analyzed by shake flask fermentation.The strain CZW011 was cultured,its morphological characteristics were observed under a microscope,the physiological and biochemical tests were completed,and sequence amplification and analysis of PCR product was carried out.The CZW011 strain was identified as Acinetobacter sp.The optimum temperature of Acinetobacter sp.CZW011 to hydrolyze chitin was 35℃,and the optimum pH was pH 7.0.After being incubated at 25~35℃for 2h,the residual enzyme activity was higher than 80%;after being incubated at pH 7.0~8.0 for 2h,the residual enzyme activity was higher than 90%.Conventional metal ions Zn^(+),Ca^(2+) had no significant inhibitory effect on the enzyme activity,Mg^(2+) had a significant promoting effect,and Ag^(+) had significant inhibitory effect on the enzyme activity.The discovery and research of this strain enriches the database of chitinase-producing strains and provide a new idea for the application of chitinase.

关 键 词：几丁质酶 Acinetobacter sp. 鉴定 酶活 

分 类 号：TS202.1[轻工技术与工程—食品科学] TS201.3[轻工技术与工程—食品科学与工程]