Identification of a chitinase from the hepatopancreas of Chinese black sleeper(Bostrychus sinensis)

作     者：Yulei Chen Zhipeng Tao Minghui Zhang Lechang Sun Guangming Liu Minjie Cao Yulei Chen;Zhipeng Tao;Minghui Zhang;Lechang Sun;Guangming Liu;Minjie Cao

作者机构：College of Food and Biological EngineeringJimei UniversityXiamen 361021China Fujian Collaborative Innovation Center for Exploitation and Utilization of Marine Biological ResourcesXiamen 361102China 

出 版 物：《Acta Oceanologica Sinica》 (海洋学报（英文版）)

年 卷 期：2021年第40卷第6期

页      面：50-60页

核心收录：

学科分类：0710[理学-生物学] 0908[农学-水产] 0707[理学-海洋科学] 09[农学] 

基　　金：The National Key R&D Program of China under contract No.2018YFD0901004 the National Natural Science Foundation of China under contract Nos 31772049,31702372 

主　　题：Bostrychus sinensis chitinase purification characterization molecular cloning chitosan 

摘      要：Chinese black sleeper(Bostrychus sinensis)is a fish that lives both in seawater and freshwater,feeds on crustaceans,aquatic insects and occasionally *** existence of digestive enzyme in viscera to act on chitinous exoskeleton of the prey is of *** this study,a chitinase was purified to homogeneity using ammonium sulfate precipitation,DEAE-Sephacel ion exchange,Sephacryl S-200 HR and Superdex 200 gel filtration *** purified protein presents a molecular mass of 58 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE)and results in a single band on native *** to peptide mass fingerprinting,two peptides containing a total of 20 amino acid residues,were 95%identical to a chitinase from yellow perch(Perca flavescens)and 100%identical to the chitinase from greater amberjack(Seriola dumerili).The purified chitinase showed optimum activity at pH 6.0,and was stable at acidic conditions and temperature below 55℃.The enzymatic activity was quite stable in the presence of NaCl,even at 1 mol/*** chitinase was capable of degrading chitosan into low molecular mass chitooligosaccharides(COS)with sizes in a range of 200-700 Da,and the circular dichroism profile of the COS greatly differed from native ***-length cDNA encoding the present chitinase was cloned and the transcript levels of chitinase in various tissues were determined by quantitative real-time *** results showed that the transcript level of chitinase was highest in esophagus and hepatopancreas.