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

Acta Oceanologica Sinica 2021年第6期40卷 50-60页

作者： Yulei Chen Zhipeng Tao Minghui Zhang Lechang Sun Guangming Liu Minjie Cao College of Food and Biological Engineering Jimei UniversityXiamen 361021China Fujian Collaborative Innovation Center for Exploitation and Utilization of Marine Biological Resources Xiamen 361102China

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.

关键词： bostrychus sinensis chitinase purification characterization molecular cloning chitosan

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