基于蛋白质组学的菲律宾蛤仔凝集素MCL-T抑制腐败希瓦氏菌机理研究  

Inhibitory mechanism of lectin(MCL-T)against Shewanella putrefaciens in Manila clam Ruditapes philippinarum by proteomics approach

作　　者：杨晴晴 金祎雯 李伟 赵菲 孔亮[2] 佟长青 YANG Qingqing;JIN Yiwen;LI Wei;ZHAO Fei;KONG Liang;TONG Changqing(College of Food Science and Engineering,Dalian Ocean University,Dalian 116023,China;College of Marine Science and Environment,Dalian Ocean University,Dalian 116023,China)

机构地区：[1]大连海洋大学食品科学与工程学院,辽宁大连116023 [2]大连海洋大学海洋科学与环境学院,辽宁大连116023

出　　处：《大连海洋大学学报》2021年第4期653-660,共8页Journal of Dalian Ocean University

基　　金：国家自然科学基金面上项目(31571916)。

摘　　要：为探究菲律宾蛤仔Ruditapes philippinarum凝集素(MCL-T)诱导产生及其抑制腐败希瓦氏菌Shewanella putrefaciens的作用机制,以腐败希瓦氏菌刺激菲律宾蛤仔,并利用固相吸附分析MCL-T与腐败希瓦氏菌外膜蛋白(outer membrane proteins,OMP)的相互作用,利用非标记(Label-free)定量蛋白质组方法对MCL-T作用于腐败希瓦氏菌的蛋白质组进行分析。结果表明:注射比浸泡方式产生更多的MCL-T;MCL-T与腐败希瓦氏菌外膜蛋白结合具有浓度依赖性;MCL-T作用于腐败希瓦氏菌后,有74个蛋白质表达量变化显著,其中,16个蛋白质表达量上调,58个蛋白质表达量下调,这些差异表达蛋白质主要参与氨基酸生物合成途径、丙酮酸代谢途径、柠檬酸循环途径、氨酰-tRNA生物合成途径和碳代谢途径。研究表明,MCL-T是通过影响腐败希瓦氏菌体内氨基酸合成、丙酮酸代谢和柠檬酸循环等实现其抑菌作用。In order to investigate the mechanism of lectin(MCL-T)secretion of Manila clam Ruditapes philippinarum induced by Shewanella putrefaciens and inhibitory effect of the lectin against S.putrefaciens,freeze-dried powder of the MCL-T was in the sea water of Manila clam with body mass of(8±1)g injected with S.putrefaciens at feed muscles at a dose of 0.5,1.0,1.5,2.0,and 2.5 OD 600 nm and reared in clean 300 mL enamel trays of each 20 cm×25 cm×4.5 cm at water temperature of(22±3)℃in 12,24 h,and 36 h by solid phase assay analyzing the reaction of MCL-T with S.putrefaciens outer membrane proteins(OMP),immersion of Manila clam in group 7 and without any treatment of the clam in group 1.OD 600 nm values were measured in diluted sterile TSB culture medium of S.putrefaciens containing freeze-dried powder of the MCL-T at a dose of 0,0.01,0.1,and 1.0 mg/L for 24 h to detect inhibitory effect of MCL-T on S.putrefaciens.The expression of the proteome in S.putrefaciens with MCL-T was analyzed by label-free quantitative proteomics.The results showed that there was more yield of MCL-T in the injection groups than that in the immersion group,with concentration-dependence of MCL-T binding to S.putrefaciens OMP.A total of 74 differential proteins were identified in expression profiles of S.putrefaciens with MCL-T compared with without MCL-T.The expression of 16 proteins was found to be up-regulated and 58 proteins down-regulated.These differentially expressed proteins mainly involved in the amino acid biosynthetic pathway,pyruvate metabolism,citric acid cycle,aminoacyl-tRNA biosynthesis pathway and carbon metabolism pathway.Therefore,MCL-T inhibited S.putrefaciens by disturbing amino acid biosynthesis,pyruvate metabolism and citric acid cycle.

关 键 词：菲律宾蛤仔 凝集素 腐败希瓦氏菌 固相吸附 蛋白质组学 

分 类 号：S917.4[农业科学—水产科学] TS254.1[轻工技术与工程—水产品加工及贮藏工程]