苏云金芽胞杆菌分泌蛋白的鉴定及分析

doi:10.13560/j.cnki.biotech.bull.1985.2017.04.022

摘要/Abstract

摘要： 研究苏云金芽胞杆菌（Bacillus thuringiensis,Bt）在生长发育过程中分泌蛋白的类型和含量,尤其是胞外蛋白酶,对于充分了解该菌的杀虫机理、扩大该菌的应用具有重要意义。通过制备Bt_HD73过渡期（transition phase）的分泌蛋白质样品,进行质谱检测并分析其分泌蛋白的种类、含量、功能和信号肽等。结果表明,Bt_HD73在过渡期初期有54种胞外蛋白质,其中酶类最多,占所有分泌蛋白的66.30%,且主要为蛋白水解酶;毒力蛋白次之,占所有分泌蛋白的14.53%。在54种分泌蛋白中,发现27种含有经典Sec型信号肽（Secretory signal peptide,Sec-type SP）,信号肽的长度在24-40个氨基酸之间。因此,Bt_HD73产生分泌蛋白的能力较强,在过渡期可分泌大量酶类,尤其是蛋白水解酶。同时还建立了该菌株的信号肽库,包括27种具有引导外源蛋白分泌潜力的信号肽。

关键词: 苏云金芽胞杆菌, 分泌表达, 分泌蛋白质, 胞外蛋白酶, 信号肽

Abstract: The study on Bacillus thuringiensis（Bt）secretory protein types and their contents,especially the extracellular proteases,is of great significance for fully understanding the insecticidal mechanism and expanding the application of this bacterium. In our research,the secretory protein samples in the transition phase of Bt_HD73 were detected by mass spectrometry,then the types,contents,functions and signal peptides of secretory proteins were analyzed. The results showed that Bt_HD73 secreted 54 kinds of extracellular proteins in transition phase,and most of them were enzymes,accounting for 66.30% of all secretory proteins. Moreover,most of the enzymes were proteases,and the followed was virulence protein,accounting for 14.53% of all secretory proteins. Among the 54 secretory proteins,27 proteins were found to contain the classic Sec-type signal peptides,and the lengths were between 24 and 40 amino acid residues. Therefore,Bt_HD73 has a strong secretory ability,and it can secrete large amounts of enzymes especially proteases in the transition phase. Concurrently,the signal peptide library of this strain was established in this study,including 27 signal peptides with potential for guiding the secretion of heterologous proteins.

Key words: Bacillus thuringiensis, secretory expression, extracellular protease, secretory protein, signal peptide

王智文, 陈海波, 宋福平, 郭淑元. 苏云金芽胞杆菌分泌蛋白的鉴定及分析[J]. 生物技术通报, 2017, 33(4): 169-176.

WANG Zhi-wen, CHEN Hai-bo, SONG Fu-ping, GUO Shu-yuan. Identification and Analysis of Secretory Proteins in Bacillus thuringiensis[J]. Biotechnology Bulletin, 2017, 33(4): 169-176.

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