Enhancement of Streptomyces transglutaminase activity and pro-peptide cleavage efficiency by introducing linker peptide in the C-terminus of the pro-peptide

被引:8
作者
Chen, Kangkang [1 ,2 ]
Liu, Song [1 ,2 ]
Wang, Guangsheng [1 ]
Zhang, Dongxu [1 ,3 ]
Du, Guocheng [1 ,4 ]
Chen, Jian [1 ,5 ]
Shi, Zhongping [1 ,2 ]
机构
[1] Jiangnan Univ, Sch Biotechnol, Wuxi 214122, Peoples R China
[2] Jiangnan Univ, Minist Educ, Key Lab Ind Biotechnol, Wuxi, Peoples R China
[3] Jiangnan Univ, State Key Lab Food Sci & Technol, Wuxi, Peoples R China
[4] Jiangnan Univ, Minist Educ, Key Lab Carbohydrate Chem & Biotechnol, Wuxi, Peoples R China
[5] Jiangnan Univ, Natl Engn Lab Cereal Fermentat Technol, Wuxi, Peoples R China
基金
国家高技术研究发展计划(863计划); 中国国家自然科学基金;
关键词
Transglutaminase; Pro-peptide; Protein folding; Specific activity; Linker peptide; MICROBIAL TRANSGLUTAMINASE; RANDOM MUTAGENESIS; PROTEIN; MOBARAENSIS; STREPTOVERTICILLIUM; EXPRESSION; SUBSTRATE; SEQUENCE; REGION;
D O I
10.1007/s10295-012-1221-y
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Streptomyces transglutaminase (TGase) has been widely used in food, pharmaceutical and textile industries. Streptomyces TGase is naturally synthesized as zymogen (pro-TGase), which is then processed to produce active enzyme by removing its N-terminal pro-peptide. Although the pro-peptide is essential for TGase folding and secretion, few studies have been reported on improving the properties of TGase by pro-peptide engineering. In this study, we developed a new approach to improve the properties of TGase based on pro-peptide engineering. When the alpha-helix(37G-42S) in pro-peptide was substituted with three glycines and three alanines respectively, the mutants exhibited higher specific activity and the efficiency of pro-peptide cleavage was enhanced. To further improve the properties of TGase, relevant mutations were constructed by introducing linker peptides in the C-terminus of the pro-peptide. Mutants with GS (GGGGS) and PT (PTPPTTPT) linker peptide exhibited 1.28 fold and 1.5 fold higher specific activity than the wild-type enzyme, respectively. This new method could be used to improve the properties of TGase by pro-peptide modification, which is a promising technology for creating unique TGase with various beneficial properties.
引用
收藏
页码:317 / 325
页数:9
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