Enhanced activity of Rhizomucor miehei lipase by directed saturation mutation of the propeptide

doi:10.1016/j.enzmictec.2021.109870

	Enhanced activity of Rhizomucor miehei lipase by directed saturation mutation of the propeptide
	Tian, Miao 3,4; Huang, Shaowei 2; Wang, Zhiyuan 4; Fu, Junying 4; Lv, Pengmei 4; Miao, Changlin 4; Liu, Tao 1; Yang, Lingmei 4; Luo, Wen 4
刊名	ENZYME AND MICROBIAL TECHNOLOGY
	2021-10-01
卷号	150 页码:8
关键词	Propeptide RML Enzyme activity Directed saturation mutation
ISSN号	0141-0229
DOI	10.1016/j.enzmictec.2021.109870
通讯作者	Lv, Pengmei(lvpm@ms.giec.ac.cn) ; Luo, Wen(luowen@ms.giec.ac.cn)
英文摘要	The propeptide is a short sequence that facilitates protein folding. In this study, four highly active Rhizomucor miehei lipase (RML) mutants were obtained through saturation mutagenesis at three propeptide positions: Ser8, Pro35, and Pro47. The enzyme activities of mutants P35 N, P47 G, P47 N, and S8E/P35S/P47A observed at 40 degrees C, and pH 8.0 were 10.19, 7.53, 6.15, and 8.24 times of that wild-type RML, respectively. The S8E/P35S/ P47A mutant showed good thermostability. After incubation at 40 degrees C for 1 h, 98.98 % of its initial activity remained, whereas wild-type RML retained only 78.76 %. This result indicated that the enhancement of hydrophilicity of 35- and 47- amino-acid residues could promote the interaction between the propeptide and the mature peptide and the enzyme activity and expression level. Highly conserved sites had a more significant impact on enzyme performance than did other sites, similar to the Pro35 and Pro47 mutants showed in this study. This study provides a new idea for protein modification: enzyme performance can be improved through propeptide regulation.
资助项目	National Natural Science Foundation of China[51903236] ; National Natural Science Foundation of China[51806225] ; National Natural Science Foundation of China[51861145103] ; National Key Research and Development Projects[2019YFB1504003]
WOS关键词	INDUSTRIAL-PRODUCTION ; BIODIESEL PRODUCTION ; PICHIA-PASTORIS ; EXPRESSION ; EVOLUTION ; OPTIMIZATION ; MUTAGENESIS ; SUBTILISIN
WOS研究方向	Biotechnology & Applied Microbiology
语种	英语
出版者	ELSEVIER SCIENCE INC
WOS记录号	WOS:000703947600007
资助机构	National Natural Science Foundation of China ; National Key Research and Development Projects
内容类型	期刊论文
源URL	[http://ir.giec.ac.cn/handle/344007/34822]
专题	中国科学院广州能源研究所
通讯作者	Lv, Pengmei; Luo, Wen
作者单位	1.Guangdong Pharmaceut Univ, Guangdong Prov Key Lab Pharmaceut Bioact Subst, Guangzhou 510006, Peoples R China 2.South China Agr Univ, Guangzhou 510642, Peoples R China 3.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 4.Chinese Acad Sci, Guangzhou Inst Energy Convers, Key Lab Renewable Energy, Guangzhou 510640, Peoples R China
推荐引用方式 GB/T 7714	Tian, Miao,Huang, Shaowei,Wang, Zhiyuan,et al. Enhanced activity of Rhizomucor miehei lipase by directed saturation mutation of the propeptide[J]. ENZYME AND MICROBIAL TECHNOLOGY,2021,150:8.
APA	Tian, Miao.,Huang, Shaowei.,Wang, Zhiyuan.,Fu, Junying.,Lv, Pengmei.,...&Luo, Wen.(2021).Enhanced activity of Rhizomucor miehei lipase by directed saturation mutation of the propeptide.ENZYME AND MICROBIAL TECHNOLOGY,150,8.
MLA	Tian, Miao,et al."Enhanced activity of Rhizomucor miehei lipase by directed saturation mutation of the propeptide".ENZYME AND MICROBIAL TECHNOLOGY 150(2021):8.