Enhanced activity and stability of Rhizomucor miehei lipase by mutating N-linked glycosylation site and its application in biodiesel production

doi:10.1016/j.fuel.2021.121514

GIEC OpenIR

	Enhanced activity and stability of Rhizomucor miehei lipase by mutating N-linked glycosylation site and its application in biodiesel production
	Tian, Miao 1,2; Fu, Junying 1; Wang, Zhiyuan 1; Miao, Changlin 1; Lv, Pengmei1 ; He, Dong 3; Li, Zhibing1 ; Liu, Tao 4; Li, Ming 1; Luo, Wen1
	2021-11-15
发表期刊	FUEL
ISSN	0016-2361
卷号	304 页码:11
通讯作者	Lv, Pengmei(lvpm@ms.giec.ac.cn) ; Luo, Wen(luowen@ms.giec.ac.cn)
摘要	Engineering a lipase with high activity, thermostability, and methanol resistance is of great significance for biodiesel production. To this end, a semi-rational evolutionary method involving site-directed saturation mutagenesis at the N-linked glycosylation site of Rhizomucor miehei lipase has been reported. The enzyme activity of all mutants was improved, particularly N9P, the hydrolytic activity of which was 22 times that of the wild-type Rhizomucor miehei lipase with propeptide. Thermostability and tolerance of all the mutants in 30% methanol were improved, except for N9P. Upon mutation, when the asparagine (N) residue at position 59 was substituted with the charged basic amino acids histidine and lysine, the mutants N59H and N59K were obtained, respectively, both of which showed better performance than the other mutants. During biotransformation of colza oil to biodiesel with one-shot addition of methanol, N59H yielded 87.83% of fatty acid methyl esters in 24 h, a yield significantly higher than obtained using non-glycosylated N9A/N59A (9.49%). Owing to its excellent methanol tolerance and productivity, the genetically engineered mutant N59H has excellent potential for commercial onepot biodiesel production.
关键词	Rhizomucor miehei lipase N-linked glycosylation site Thermostability Methanol tolerance Biodiesel
DOI	10.1016/j.fuel.2021.121514
关键词[WOS]	PICHIA-PASTORIS ; MUTAGENESIS ; PROPEPTIDE ; RESIDUES ; OIL
收录类别	SCI
语种	英语
资助项目	National Natural Science Foundation of China[51903236,51806225] ; Natural Science Foundation of Guangdong Province of China[2020A1515010513] ; Natural Science Foundation of Guangdong Province of China[2021A1515010325]
WOS研究方向	Energy & Fuels ; Engineering
项目资助者	National Natural Science Foundation of China ; Natural Science Foundation of Guangdong Province of China
WOS类目	Energy & Fuels ; Engineering, Chemical
WOS记录号	WOS:000691213500001
出版者	ELSEVIER SCI LTD
引用统计	被引频次：18[WOS] [WOS记录] [WOS相关记录]
文献类型	期刊论文
条目标识符	http://ir.giec.ac.cn/handle/344007/33840
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.Chinese Acad Sci, Guangzhou Inst Energy Convers, Key Lab Renewable Energy, Guangzhou 510640, Peoples R China 2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 3.Guangdong Univ Technol, Guangzhou 510090, Peoples R China 4.Guangdong Pharmaceut Univ, Guangdong Prov Key Lab Pharmaceut Bioact Subst, Guangzhou 510006, Peoples R China
第一作者单位	中国科学院广州能源研究所
推荐引用方式 GB/T 7714	Tian, Miao,Fu, Junying,Wang, Zhiyuan,et al. Enhanced activity and stability of Rhizomucor miehei lipase by mutating N-linked glycosylation site and its application in biodiesel production[J]. FUEL,2021,304:11.
APA	Tian, Miao.,Fu, Junying.,Wang, Zhiyuan.,Miao, Changlin.,Lv, Pengmei.,...&Luo, Wen.(2021).Enhanced activity and stability of Rhizomucor miehei lipase by mutating N-linked glycosylation site and its application in biodiesel production.FUEL,304,11.
MLA	Tian, Miao,et al."Enhanced activity and stability of Rhizomucor miehei lipase by mutating N-linked glycosylation site and its application in biodiesel production".FUEL 304(2021):11.