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

	Enhanced activity and stability of Rhizomucor miehei lipase by mutating N-linked glycosylation site and its application in biodiesel production
	Tian, Miao 2,3; Fu, Junying 3; Wang, Zhiyuan 3; Miao, Changlin 3; Lv, Pengmei 3; He, Dong 4; Li, Zhibing 3; Liu, Tao 1; Li, Ming 3; Luo, Wen 3
刊名	FUEL
	2021-11-15
卷号	304 页码:11
关键词	Rhizomucor miehei lipase N-linked glycosylation site Thermostability Methanol tolerance Biodiesel
ISSN号	0016-2361
DOI	10.1016/j.fuel.2021.121514
通讯作者	Lv, Pengmei(lvpm@ms.giec.ac.cn) ; Luo, Wen(luowen@ms.giec.ac.cn)
英文摘要	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.
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关键词	PICHIA-PASTORIS ; MUTAGENESIS ; PROPEPTIDE ; RESIDUES ; OIL
WOS研究方向	Energy & Fuels ; Engineering
语种	英语
出版者	ELSEVIER SCI LTD
WOS记录号	WOS:000691213500001
资助机构	National Natural Science Foundation of China ; Natural Science Foundation of Guangdong Province of China
内容类型	期刊论文
源URL	[http://ir.giec.ac.cn/handle/344007/33839]
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.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 3.Chinese Acad Sci, Guangzhou Inst Energy Convers, Key Lab Renewable Energy, Guangzhou 510640, Peoples R China 4.Guangdong Univ Technol, Guangzhou 510090, Peoples R China
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.