Improved methanol tolerance of Rhizomucor miehei lipase based on N-glycosylation within the alpha-helix region and its application in biodiesel production

doi:10.1186/s13068-021-02087-6

GIEC OpenIR

	Improved methanol tolerance of Rhizomucor miehei lipase based on N-glycosylation within the alpha-helix region and its application in biodiesel production
	Tian, Miao 1,2; Yang, Lingmei1 ; Wang, Zhiyuan 1; Lv, Pengmei1 ; Fu, Junying 1; Miao, Changlin 1; Li, Ming 1,3; Liu, Tao 4; Luo, Wen1
	2021-12-15
发表期刊	BIOTECHNOLOGY FOR BIOFUELS
卷号	14 期号:1 页码:10
通讯作者	Lv, Pengmei(lvpm@ms.giec.ac.cn) ; Liu, Tao(tliu@foxmail.com) ; Luo, Wen(luowen@ms.giec.ac.cn)
摘要	Background: Liquid lipases are widely used to convert oil into biodiesel. Methanol-resistant lipases with high catalytic activity are the first choice for practical production. Rhizomucor miehei lipase (RML) is a single-chain alpha/beta-type protein that is widely used in biodiesel preparation. Improving the catalytic activity and methanol tolerance of RML is necessary to realise the industrial production of biodiesel. Results: In this study, a semi-rational design method was used to optimise the catalytic activity and methanol tolerance of ProRML. After N-glycosylation modification of the alpha-helix of the mature peptide in ProRML, the resulting mutants N218, N93, N115, N260, and N183 increased enzyme activity by 66.81, 13.54, 10.33, 3.69, and 2.39 times than that of WT, respectively. The residual activities of N218 and N260 were 88.78% and 86.08% after incubation in 50% methanol for 2.5 h, respectively. In addition, the biodiesel yield of all mutants was improved when methanol was added once and reacted for 24 h with colza oil as the raw material. N260 and N218 increased the biodiesel yield from 9.49% to 88.75% and 90.46%, respectively. Conclusions: These results indicate that optimising N-glycosylation modification in the alpha-helix structure is an effective strategy for improving the performance of ProRML. This study provides an effective approach to improve the design of the enzyme and the properties of lipase mutants, thereby rendering them suitable for industrial biomass conversion.
关键词	Rhizomucor miehei lipase alpha-Helix region N-Glycosylation Methanol tolerance Biodiesel
DOI	10.1186/s13068-021-02087-6
关键词[WOS]	RECOMBINANT ELASTASE ; ENHANCED EXPRESSION ; YEAST ; THERMOSTABILITY ; PROPEPTIDE ; STABILITY ; RESIDUES ; SITES
收录类别	SCI
语种	英语
资助项目	Natural Science Foundation of Guangdong Province of China[2020A1515010513] ; Natural Science Foundation of Guangdong Province of China[2021A1515010325] ; National Natural Science Foundation of China[51903236] ; National Natural Science Foundation of China[51806225] ; Guangdong Provincial Key Laboratory of New and Renewable Energy Research and Development[E039kf0701]
WOS研究方向	Biotechnology & Applied Microbiology ; Energy & Fuels
项目资助者	Natural Science Foundation of Guangdong Province of China ; National Natural Science Foundation of China ; Guangdong Provincial Key Laboratory of New and Renewable Energy Research and Development
WOS类目	Biotechnology & Applied Microbiology ; Energy & Fuels
WOS记录号	WOS:000730561800001
出版者	BMC
引用统计	被引频次：7[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/344007/35525
专题	中国科学院广州能源研究所
通讯作者	Lv, Pengmei; Liu, Tao; 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 Prov Key Lab New & Renew able Energy Re, Guangzhou, 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,Yang, Lingmei,Wang, Zhiyuan,et al. Improved methanol tolerance of Rhizomucor miehei lipase based on N-glycosylation within the alpha-helix region and its application in biodiesel production[J]. BIOTECHNOLOGY FOR BIOFUELS,2021,14(1):10.
APA	Tian, Miao.,Yang, Lingmei.,Wang, Zhiyuan.,Lv, Pengmei.,Fu, Junying.,...&Luo, Wen.(2021).Improved methanol tolerance of Rhizomucor miehei lipase based on N-glycosylation within the alpha-helix region and its application in biodiesel production.BIOTECHNOLOGY FOR BIOFUELS,14(1),10.
MLA	Tian, Miao,et al."Improved methanol tolerance of Rhizomucor miehei lipase based on N-glycosylation within the alpha-helix region and its application in biodiesel production".BIOTECHNOLOGY FOR BIOFUELS 14.1(2021):10.