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Improved methanol tolerance of Rhizomucor miehei lipase based on N?glycosylation within the α-helix region and its application in biodiesel production
Tian,Miao1,2; Yang,Lingmei1; Wang,Zhiyuan1; Lv,Pengmei1; Fu,Junying1; Miao,Changlin1; Li,Ming1,3; Liu,Tao4; Luo,Wen1
2021-12-15
发表期刊Biotechnology for Biofuels
卷号14期号:1
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AbstractBackgroundLiquid 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 α/β-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.ResultsIn 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 α-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.ConclusionsThese results indicate that optimising N-glycosylation modification in the α-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 α-Helix region N?Glycosylation Methanol tolerance Biodiesel
DOI10.1186/s13068-021-02087-6
语种英语
WOS记录号BMC:10.1186/s13068-021-02087-6
出版者BioMed Central
引用统计
文献类型期刊论文
条目标识符http://ir.giec.ac.cn/handle/344007/35216
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 Academy of Sciences; Key Laboratory of Renewable Energy, Guangzhou Institute of Energy Conversion
2.University of Chinese Academy of Sciences
3.Guangdong Provincial Key Laboratory of New and Renewable Energy Research and Development
4.Guangdong Pharmaceutical University; Guangdong Provincial Key Laboratory of Pharmaceutical Bioactive Substances
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 α-helix region and its application in biodiesel production[J]. Biotechnology for Biofuels,2021,14(1).
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 α-helix region and its application in biodiesel production.Biotechnology for Biofuels,14(1).
MLA Tian,Miao,et al."Improved methanol tolerance of Rhizomucor miehei lipase based on N?glycosylation within the α-helix region and its application in biodiesel production".Biotechnology for Biofuels 14.1(2021).
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