2. Academic Validation
  3. N-glycosylation as an effective strategy to enhance characteristics of Rhizomucor miehei lipase for biodiesel production

N-glycosylation as an effective strategy to enhance characteristics of Rhizomucor miehei lipase for biodiesel production

  • Enzyme Microb Technol. 2022 Oct:160:110072. doi: 10.1016/j.enzmictec.2022.110072.
Miao Tian 1,  Zhiyuan Wang 2,  Junying Fu 2,  Pengmei Lv 2,  Cuiyi Liang 2,  Zhibing Li 2,  LingMei Yang 2,  Tao Liu 3,  Ming Li 4,  Wen Luo 5
Affiliations

Affiliations

  • 1 Key Laboratory of Renewable Energy, Guangzhou Institute of Energy Conversion, Chinese Academy of Sciences, Guangzhou 510640, People's Republic of China; University of Chinese Academy of Sciences, Beijing 100049, People's Republic of China.
  • 2 Key Laboratory of Renewable Energy, Guangzhou Institute of Energy Conversion, Chinese Academy of Sciences, Guangzhou 510640, People's Republic of China.
  • 3 Guangdong Provincial Key Laboratory of Pharmaceutical Bioactive Substances, Guangdong Pharmaceutical University, Guangzhou 510006, People's Republic of China.
  • 4 Key Laboratory of Renewable Energy, Guangzhou Institute of Energy Conversion, Chinese Academy of Sciences, Guangzhou 510640, People's Republic of China; Guangdong Provincial Key Laboratory of New and Renewable Energy Research and Development, Guangzhou 510640, People's Republic of China.
  • 5 Key Laboratory of Renewable Energy, Guangzhou Institute of Energy Conversion, Chinese Academy of Sciences, Guangzhou 510640, People's Republic of China. Electronic address: luowen@ms.giec.ac.cn.
PMID: 35689964 DOI: 10.1016/j.enzmictec.2022.110072
Abstract

The construction of methanol-resistant lipases with high catalytic activity is world-shattering for biodiesel production. A semi-rational method has been constructed to enhance the properties of Rhizomucor miehei Lipase with propeptide (ProRML) by introducing N-glycosylation sites in the Loop structure. The Enzyme activities of the mutants N288 (1448.89 ± 68.64 U/mg) and N142 (1073.68 ± 33.87 U/mg) increased to 56.09 and 41.56 times relative to that of wild type ProRML (WT, 25.83 ± 0.73 U/mg), respectively. After incubation in 50 % methanol for 2.5 h, the residual activities of N314 and N174-1 were 95 % and 85%, which were higher than the WT (27 %). Additionally, the biodiesel yield of all mutants was increased after a one-time addition of methanol for 24 h. Among them, N288 increased the quantity of biodiesel from colza oil from 9.49 % to 88 %, and N314 increased the amount of biodiesel from waste soybean oil from 8.44% to 70%. This study provides an effective method to enhance the properties of Lipase and improve its application potential in biodiesel production.

Keywords

Biodiesel; Loop structure; Methanol tolerance; N-glycosylation; Rhizomucor miehei lipase.

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