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

Miao Tian, Zhiyuan Wang, Junying Fu, Pengmei Lv, Cuiyi Liang, Zhibing Li, LingMei Yang, Tao Liu, Ming Li, Wen Luo

Enzyme and microbial technology 2022 Oct

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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. Copyright © 2022 Elsevier Inc. All rights reserved.

Citation

Miao Tian, Zhiyuan Wang, Junying Fu, Pengmei Lv, Cuiyi Liang, Zhibing Li, LingMei Yang, Tao Liu, Ming Li, Wen Luo. N-glycosylation as an effective strategy to enhance characteristics of Rhizomucor miehei lipase for biodiesel production. Enzyme and microbial technology. 2022 Oct;160:110072