The Key Laboratory of Biotechnology for Medicinal Plant of Jiangsu Province, School of Life Sciences, Jiangsu Normal University, Xuzhou, Jiangsu Province 221116, China.
J Agric Food Chem. 2024 Jul 31;72(30):16835-16847. doi: 10.1021/acs.jafc.4c02360. Epub 2024 Jul 19.
Few reports exist on one-step enzymatic methods for the simultaneous production of biodiesel and eicosapentaenoic acid ethyl ester (EPA-EE), a high-value pharmaceutical compound. This study aimed to efficiently express lipase (pRML) in X-33 via propeptide mutation and high-copy strain screening. The mutated enzyme was then used to simultaneously catalyze the production of both biodiesel and EPA-EE. The P46N mutation in the propeptide (P46N-pRML) significantly boosted its production, with the four-copy strain increasing enzyme yield by 3.7-fold, reaching 3425 U/mL. Meanwhile, its optimal temperature increased to 45-50 °C, pH expanded to 7.0-8.0, specific activity doubled, reduced to one-third, and / increased 7-fold. Notably, P46N-pRML efficiently converts oil's eicosapentaenoic acid (EPA). Under optimal conditions, it achieves up to 93% biodiesel and 92% EPA-EE yields in 9 h. Our study introduces a novel, efficient one-step green method to produce both biodiesel and EPA-EE using this advanced enzyme.
关于一步酶法同时生产生物柴油和高价值药物化合物二十碳五烯酸乙酯（EPA-EE）的报道很少。本研究旨在通过前肽突变和高拷贝菌株筛选，高效表达脂肪酶（pRML）。然后，用突变酶同时催化生物柴油和 EPA-EE 的生产。前肽中的 P46N 突变（P46N-pRML）显著提高了其产量，四拷贝菌株使酶产量增加了 3.7 倍，达到 3425 U/mL。同时，其最适温度升高至 45-50°C，pH 值扩大至 7.0-8.0，比活提高了一倍，半衰期缩短至三分之一，半衰期延长了 7 倍。值得注意的是，P46N-pRML 能有效地转化鱼油中的二十碳五烯酸（EPA）。在最佳条件下，它在 9 小时内可达到高达 93%的生物柴油和 92%的 EPA-EE 产率。本研究采用这种先进的酶，介绍了一种生产生物柴油和 EPA-EE 的新型、高效的一步绿色方法。
