| Autor: |
Lu, Tong, Qian, Yifan, Zhu, YuQing, Ju, Xiuyun, Dai, Weiwei, Xu, Quanbin, Yang, Qianqian, Li, Shuting, Yuan, Bo, Huang, Jinjin |
| Zdroj: |
Journal of Agricultural and Food Chemistry; July 2024, Vol. 72 Issue: 30 p16835-16847, 13p |
| Abstrakt: |
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 Rhizomucor mieheilipase (pRML) in Pichia pastorisX-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, Kmreduced to one-third, and kcat/Kmincreased 7-fold. Notably, P46N-pRML efficiently converts Nannochloropsisgaditanaoil’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. |
| Databáze: |
Supplemental Index |
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