Xiao Dunchi, Li Xun, Zhang Yu, Wang Fei
Co-Innovation Center for Efficient Processing and Utilization of Forest Resources, College of Chemical Engineering, Nanjing Forestry University, Nanjing, 210037, China.
Jiangsu Provincial Key Lab for the Chemistry and Utilization of Agro-Forest Biomass, Nanjing Forestry University, Nanjing, 210037, China.
Appl Biochem Biotechnol. 2025 Mar;197(3):2035-2050. doi: 10.1007/s12010-024-05095-y. Epub 2024 Dec 7.
In this study, the liquid lipase Yarrowia lipolytica lipase 2 (YLLip2) expressed by Pichia pastoris GS115 was used to produce biodiesel from waste oil. Four signal peptides were compared to express YLLip2 in P. pastoris, among which SP23 exhibited greater secretion performance. In a 1.3-L bioreactor with FM22 medium for 7 days incubation, the maximum YLLip2 activity and total protein content reached 895.44 ± 27.31 U/mL and 3.83 ± 0.31 g/L, respectively. Under the optimal reaction conditions of 30 °C, 20% moisture content, 50 U/g oil of enzyme dosage, and distributed methanol addition, the reaction yield reached 80.99% after 12 h. To further improve the biodiesel yield, some additives were used to assist with YLLip2. The results showed that adding G50 (approximately 1/20 of YLLip2) increased the yield by approximately 90% after 6 h cultivation without changing the enzyme dosage. Compared with previous studies, the reaction time for biodiesel production from waste oil in this study was significantly shortened. This study provides a workable method for converting low-quality feedstocks containing high free fatty acids into biodiesel using a liquid lipase as the catalyst.
在本研究中，利用毕赤酵母GS115表达的解脂耶氏酵母液体脂肪酶2（YLLip2）从废油中生产生物柴油。比较了四种信号肽在毕赤酵母中表达YLLip2的情况，其中SP23表现出更高的分泌性能。在装有FM22培养基的1.3-L生物反应器中培养7天，YLLip2的最大活性和总蛋白含量分别达到895.44±27.31 U/mL和3.83±0.31 g/L。在30℃、水分含量20%、酶用量50 U/g油以及分批添加甲醇的最佳反应条件下，12小时后反应产率达到80.99%。为进一步提高生物柴油产率，使用了一些添加剂辅助YLLip2。结果表明，添加G50（约为YLLip2的1/20）在不改变酶用量的情况下，培养6小时后产率提高了约90%。与之前的研究相比，本研究中利用废油生产生物柴油的反应时间显著缩短。本研究提供了一种可行的方法，即以液体脂肪酶为催化剂，将含有高游离脂肪酸的低质量原料转化为生物柴油。
