Zhang Yao, Sun Yan, Liu Zhuo, Leng Jiajun, Liu Qing, Song Yuanda
Food Bioengineering and Technology Laboratory, Department of Food Science and Nutrition, College of Culture and Tourism, University of Jinan, 13 Shungeng Road, Jinan, 250022, People's Republic of China.
Colin Ratledge Center for Microbial Lipids, School of Agricultural Engineering and Food Science, Shandong University of Technology, 266 Xincun West Road, Zibo, 255000, People's Republic of China.
Sci Rep. 2025 Mar 29;15(1):10871. doi: 10.1038/s41598-025-93938-x.
Microbial lipases could be used to hydrolyze or recombine fats and oils, and had great applications in food processing, bioenergy and chemical industry. Mucor circinelloides was an important gamma-linolenic acid producing strain, and its genome was predicted to contain a large number of genes encoding lipases, the key enzymes in lipid metabolism. In the present study, a potential lipase WJ_23 from Mucor circinelloides WJ11 was cloned for the first time and heterologously expressed and purified to homogeneity in Pichia pastoris. By SDS-PAGE analysis, the molecular weight of the recombinant lipase was estimated to be ~ 34 kDa. The optimal temperature and pH of the recombinant lipase were 50 °C and 9.0, respectively. The recombinant lipase had good thermal stability at 50 °C with a broad pH stability from 6.0 to 11.0. After incubation at 37 °C for 24 h, the activity of the recombinant lipase remained over 95% between pH 7.0 and 9.0. The recombinant lipase possessed a preference for the long chain substrates. Using p-NPP as substrate, the measured kinetic parameters V and K were 94.34 U/mg and 6.37 mmoL/L, respectively. In addition, the activity of the recombinant lipase was not affected obviously by various metal ions, and it exhibited certain stability and tolerance towards several kinds of organic solvents. This research might provide a basis for the further industrial application of recombinant M. circinelloides lipase.
微生物脂肪酶可用于水解或重组油脂，在食品加工、生物能源和化工领域有广泛应用。卷枝毛霉是一种重要的γ-亚麻酸生产菌株，其基因组预计包含大量编码脂肪酶的基因，脂肪酶是脂质代谢中的关键酶。在本研究中，首次从卷枝毛霉WJ11中克隆了一种潜在的脂肪酶WJ_23，并在毕赤酵母中进行了异源表达和纯化，使其达到均一性。通过SDS-PAGE分析，重组脂肪酶的分子量估计约为34 kDa。重组脂肪酶的最适温度和pH分别为50℃和9.0。重组脂肪酶在50℃时具有良好的热稳定性，在pH 6.0至11.0范围内具有较宽的pH稳定性。在37℃孵育24小时后，重组脂肪酶在pH 7.0至9.0之间的活性保持在95%以上。重组脂肪酶对长链底物具有偏好性。以对硝基苯酚棕榈酸酯（p-NPP）为底物，测得的动力学参数V和Km分别为94.34 U/mg和6.37 mmol/L。此外，重组脂肪酶的活性不受各种金属离子的明显影响，并且对几种有机溶剂表现出一定的稳定性和耐受性。本研究可能为重组卷枝毛霉脂肪酶的进一步工业应用提供依据。
