A superior lipase for cheese production: A precisely medium-chain selective lipase from Pleurotus citrinopileatus expressed in Pichia pastoris

文献类型： 外文期刊

第一作者： Wang, Zhulin

作者： Wang, Zhulin;Zhang, Wanqing;Liu, Ziwei;Wang, Jianan;Li, Qian;Li, Binglin;Wang, Zhulin;Liu, Shujun;Zhang, Wanqing;Xin, Fengjiao;Wang, Zhulin;Liu, Shujun;Zhang, Wanqing;Xin, Fengjiao;Li, Binglin;Henrich, Lea;Gand, Martin;Zhang, Tiantian

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关键词： Lipase; Medium-chain; Substrate selectivity; Cheese flavor; Molecular dynamics

期刊名称：FOOD CHEMISTRY （ 影响因子：9.8； 五年影响因子：9.7 ）

ISSN： 0308-8146

年卷期： 2025 年 492 卷

页码：

收录情况： SCI

摘要： The lipase derived from Pleurotus citrinopileatus (PCI-Lip) shows great potential for cheese production but could not achieve high-level expression in E. coli. Herein, we successfully expressed PCI-Lip extracellularly in Pichia pastoris X-33 (PpPCI-Lip), which exhibited a 472-fold increase in activity (kcat/Km = 5.99 x 105 M- 1.S- 1) against p-nitrophenyl octanoate compared to E. coli. Scaled-up fermentation in a 5-L fermentor achieved a high volumetric activity of 5.94 x 105 U.g- 1, 3.17 times higher than shake fermentation. PpPCI-Lip demonstrated higher specificity for medium-chain fatty acids than commercial Palatase 20,000 L, releasing caprylic acid, capric acid, and lauric acid to enhance creamy flavors in cheese, as confirmed by GC-MS. Molecular dynamics simulations revealed that the lid region (Cys65-Ser98) governs substrate entry, enabling high hydrolysis selectivity for medium-chain triglycerides (C10:0) due to lower steric hindrance, while long-chain triglycerides (C18:0, C18:1) showed reduced efficiency due to steric resistance and aggregation effects.

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