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
作者: Zhulin Wang;Shujun Liu;Lea Henrich;Wanqing Zhang;Tiantian Zhang;Ziwei Liu;Jianan Wang;Qian Li;Martin Gand;Fengjiao Xin;Binglin Li
作者机构:
关键词: Cheese flavor;Lipase;Medium-chain;Molecular dynamics;Substrate selectivity
期刊名称: Food Chemistry
ISSN: 0308-8146
年卷期: 2025 年 492 卷
页码:
收录情况: SCIE(2025版) ; ; EI(2025版)
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 × 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 × 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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