Alzahrani Areej Ali, Krayem Najeh, Alonazi Mona, Al-Ghamdi Jihan M, Horchani Habib, Ben Bacha Abir
Department of Biochemistry, College of Science, King Saud University, Riyadh, Saudi Arabia.
Laboratory of Biochemistry and Enzymatic Engineering of Lipases, ENIS, University of Sfax, Sfax, Tunisia.
Front Bioeng Biotechnol. 2025 May 16;13:1589087. doi: 10.3389/fbioe.2025.1589087. eCollection 2025.
BACKGROUND/OBJECTIVES: The growing demand for reliable and stable biocatalysts has spurred research into microbial lipases for diverse industrial applications. This study focused on enhancing the production and purification of a lipase from (Lip).
Maximal lipase activity (420 U/mL) was achieved during the stationary phase after 84 h of incubation at 45°C and pH 8.0, using 2% glucose and 2% yeast extract as carbon and nitrogen sources, respectively.
Calcium, olive oil, and Tween, at 1%, significantly enhanced Lip production, highlighting the role of triglycerides and detergents in enzyme induction and substrate emulsification. The purified 50-kDa enzyme displayed maximal activity at 50°C and pH 9.0, with thermal stability between 40°C and 55°C and pH 5.0-10.0. While Lip efficiently hydrolyzed short and medium-chain triglycerides, it exhibited a preference for long-chain substrates, with a maximum reaction rate of 2500 μmol/min/mg and a K value of 6.45 mM toward triolein (C18). Lip also demonstrated remarkable stability in detergent formulations, retaining more than 85% activity in the presence of surfactants, oxidizing agents, boron compounds, and enzyme inhibitors. Additionally, Lip catalyzed the esterification of oleic acid with starch and ethanol to produce starch oleate and ricinoleic acid.
These findings establish Lip as a promising biocatalyst for applications in biocatalysis and detergent formulations, with potential uses in the food, beverage, cosmetic, and pharmaceutical industries.
背景/目的：对可靠且稳定的生物催化剂的需求不断增长，这推动了对用于各种工业应用的微生物脂肪酶的研究。本研究聚焦于提高来自（Lip）的脂肪酶的生产和纯化。
在45°C和pH 8.0下孵育84小时后，使用2%葡萄糖和2%酵母提取物分别作为碳源和氮源，在稳定期达到最大脂肪酶活性（420 U/mL）。
1%的钙、橄榄油和吐温显著提高了Lip的产量，突出了甘油三酯和洗涤剂在酶诱导和底物乳化中的作用。纯化后的50 kDa酶在50°C和pH 9.0时表现出最大活性，热稳定性在40°C至55°C之间，pH值在5.0 - 10.0之间。虽然Lip能有效水解短链和中链甘油三酯，但它对长链底物表现出偏好，对三油酸甘油酯（C18）的最大反应速率为2500 μmol/min/mg，K值为6.45 mM。Lip在洗涤剂配方中也表现出显著的稳定性，在表面活性剂、氧化剂、硼化合物和酶抑制剂存在的情况下保留超过85%的活性。此外，Lip催化油酸与淀粉和乙醇的酯化反应，生成油酸淀粉酯和蓖麻油酸。
这些发现确立了Lip作为一种有前途的生物催化剂，可用于生物催化和洗涤剂配方，在食品、饮料、化妆品和制药行业具有潜在用途。
