Lipases are a group of enzymes capable of catalyzing the hydrolysis of triacylglycerides into free fatty acids during lipid metabolism. In the presence of short-chain alcohols, some of these enzymes can catalyze the transesterification of plant oils into biodiesel. Biodiesel has recently gained traction to be a green alternative to fossil fuel. Efficient production of biodiesel at an industrial scale is limited by the reusability of the enzyme and its tolerance toward high concentrations of organic solvents. We describe a scalable workflow that integrates web-based tools and automation to identify a group of 114 orthologous bacterial lipases for expression, purification, and characterization using a high-throughput platform at our biofoundry. The activity profile of these enzymes revealed many targets with different substrate specificities and an optimal pH range. Most of these enzymes are thermostable and can tolerate up to 40% methanol (v/v). Among the 114 lipases that were targeted, 22 were found to be able to produce methyl oleate from triolein in the presence of methanol. Our study demonstrates the utility of the workflow to identify lipase candidates for the industrial production of biodiesel.

中文翻译：

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绿色制造的可扩展工作流程：发现用于生物柴油生产的细菌脂肪酶

脂肪酶是一组能够在脂质代谢过程中催化甘油三酯水解为游离脂肪酸的酶。在短链醇的存在下，这些酶中的一些可以催化植物油酯交换为生物柴油。生物柴油最近受到关注，成为化石燃料的绿色替代品。以工业规模有效生产生物柴油受到酶的可重复使用性及其对高浓度有机溶剂的耐受性的限制。我们描述了一个可扩展的工作流程，它集成了基于网络的工具和自动化，以使用我们生物铸造厂的高通量平台识别一组 114 种直系同源细菌脂肪酶，用于表达、纯化和表征。这些酶的活性谱揭示了许多具有不同底物特异性和最佳 pH 范围的目标。大多数这些酶是热稳定的，可以耐受高达 40% 的甲醇 (v/v)。在目标的 114 种脂肪酶中，发现 22 种能够在甲醇存在下从三油精生产油酸甲酯。我们的研究证明了该工作流程可用于识别用于生物柴油工业生产的脂肪酶候选物。