Engineering new lipases for nutritional applications

在过去的十年中，科学研究报告了与增加体重相关的许多健康益处。 摄入富含omega-3的油。由于这些油在饮食中往往不足，全球对 高质量的营养补充剂为每年数十亿美元的产业提供燃料，并且正在蓬勃发展。世界上大多数供应 是从鱼油中提取的，但目前的提取方法的回收率低。因此，补充 只有适度富含有益的omega-3油，这意味着必须服用更大的剂量。在 特别是，选择性差的催化剂和冗长的反应时间削弱了ω-3油的回收。在 这项协调一致的研究工作，Joelle Pelletier的酶进化实验室将指导 DSM Nutritional Products Canada为生产富含omega-3的油提供更好的生物催化剂。生物催化剂， 或酶，提供了一种消费者安全和环境无害的方法来工业富集omega-3 油.帝斯曼每年使用脂肪酶类天然酶加工数百万公斤鱼油。 尽管长期的尝试，以确定脂肪酶具有更好的性能，他们的ω-3富集率站 上限为60%。为了将omega-3含量提高到70- 80%，我们合作开发了 一个将所需特性工程化到酶中的平台（由Engage资助，2015年4月至10月）。 我们使用计算指导对脂肪酶进行了遗传修饰，并建立了一个筛选 为了以高通量的形式改进omega-3富集，使用机器人自动化来查询特定的 功能.我们目前正准备扩大这一努力，以生产数百种改进的变种， 严格的测试。我们将采用具有最合适性质（包括表达、稳定性、pH）的变体 曲线），用于DSM的反应条件的放大和优化。该项目将产生一个 用于商业用途的改进的生物催化剂，更好地获得高质量的营养补充剂，以及独特的 应用生物催化剂工程的培训机会。

Over the past decade, scientific research has reported on numerous health benefits associated with increased intake of omega-3-enriched oils. Because these oils are often insufficient in the diet, the global demand for high quality supplements fuels a multi-billion $/yr industry, and is booming. The majority of the world supply is extracted from fish oils, yet current extraction processes suffer from low recovery. As a result, supplements are only moderately enriched in the beneficial omega-3 oils, which means larger doses must be taken. In particular, the recovery of omega-3 oils is crippled by poorly selective catalysts and lengthy reaction times. In this concerted research effort, the enzyme evolution laboratory of Joelle Pelletier will guide the engineering of better biocatalysts for production of omega-3-enriched oils at DSM Nutritional Products Canada. Biocatalysts, or enzymes, provide a consumer-safe and environmentally sound approach to industrial enrichment of omega-3 oils. DSM processes millions of kg of fish oil per year using naturally-occurring enzymes from the lipase class. Despite lengthy attempts to identify lipases with better properties, their omega-3 enrichment ratio stands capped at 60%. With the goal of increasing the omega-3 content to 70-80%, we have collaboratively developed a platform to engineer the desired properties into the enzyme (supported by an Engage grant, April-Oct 2015). We genetically modified a lipase enzyme using computational guidance, and established a protocol to screen for improved omega-3 enrichment in a high-throughput format, using robotic automation to query specific features. We are currently poised to scale up this effort, to produce hundreds of improved variants for more rigorous testing. We will take the variant with the most suitable properties (including expression, stability, pH profile) forward for scale-up and optimization of reaction conditions at DSM. This project will yield an improved biocatalyst for commercial use, better access to high quality nutritional supplements, and a unique training opportunity in applied biocatalyst engineering.