Refining Oxidative Enzyme Systems from Talented Microorganisms for Industrial Biocatalysis.

从用于工业生物催化的天才微生物中精炼氧化酶系统。

• 批准号: BB/N010523/1
• 负责人: John Ward
• 金额: $ 15.74万
• 依托单位: University College London
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2016
• 资助国家: 英国
• 起止时间: 2016 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FN010523%2F1
• 关键词: Refining; Oxidative; Enzyme; Systems; Talented

Microorganisms produce enzymes useful for a wide range of industrial applications but one particularly challenging area is the production of oxidized metabolites of drugs that are being developed and also the environmental fate of metabolites of agrochemicals. The former are produced in humans by liver enzymes and their properties need to be understood, and their presence monitored in clinical trials. The latter are produced in the environment and their properties again need to be understood as they can persist for extended periods. Both metabolite categories can be very difficult to synthesise chemically. Human drug metabolites can be produced using mammalian tissue preparations, and agrochemical metabolites in environmental models, but producing either in the quantities required during new product development can be challenging. Microbial systems offer a very useful alternative. Hypha Discovery has assembled a panel of wild-type bacteria that has proved highly effective in producing target oxidized metabolites for a set of >100 industrially-relevant small organic molecules by whole-cell microbial culture biotransformation, and shows significantly better performance compared to the current commercially available genetically-engineered microbial enzyme preparations which are based on a single type of enzyme. The Hypha Discovery panel has also proved effective in making new derivatives of early-stage pharmaceutical lead compounds with improved properties, in particular solubility, which is important for drug bioavailability. This whole-culture biotransformation approach has limitations, however, with regard to the speed of production of the target metabolite and the scalability of its production to multi-gramme and, eventually, kilogramme quantities. This project aims to address the challenge by identifying the genes encoding individual enzymes responsible for producing oxidized metabolites in the six most talented bacteria from Hypha's organism panel, cloning and introducing these genes into well-characterised host bacteria that can be grown in laboratory cultures under standard conditions at small- or large-scale. This work will be done in collaboration with Professor John Ward's group at University College, London, who have pioneered appropriate processes. The work will involve sequencing the whole genomes of these organisms, identifying the sequences for the enzymes of interest, and undertaking the cloning required to produce genetically-engineered derivatives of the host strains expressing the enzymes of interest along with co-factors required for their full functional activity, with scale-up potential.This new collaboration hopes to deliver a significant advance in the application of oxidative industrial biotechnology, and builds on the previous, and highly complementary, experience of both partners. It is an example of UK academic and industrial institutions providing mutual support that should eventually be of benefit to the pharmaceutical and agrochemical industries, and their customers, worldwide.

微生物产生的酶对广泛的工业应用有用，但一个特别具有挑战性的领域是生产正在开发的药物的氧化代谢产物，也是农业化学代谢物的环境命运。前者是由肝酶在人类中生产的，需要了解其特性，并且在临床试验中监测了它们的存在。后者是在环境中产生的，由于它们可以长时间持续存在，因此需要再次理解其特性。两种代谢产物类别都很难化学合成。可以使用哺乳动物组织制剂和环境模型中的农化代谢产生人类药物代谢产物，但是在新产品开发过程中所需的数量生产可能具有挑战性。微生物系统提供了非常有用的替代方案。菌丝发现已经组装了一组野生型细菌，该小组已被证明在生产目标氧化代谢物方面非常有效，用于一组> 100个> 100个> 100个与工业相关的小有机分子，通过全细胞微生物培养生物转化，并且与当前的遗传性微生物相比，与当前的遗传学构造相比，具有明显的更好的性能。菌丝发现面板也已被证明有效地制造具有改善性能的早期药物铅化合物的新衍生物，特别是溶解度，这对于药物生物利用度很重要。但是，这种全培养生物转化方法具有局限性，但是，对于靶代谢产物的生产速度以及其生产对多弹性的可伸缩性以及最终的千克量。该项目旨在通过确定编码单个酶的基因来应对挑战，这些酶在Hypha的有机体面板的六种最有才华的细菌中产生氧化代谢物，将这些基因的克隆和引入这些基因引入良好的宿主细菌中，这些细菌可以在小型或大型规模的标准条件下在实验室培养物中种植，这些基因可以在实验室中种植。这项工作将与伦敦大学学院的约翰·沃德（John Ward）的小组合作完成，后者开创了适当的流程。这项工作将涉及对这些生物的整个基因组进行测序，确定感兴趣酶的序列，并进行宿主菌株的基因工程衍生物所需的克隆，以表达感兴趣的酶的遗传工程衍生物，表达所需的副因素以及具有全部功能性的构建功能，以实现型号，以实现大量的氧化能力。以前的，高度互补的，双方的经验。这是英国学术和工业机构提供相互支持的一个例子，最终应该对制药和农业化学行业及其客户有利。