Refining Oxidative Enzyme Systems from Talented Microorganisms for Industrial Biocatalysis.

微生物产生对广泛的工业应用有用的酶，但一个特别具有挑战性的领域是正在开发的药物的氧化代谢物的生产，以及农用化学品代谢物的环境命运。前者在人体内由肝酶产生，其性质需要了解，并在临床试验中监测它们的存在。后者是在环境中产生的，需要再次了解它们的性质，因为它们可以长期存在。这两类代谢物都很难用化学方法合成。人类药物代谢物可以用哺乳动物组织制剂生产，农用化学代谢物可以在环境模型中生产，但要生产出新产品开发所需的数量都可能是具有挑战性的。微生物系统提供了一个非常有用的替代方案。Hypha Discovery已经组装了一组野生型细菌，事实证明，通过全细胞微生物培养生物转化，该小组可以高效地为一组与&gt；100工业相关的小有机分子生产目标氧化代谢物，并且与目前商业上可用的基于单一类型酶的转基因微生物酶制剂相比，表现出显著更好的性能。Hypha Discovery小组还被证明有效地制造了早期药物先导化合物的新衍生物，这些化合物具有改善的性质，特别是对药物生物利用度非常重要的溶解性。然而，这种全培养生物转化方法在目标代谢物的生产速度和其生产的可伸缩性方面存在局限性，其生产可以达到多克，最终达到千克量。该项目旨在解决这一挑战，方法是从Hypha的有机体小组中识别出六种最有才华的细菌中负责产生氧化代谢物的个别酶的编码基因，并将这些基因克隆并引入到特征良好的宿主细菌中，这些宿主细菌可以在标准条件下在实验室小规模或大规模培养。这项工作将与伦敦大学学院约翰·沃德教授的团队合作完成，他们开创了适当的过程。这项工作将涉及对这些生物的整个基因组进行测序，确定感兴趣的酶的序列，并进行克隆，以生产表达感兴趣的酶的宿主菌株的基因工程衍生品以及其全部功能活动所需的辅因子，并具有扩大规模的潜力。这一新的合作希望在氧化工业生物技术的应用方面取得重大进展，并建立在双方以前高度互补的经验的基础上。这是英国学术和工业机构相互支持的一个例子，最终应该有利于制药和农化行业及其全球客户。

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.