Enzyme co-localisation and aggregation for enhanced metabolic activity for commodity chemicals

石油化工商品的生物替代品是聚合物、涂料和表面活性剂等各种工业应用所需的投入，预计未来十年将占全球产量的2%至25%。国际化学品市场目前价值约4万亿美元，生物生产具有环境可持续性、更低的生产成本和免受油价波动影响等优势。代谢工程和合成生物学等方法可以用来使生物产品和过程更有效率和成本竞争力，并正在推动化学工业的创新。在这项应用中，我们概述了一种方法，该方法通过开发细胞内的酶-聚集体，为生物商品化生产一种名为1，3-丁二醇的化学物质提供了显著的阶梯变化的潜力。我们将把这项技术应用于可以依靠气体废气生存的特定细菌--乙酸菌。该项目结合了两项现有的和受保护的技术(来自肯特大学和ZuvaSyntha的背景IP)，以例证在一种具有商业重要性的菌株中使用酶聚集体，该菌株能够合成商品化学品，并在一家合资企业下产生可能的新的前景IP。细菌微室衍生标签(如P18和D18)的使用将被用来为微胶囊中多酶途径的共定位提供细胞内支架，从而有机会增加通量和催化效率，并保护敏感的细胞内特征，使其免受细胞内潜在有毒的含氧官能化中间体的影响。ZuvaSyntha用于将乙酸转化为1，3-丁二醇的新代谢途径已被开发用于厌氧微生物，并已被计算代谢模型(牛津布鲁克斯大学David Fell教授)证明能够有效地将合成气转化为BD。通过将P18或D18结合到它们的N-末端来产生代谢酶，蛋白质将组装成一个酶聚集体，代表一个内部的细胞生物反应器。这种方法将产生具有显著增强的代谢功能的微生物细胞工厂。该项目的结果将确定蛋白质支架和聚集体在多大程度上可以用于乙酸菌，以增强其代谢能力，以生产通过有毒中间体产生的代谢物。这项研究将揭示这些不同的聚集体可以被重新设计和重新设计以合成重要商品的范围。这样，研究将通过加强与生产重要平台化学品相关的特定代谢过程，提供关于支架在合成气体发酵中应用潜力的初步数据。这项研究还将有助于确定合成生物学的一个重要领域，允许开发支架，并解决内部生物反应器在生物学中的基本优势。在这方面，生物技术工业应用领域的人，特别是对代谢工程感兴趣的人，将普遍对这项研究感兴趣。更具体地说，它还将为生产1，3-丁二醇形式的重要商品化学品提供一个非常先进的系统。

Bio-based alternatives to petrochemical commodities, which are required as inputs into a variety of industrial applications such as polymers, coatings and surfactants, are predicted to rise from 2% to 25% of global production over the next ten years. The international chemical market is currently worth around $4 trillion with bio-production offering advantages such as environmental sustainability, cheaper production costs and protection from oil-price volatility. Approaches such as metabolic engineering and synthetic biology can be used to make bio-products and processes more efficient and cost competitive, and are fueling innovation in the chemical industry. In this application we outline a method that offers the potential for a significant step-change in bio-commodity production of a chemical called 1,3-butanediol through the development of intracellular enzyme-aggregates. We will apply this technology to specific bacteria called acetogens that can live on gaseous exhaust fumes.The project brings together 2 existing and protected technologies (background IP from the University of Kent and ZuvaSyntha) to exemplify the use of enzyme-aggregates in a commercially important strain capable of synthesising commodity chemicals and generate possible novel foreground IP under a joint venture. The use of bacterial microcompartment-derived tags (such as P18 and D18) will be used to provide an intracellular scaffold for the co-localisation of multi-enzyme pathways in microrgansms offering the opportunity to increase flux and catalytic efficiency and protect sensitive intracellular features from the potentially toxic oxyfunctionalised intermediates that arise within the cell. ZuvaSyntha's novel metabolic pathway for the transformation of acetic acid into 1,3-butanediol has been developed for use in anaerobic microorganisms and has been shown by computational metabolic modelling (Prof. David Fell, Oxford Brookes University) to be capable of efficiently converting syngas to BD. By producing the metabolic enzymes with P18 or D18 incorporated into their N-terminii the protein will assemble into an enzyme-aggregate that will represent an internal cellular bioreactor. Such an approach will generate microbial cell factories with significantly enhanced metabolic function.The results of this project will determine to what extent protein scaffolds and aggregates can be used in acetogens to enhance their metabolic capability for the production of metabolites that are generated via toxic intermediates. The research will reveal the scope to which these various aggregates can be redesigned and re-engineered for the synthesis of important commodities. In so doing the research will provide primary data on the potential that scaffolds have for application in syngas fermentation, through the enhancement of specific metabolic process that are associated with the production of important platform chemicals. The research will also help define an important area of synthetic biology, permitting the exploitation of scaffolds and addressing the basic advantages of internal bioreactors in biology. In this respect, the research will be of general interest to those in applied areas of the biotechnology industry, especially those interested in metabolic engineering. More specifically it will also provide a highly advanced system for the generation of an important commodity chemical in the form of 1,3-butanediol.