Tungsten Biocatalysis - Heavy Metal Enzymes for Sustainable Industrial Biocatalysis

欧洲需要一个可持续的化学工业，只有通过新的突破性技术才能实现。工业生物技术建立在化学制造领域，提供更高效、更具体、更安全和对能源要求较低的生产，但受制于工业用途中有限的酶类。该项目开辟了一类重要的新型含钨酶(W酶)，可催化惊人的化学反应，包括具有挑战性的低氧化还原电位还原反应，但目前无法经济和大规模地获得以满足工业需求。我们需要使用工业主力微生物如大肠杆菌来生产W-酶。然而，我们发现W-辅因子的生物合成是阻碍W-酶在大肠杆菌中成功生产的瓶颈。我们可以通过使用我们最近开发的尖端计算酶设计方法来解决这一挑战，为大肠杆菌创建一条全新的W辅因子生物合成途径。在这个项目中开发的W-BioCat菌株将能够从遗传数据库中表达新的W-酶，并促进新的工程W-酶的生产。这些新的W-酶的催化潜力将被建立并在新的工艺中实施。在生物催化二氧化碳还原为有价值的化学品和芳香族化合物的Birch还原方面，将探索令人振奋的新反应范围，同时还将探索已经确立的和广泛适用的羧酸还原。W-BioCat将是使W-酶可用于工业的突破。作为概念验证，将创建一种氢驱动的过程，将植物来源的油酸转化为润肤剂油酸酯油酸酯。油酸油酯用于数以百万计的人日常使用的许多化妆品中。这一过程将在可扩展的工业相关加氢反应器中以多克产量进行演示，并进行市场研究，以解决商业化的途径。

Europe needs a sustainable chemical industry which will only be realized by new breakthrough technologies. Industrial biotechnology is established in chemical manufacturing, offering more efficient, more specific, safer and less energy demanding production, but is held back by the limited number of enzyme classes in industrial use. This project opens up an important new enzyme class of tungstencontaining enzymes (W-enzymes) which catalyse amazing chemical reactions involving challenging low redox potential reduction reactions, but are currently impossible to obtain economically and on scale to match industrial needs. We need to produce W-enzymes using an industrial workhorse micro-organism such as E. coli. Yet, we discovered that W-cofactor biosynthesisisthe bottleneck preventing successful production of W-enzymes in E. coli. We can solve this challenge by using cutting-edge computational enzyme design approaches we recently developed, to create a completely new W-cofactor biosynthesis pathway for E. coli. The W-BioCat strains developed in this project will enable expression of new W-enzymes from genetic databases, and facilitate production of new engineered W-enzymes. The catalytic potential of these new W-enzymes will be established and implemented in new processes. Exciting new reaction scope in biocatalytic CO2 reduction to valuable chemicals and Birch reduction of aromatic compounds will be explored, alongside the already-established and broadly applicable carboxylic acid reductions. W-BioCat will be the breakthrough to make W-enzymes accessible for industry. As a proof of concept, a hydrogen-driven processto convert plant-derived oleic acid to the emollient ester oleyl oleate will be created. Oleyl oleate is used in many cosmetic products used daily by millions of people. This process will be demonstrated in multi-gram yield in scalable, industrially-relevant hydrogenation reactors, together with market research to address a pathway to commercialisation.