Tungsten Biocatalysis - Heavy Metal Enzymes for Sustainable Industrial Biocatalysis

欧洲需要一个可持续的化学工业，而这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业，提供更有效、更具体、更安全和能源需求更少的生产，但由于工业使用的酶类数量有限而受到阻碍。该项目开辟了一种重要的新型酶类——含钨酶（w -酶），它可以催化令人惊叹的化学反应，包括具有挑战性的低氧化还原电位还原反应，但目前还无法在经济上和规模上获得以满足工业需求。我们需要使用像大肠杆菌这样的工业主力微生物来生产w酶。然而，我们发现w -辅因子的生物合成是阻碍大肠杆菌成功生产w -酶的瓶颈。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个挑战，为大肠杆菌创造一个全新的w -辅因子生物合成途径。本项目开发的W-BioCat菌株将能够从基因数据库中表达新的w -酶，并促进新的w -酶工程的生产。这些新w酶的催化潜力将在新的工艺中得到建立和实现。在生物催化CO2还原到有价值的化学品和桦木还原芳香族化合物的令人兴奋的新反应范围将被探索，以及已经建立和广泛适用的羧酸还原。W-BioCat将成为w酶进入工业应用的突破口。作为概念的验证，将创建一个氢驱动的过程，将植物衍生的油酸转化为润肤酯油酸酯。油酸酯被用于数百万人日常使用的许多化妆品中。这一过程将在可扩展的、与工业相关的加氢反应器中以多克产量进行演示，同时进行市场研究，以解决商业化途径。

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.