X-Genix: Translating Halogenases for Sustainable Synthesis

1a.i.概念验证的概念简介：90%以上的药物的制造包括卤化步骤，将卤素添加到碳中，形成所谓的C-X键。对200种最畅销药物的分析表明，25%的药物在最终产品中含有这种卤素(X因子)，包括治疗癌症、糖尿病、高胆固醇、胃溃疡、贫血、哮喘、癫痫等疾病的药物。另有67%的人在制造过程中使用C-X键(见图2)。问题：虽然选择性地生成C-X键的能力是必不可少的，但目前实现这一点的化学卤化方法效率低、成本高，并且需要有毒的化学品。它们往往伴随着较差的选择性，从而导致非选择性的卤化和不受欢迎的副产品，在下游净化过程中造成困难、对环境有害的废物和具有成本影响的贵重材料的损失(图1)。现有的芳香族底物卤化方法通常使用高活性试剂，通常产生的产品要么只对最亲核的位置进行卤化，要么产生多种产品的混合物。此外，卤化试剂的生成是一个能源密集型过程；例如，目前仅制药所需的用于生产氯气的能源就会产生400,000吨二氧化碳排放(相当于每年超过87000辆汽车的二氧化碳排放)。在我们发现具有广泛底物范围的新型卤代酶的专利方法之前，还没有适合工业卤化的酶可用。图1：当前化学卤化的主要限制-有毒试剂、非选择性化学、不利的安全和环境影响。机会和突破创新潜力：我们的X-Genix项目提供酶卤化技术，并使用天然工具(定制的酶和盐)选择性地安装C-X键，没有浪费，同时降低成本(图3)。我们的USP是找到黄素依赖卤化酶(FDHS)的专利方法，这是一个获得专利的卤素酶工具箱，以及我们在生产、工程和使用这些可持续酶以实现更绿色、更廉价和更安全的卤化过程的技术诀窍。为什么是现在？在工业中，有一个关键的推动因素，即用酶反应取代化学催化的反应，目标是到2050年，所有工业反应中应该有30%是由酶驱动的。值得注意的是，卤化反应是最重要的转化之一，在工业生物催化组合中仍然缺失。这项提案解决了这一赤字问题。

1a.i. Brief description of the idea to be taken to proof of concept:The manufacture of over 90% of pharmaceuticals includes a halogenation step, addition of a halogen to a carbon, forming a so-called C-X bond. Analysis of 200 top-selling drugs indicated that > 25% of pharmaceuticals contain this halogen (X-factor) in the final product, including drugs for treatment for conditions such as cancer, diabetes, high cholesterol, stomach ulcers, anaemia, asthma, epilepsy, and others. A further 67% use C-X formation in their manufacture (see Fig. 2).The problem: Whilst the ability to selectively make C-X bonds is essential, current chemical halogenation methods to achieve this are inefficient, expensive and require toxic chemicals. They are often accompanied by poor selectivity, which results in non-selective halogenation and undesired by-products, creating difficulties in the downstream purification process, environmentally detrimental waste and loss of precious material with cost implications (Fig.1). Existing methods for halogenating aromatic substrates generally employ highly reactive reagents, which often generate products in which either only the most nucleophilic position is halogenated or mixtures of products are produced.In addition, generation of halogenating reagents is an energy intensive process; for example, energy used for producing chlorine gas currently required for pharmaceutical manufacture alone accounts for >400,000 metric tons of CO2-emission (equating to CO2-emission of over 87000 cars per annum). Enzymes suitable for industrial halogenation had not been available prior to our patented approach to discovery of novel halogenases with broad substrate scope.Figure 1: Major limitations of current chemical halogenations - toxic reagents, non-selective chemistries, adverse safety and environmental impact.The Opportunity and Breakthrough Innovation Potential: Our X-Genix project provides enzymatic halogenation technology and uses natural tools (bespoke enzymes and salt) to selectively install C-X bonds without waste and whilst reducing costs (Fig. 3). Our USP is the patented approach to finding flavin dependent halogenases (FDHs), a patented toolbox of halogenases and our knowhow in producing, engineering and using these sustainable enzymes for greener, cheaper and safer halogenation processes. Why now? There is a critical push in industry toward displacing chemically catalysed reactions with enzymatic reactions, the ambition being that by 2050, 30% of all industrial reactions should be enzyme driven. Notably, halogenation, one of the most important transformations remains missing from the industrial biocatalysis portfolio. This proposal addresses this deficit.