Exploring the biotechnological potential of the UbiD (de)carboxylase family: novel biochemistry for renewable commodity chemicals

探索 UbiD（脱）羧酶家族的生物技术潜力：可再生商品化学品的新型生物化学

• 批准号: BB/P000622/1
• 负责人: David Leys
• 金额: $ 74.61万
• 依托单位: University of Manchester
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2017
• 资助国家: 英国
• 起止时间: 2017 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FP000622%2F1
• 关键词: Exploring; biotechnological; potential; UbiD; de

The projected depletion of fossil oil resources, and the threat of global warming as a direct consequence of their continued use, has led to the challenge of providing alternative and renewable routes to oil derived fuels and commodity chemicals. However, any such process faces severe demands in terms of yield and costs before it can be considered industrially relevant. In addition, the production of oil derived compounds (ie hydrocarbons) or their functionalization does not belong to what can be considered the "standard" biochemical repertoire. Hence, comparably few biocatalysts, pathways or organisms have been reported to might support such processes. Therefore, while the challenge is clearly defined, the route to a suitable solution if far from clear, and there is an unmet industrial need for new biocatalysts that can produce commodity chemicals such as butadiene (precursor to synthetic rubber) and aromatic dicarboxylic acids (precursors to PET and other polymers). We have recently been able to provide some detailed insights into the mechanism of the UbiD family of enzymes. These act as reversible decarboxylates (ie adding or removing a CO2 group to/from an organic molecule). We and others have shown they are able to interconvert unsaturated hydrocarbons (ie containing a double bond) with the corresponding carboxylic acids (ie version containing the additional CO2 group). A wide range of substrates has been reported, and the reaction catalysed appears readily reversible depending on [CO2] levels. Our recent work on these enzyme has established the achieve this unusual chemistry by making use of a previously unknown enzyme cofactor. In this project, we seek to complete our understanding of how this cofactor is made, and also how it is inactivated, to guide future application.Building on our fundamental understanding, we seek to derive novel and green production routes to commodity chemicals using renewable feedstocks as UbiD substrates. More specifically, in this grant we will seek to establish to what extend UbiD enzyme can be evolved to produce compounds such as butadiene and aromatic dicarboxylic acids. We will make use of the renewable feedstocks muconic and benzoic acid as UbiD substrates respectively. Our results will provide proof of concept and outline the scope for future biotechnological application.

化石石油资源的预计枯竭，以及继续使用化石石油资源直接造成的全球变暖威胁，导致了为石油衍生燃料和商品化学品提供替代和可再生途径的挑战。然而，在被认为具有工业意义之前，任何这样的工艺在产量和成本方面都面临着严峻的要求。此外，石油衍生化合物(即碳氢化合物)的生产或其官能化不属于可被认为是“标准”的生化谱系。因此，据报道可能支持这一过程的生物催化剂、途径或生物体相对较少。因此，虽然挑战已明确界定，但获得适当解决方案的途径远未明确，而且工业上对能够生产诸如丁二烯(合成橡胶的前体)和芳香族二元酸(聚酯和其他聚合物的前体)等商品化学品的新型生物催化剂的需求尚未得到满足。我们最近能够提供一些关于UbiD酶家族的机制的详细见解。它们起到可逆脱羧酸盐的作用(即在有机分子中添加或移除二氧化碳基团)。我们和其他人已经证明，他们能够将不饱和碳氢化合物(即包含双键)与相应的羧酸(即包含额外二氧化碳基团的版本)相互转化。已有大量的底物被报道，根据[CO2]水平的不同，催化的反应似乎很容易逆转。我们最近对这些酶的研究已经确立了通过利用一种以前未知的酶辅助因子来实现这种不同寻常的化学作用。在这个项目中，我们试图完成我们对这种辅因子是如何产生的，以及它是如何被灭活的理解，以指导未来的应用。在我们的基本理解的基础上，我们寻求利用可再生原料作为UBD底物来获得用于商品化学品的新颖和绿色的生产路线。更具体地说，在这项资助中，我们将寻求建立UbiD酶可以进化到什么程度来产生丁二烯和芳香族二元酸等化合物。我们将分别使用可再生原料Muconic和苯甲酸作为UbiD底物。我们的结果将提供概念证明，并勾勒出未来生物技术应用的范围。

项目成果

Enzymatic N-Allylation of Primary and Secondary Amines Using Renewable Cinnamic Acids Enabled by Bacterial Reductive Aminases.

• DOI: 10.1021/acssuschemeng.2c01180
• 发表时间: 2022-05-23
• 期刊: ACS SUSTAINABLE CHEMISTRY & ENGINEERING
• 影响因子: 8.4
• 作者: Aleku, Godwin A.;Titchiner, Gabriel R.;Roberts, George W.;Derrington, Sasha R.;Marshall, James R.;Hollfelder, Florian;Turner, Nicholas J.;Leys, David
• 通讯作者: Leys, David

The role of conserved residues in Fdc decarboxylase in prenylated flavin mononucleotide oxidative maturation, cofactor isomerization, and catalysis.

• DOI: 10.1074/jbc.ra117.000881
• 发表时间: 2018-02-16
• 期刊: The Journal of biological chemistry
• 作者: Bailey SS;Payne KAP;Fisher K;Marshall SA;Cliff MJ;Spiess R;Parker DA;Rigby SEJ;Leys D
• 通讯作者: Leys D

Structural and catalytic properties of the peroxygenase P450 enzyme CYP152K6 from Bacillus methanolicus.

• DOI: 10.1016/j.jinorgbio.2018.08.002
• 发表时间: 2018-11
• 期刊: Journal of inorganic biochemistry
• 影响因子: 3.9
• 作者: Girvan HM;Poddar H;McLean KJ;Nelson DR;Hollywood KA;Levy CW;Leys D;Munro AW
• 通讯作者: Munro AW