Enhancing the reactivity of a P450 decarboxylase with ionic liquids†

IF 9.3 1区化学 Q1 CHEMISTRY, MULTIDISCIPLINARY

Green Chemistry Pub Date : 2024-11-22 DOI:10.1039/d4gc05292g

Jake H. Nicholson , Mayara Chagas de Avila , Ricardo Rodrigues de Melo , Leticia Maria Zanphorlin , Alex P. S. Brogan

{"title":"Enhancing the reactivity of a P450 decarboxylase with ionic liquids†","authors":"Jake H. Nicholson , Mayara Chagas de Avila , Ricardo Rodrigues de Melo , Leticia Maria Zanphorlin , Alex P. S. Brogan","doi":"10.1039/d4gc05292g","DOIUrl":null,"url":null,"abstract":"<div><div>The cytochrome P450 family of enzymes have been shown to be powerful biocatalysts for a wide range of selective transformations. However, the industrial uptake of P450 enzymes has been low due to issues with enzyme stability and the requirement for exogenous cofactors to drive the reaction. Herein we describe a facile and scalable method for the stabilisation and solubilisation of a P450 decarboxylase enzyme in ionic liquids. The utilisation of ionic liquids allowed for solubilisation of the relatively water-insoluble fatty acid substrate of the enzyme and the modified enzyme was found to be significantly more thermally stable in ionic liquids relative to the enzyme in aqueous media. The shift to non-aqueous solvent allowed for the enzyme to operate in the <em>V</em><sub>max</sub> region, which when coupled with the improved thermal stability, ultimately resulted in a 1000-fold increase in the process intensity of fatty acid decarboxylation. A novel photochemical method for driving the reaction was also discovered which removed the requirement for exogenous H<sub>2</sub>O<sub>2</sub> to be added to the reaction. These results highlight the potential of this strategy as it facilitates a holistic process of biocatalysis engineering where by solvent consideration and increased thermal stability significantly broadens the capability of the enzyme, crucial for the wider realization of industrial biocatalysis.</div></div>","PeriodicalId":78,"journal":{"name":"Green Chemistry","volume":"27 2","pages":"Pages 517-526"},"PeriodicalIF":9.3000,"publicationDate":"2024-11-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.rsc.org/en/content/articlepdf/2025/gc/d4gc05292g?page=search","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Green Chemistry","FirstCategoryId":"92","ListUrlMain":"https://www.sciencedirect.com/org/science/article/pii/S1463926224009397","RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"CHEMISTRY, MULTIDISCIPLINARY","Score":null,"Total":0}

引用次数: 0

Abstract

The cytochrome P450 family of enzymes have been shown to be powerful biocatalysts for a wide range of selective transformations. However, the industrial uptake of P450 enzymes has been low due to issues with enzyme stability and the requirement for exogenous cofactors to drive the reaction. Herein we describe a facile and scalable method for the stabilisation and solubilisation of a P450 decarboxylase enzyme in ionic liquids. The utilisation of ionic liquids allowed for solubilisation of the relatively water-insoluble fatty acid substrate of the enzyme and the modified enzyme was found to be significantly more thermally stable in ionic liquids relative to the enzyme in aqueous media. The shift to non-aqueous solvent allowed for the enzyme to operate in the V_max region, which when coupled with the improved thermal stability, ultimately resulted in a 1000-fold increase in the process intensity of fatty acid decarboxylation. A novel photochemical method for driving the reaction was also discovered which removed the requirement for exogenous H₂O₂ to be added to the reaction. These results highlight the potential of this strategy as it facilitates a holistic process of biocatalysis engineering where by solvent consideration and increased thermal stability significantly broadens the capability of the enzyme, crucial for the wider realization of industrial biocatalysis.

Abstract Image

查看原文本刊更多论文

提高P450脱羧酶与离子液体的反应活性

细胞色素P450酶家族已被证明是广泛选择性转化的强大生物催化剂。然而，由于酶稳定性和需要外源辅助因子来驱动反应的问题，P450酶的工业吸收一直很低。在这里，我们描述了一种简便和可扩展的方法来稳定和溶解离子液体中的P450脱羧酶。离子液体的使用允许对酶的相对不溶于水的脂肪酸底物进行增溶，并且发现修饰后的酶在离子液体中相对于在水介质中的酶具有更大的热稳定性。向非水溶剂的转变允许酶在Vmax区域工作，这与改进的热稳定性相结合，最终导致脂肪酸脱羧过程强度增加1000倍。还发现了一种新的光化学方法来驱动该反应，该方法不需要在反应中添加外源H2O2。这些结果突出了该策略的潜力，因为它促进了生物催化工程的整体过程，其中通过溶剂考虑和增加的热稳定性显着拓宽了酶的能力，这对于更广泛地实现工业生物催化至关重要。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Green Chemistry 化学-化学综合

CiteScore

16.10

自引率

7.10%

发文量

677

审稿时长

1.4 months

期刊介绍： Green Chemistry is a journal that provides a unique forum for the publication of innovative research on the development of alternative green and sustainable technologies. The scope of Green Chemistry is based on the definition proposed by Anastas and Warner (Green Chemistry: Theory and Practice, P T Anastas and J C Warner, Oxford University Press, Oxford, 1998), which defines green chemistry as the utilisation of a set of principles that reduces or eliminates the use or generation of hazardous substances in the design, manufacture and application of chemical products. Green Chemistry aims to reduce the environmental impact of the chemical enterprise by developing a technology base that is inherently non-toxic to living things and the environment. The journal welcomes submissions on all aspects of research relating to this endeavor and publishes original and significant cutting-edge research that is likely to be of wide general appeal. For a work to be published, it must present a significant advance in green chemistry, including a comparison with existing methods and a demonstration of advantages over those methods.