Feruloyl-CoA synthetases and feruloyl-CoA hydratase/lyases: Expression, biochemical characterisation, and generation of vanillin from ferulic acid and lignocellulosic hydrolysates.

4区生物学 Q3 Biochemistry, Genetics and Molecular Biology

Methods in enzymology Pub Date : 2025-01-01 Epub Date: 2025-02-16 DOI:10.1016/bs.mie.2025.01.046

Victoria Sodré, Fabio Marcio Squina

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引用次数: 0

Abstract

Vanillin (4-hydroxy-3-methoxybenzaldehyde) is a valuable aroma and flavour compound with diverse applications in the food, fragrance, and pharmaceutical industries. Currently, the majority of vanillin produced globally is chemically synthesised from fossil-derived resources, but biocatalytic production from plant biomass offers a sustainable alternative. Alkaline pretreatment of grass-derived biomass releases ferulic acid, which can be converted into vanillin through a two-step biotransformation catalysed by feruloyl-CoA synthetases (FCSs) and feruloyl-CoA hydratase/lyases (FCHLs). This article presents detailed methodologies for the expression, purification, and biochemical characterisation of FCSs and FCHLs sourced from a lignin-degrading microbial consortium. Additionally, it describes protocols for preparing alkaline pretreatment hydrolysates from sugarcane bagasse and implementing the coupled FCS/FCHL reaction for vanillin synthesis. Analytical techniques for monitoring substrates and products are also discussed. These methods aim to support researchers in advancing the biocatalytic production of vanillin from renewable plant biomass.

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阿魏酸辅酶a合成酶和阿魏酸辅酶a水合酶/裂解酶：从阿魏酸和木质纤维素水解物中表达、生化特性和生成香兰素。

香兰素（4-羟基-3-甲氧基苯甲醛）是一种有价值的芳香和风味化合物，在食品、香料和制药工业中有着广泛的应用。目前，全球生产的大多数香兰素都是由化石来源的资源化学合成的，但植物生物质的生物催化生产提供了一种可持续的替代方案。草源生物质的碱性预处理释放阿魏酸，阿魏酸可以通过阿魏酰辅酶a合成酶（FCSs）和阿魏酰辅酶a水合酶/裂解酶（FCHLs）催化的两步生物转化转化为香兰素。本文介绍了来自木质素降解微生物群落的FCSs和FCHLs的表达、纯化和生化表征的详细方法。此外，它还描述了从蔗渣中制备碱性预处理水解物和实现FCS/FCHL耦合反应合成香兰素的方案。还讨论了监测基材和产品的分析技术。这些方法旨在支持研究人员推进可再生植物生物质生物催化生产香兰素。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Methods in enzymology 生物-生化研究方法

CiteScore

2.90

自引率

0.00%

发文量

308

审稿时长

3-6 weeks

期刊介绍： The critically acclaimed laboratory standard for almost 50 years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Each volume is eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. Now with over 500 volumes the series contains much material still relevant today and is truly an essential publication for researchers in all fields of life sciences, including microbiology, biochemistry, cancer research and genetics-just to name a few. Five of the 2013 Nobel Laureates have edited or contributed to volumes of MIE.