黑曲霉水杨酸羟化酶的发现与功能分析。
Discovery and Functional Analysis of a Salicylic Acid Hydroxylase from Aspergillus niger.
机构信息
Fungal Physiology, Westerdijk Fungal Biodiversity Institute & Fungal Molecular Physiology, Utrecht University, Utrecht, The Netherlands.
Department of Microbiology, University of Helsinki, Helsinki, Finland.
出版信息
Appl Environ Microbiol. 2021 Feb 26;87(6). doi: 10.1128/AEM.02701-20.
Salicylic acid plays an important role in the plant immune response, and its degradation is therefore important for plant-pathogenic fungi. However, many nonpathogenic microorganisms can also degrade salicylic acid. In the filamentous fungus , two salicylic acid metabolic pathways have been suggested. The first pathway converts salicylic acid to catechol by a salicylate hydroxylase (ShyA). In the second pathway, salicylic acid is 3-hydroxylated to 2,3-dihydroxybenzoic acid, followed by decarboxylation to catechol by 2,3-dihydroxybenzoate decarboxylase (DhbA). cleaves the aromatic ring of catechol catalyzed by catechol 1,2-dioxygenase (CrcA) to form ,-muconic acid. However, the identification and role of the genes and characterization of the enzymes involved in these pathways are lacking. In this study, we used transcriptome data of grown on salicylic acid to identify genes ( and ) involved in salicylic acid metabolism. Heterologous production in followed by biochemical characterization confirmed the function of ShyA and CrcA. The combination of ShyA and CrcA demonstrated that -muconic acid can be produced from salicylic acid. In addition, the roles of , , and were studied by creating deletion mutants which revealed the role of these genes in the fungal metabolism of salicylic acid. Nonrenewable petroleum sources are being depleted, and therefore, alternative sources are needed. Plant biomass is one of the most abundant renewable sources on Earth and is efficiently degraded by fungi. In order to utilize plant biomass efficiently, knowledge about the fungal metabolic pathways and the genes and enzymes involved is essential to create efficient strategies for producing valuable compounds such as ,-muconic acid. ,-Muconic acid is an important platform chemical that is used to synthesize nylon, polyethylene terephthalate (PET), polyurethane, resins, and lubricants. Currently, ,-muconic acid is mainly produced through chemical synthesis from petroleum-based chemicals. Here, we show that two enzymes from fungi can be used to produce ,-muconic acid from salicylic acid and contributes in creating alternative methods for the production of platform chemicals.
水杨酸在植物免疫反应中起着重要作用，因此其降解对植物病原真菌很重要。然而，许多非致病微生物也可以降解水杨酸。在丝状真菌中，已经提出了两种水杨酸代谢途径。第一条途径通过水杨酸羟化酶（ShyA）将水杨酸转化为儿茶酚。在第二条途径中，水杨酸 3-羟基化为 2,3-二羟基苯甲酸，然后 2,3-二羟基苯甲酸脱羧酶（DhbA）将其脱羧为儿茶酚。儿茶酚 1,2-双加氧酶（CrcA）催化断裂儿茶酚的芳环，形成 -粘康酸。然而，这些途径中涉及的基因的鉴定和作用以及酶的表征尚不清楚。在这项研究中，我们使用在水杨酸上生长的转录组数据来鉴定参与水杨酸代谢的基因（和）。在 中的异源生产随后的生化特征证实了 ShyA 和 CrcA 的功能。ShyA 和 CrcA 的组合表明可以从水杨酸生产 -粘康酸。此外，通过创建 缺失突变体研究了 、 和 的 作用，这揭示了这些基因在真菌水杨酸代谢中的作用。不可再生的石油资源正在枯竭，因此需要替代资源。植物生物质是地球上最丰富的可再生资源之一，被真菌有效地降解。为了有效地利用植物生物质，了解真菌代谢途径以及相关的基因和酶对于创造生产有价值化合物（如 -粘康酸）的有效策略至关重要。-粘康酸是一种重要的平台化学品，用于合成尼龙、聚对苯二甲酸乙二醇酯（PET）、聚氨酯、树脂和润滑剂。目前，-粘康酸主要通过从石油化工产品的化学合成生产。在这里，我们展示了真菌中的两种酶可以用于从水杨酸生产 -粘康酸，并有助于创造生产平台化学品的替代方法。
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