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调查定殖于大麦籽粒中的土著真菌群落:分泌组和木聚糖酶潜力。

Investigation of the indigenous fungal community populating barley grains: Secretomes and xylanolytic potential.

机构信息

Enzyme and Protein Chemistry, Department of Biotechnology and Biomedicine, Technical University of Denmark, Elektrovej, Building 375, DK-2800 Kgs. Lyngby, Denmark.

Fungal Chemodiversity, Department of Biotechnology and Biomedicine, Technical University of Denmark, Søltofts Plads, Building 221, DK -2800 Kgs. Lyngby, Denmark.

出版信息

J Proteomics. 2017 Oct 3;169:153-164. doi: 10.1016/j.jprot.2017.03.009. Epub 2017 Mar 18.

Abstract

UNLABELLED

The indigenous fungal species populating cereal grains produce numerous plant cell wall-degrading enzymes including xylanases, which could play important role in plant-pathogen interactions and in adaptation of the fungi to varying carbon sources. To gain more insight into the grain surface-associated enzyme activity, members of the populating fungal community were isolated, and their secretomes and xylanolytic activities assessed. Twenty-seven different fungal species were isolated from grains of six barley cultivars over different harvest years and growing sites. The isolated fungi were grown on medium containing barley flour or wheat arabinoxylan as sole carbon source. Their secretomes and xylanase activities were analyzed using SDS-PAGE and enzyme assays and were found to vary according to species and carbon source. Secretomes were dominated by cell wall degrading enzymes with xylanases and xylanolytic enzymes being the most abundant. A 2-DE-based secretome analysis of Aspergillus niger and the less-studied pathogenic fungus Fusarium poae grown on barley flour and wheat arabinoxylan resulted in identification of 82 A. niger and 31 F. poae proteins many of which were hydrolytic enzymes, including xylanases.

BIOLOGICAL SIGNIFICANCE

The microorganisms that inhabit the surface of cereal grains are specialized in production of enzymes such as xylanases, which depolymerize plant cell walls. Integration of gel-based proteomics approach with activity assays is a powerful tool for analysis and characterization of fungal secretomes and xylanolytic activities which can lead to identification of new enzymes with interesting properties, as well as provide insight into plant-fungal interactions, fungal pathogenicity and adaptation. Understanding the fungal response to host niche is of importance to uncover novel targets for potential symbionts, anti-fungal agents and biotechnical applications.

摘要

未加标签

定殖于谷物中的本土真菌物种产生许多植物细胞壁降解酶,包括木聚糖酶,这些酶在植物-病原体相互作用以及真菌适应不同碳源方面可能发挥重要作用。为了更深入地了解谷物表面相关的酶活性,分离了定殖真菌群落的成员,并评估了它们的分泌组和木聚糖酶活性。从六个不同收获年份和种植地点的大麦品种的谷物中分离出了 27 种不同的真菌。将分离出的真菌在含有大麦粉或小麦阿拉伯木聚糖作为唯一碳源的培养基上培养。使用 SDS-PAGE 和酶测定法分析了它们的分泌组和木聚糖酶活性,发现这些活性因物种和碳源而异。分泌组主要由细胞壁降解酶组成,其中木聚糖酶和木聚糖降解酶最为丰富。基于 2-DE 的黑曲霉和研究较少的致病真菌禾谷镰刀菌在大麦粉和小麦阿拉伯木聚糖上生长的分泌组分析,鉴定出了 82 种黑曲霉和 31 种禾谷镰刀菌蛋白,其中许多是水解酶,包括木聚糖酶。

生物学意义

定殖于谷物表面的微生物专门生产木聚糖酶等酶,这些酶可分解植物细胞壁。将凝胶基蛋白质组学方法与活性测定相结合,是分析和表征真菌分泌组和木聚糖酶活性的有力工具,可导致发现具有有趣特性的新酶,并深入了解植物-真菌相互作用、真菌致病性和适应性。了解真菌对宿主小生境的反应对于揭示潜在共生体、抗真菌剂和生物技术应用的新靶标具有重要意义。

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