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在葡萄糖、稻草、锯末、小麦籽粒和混合基质上生长的生理特性及分泌蛋白质组比较分析

Physiological Characteristics and Comparative Secretome Analysis of Grown on Glucose, Rice Straw, Sawdust, Wheat Grain, and MIX Substrates.

作者信息

Cai YingLi, Ma XiaoLong, Zhang QianQian, Yu FuQiang, Zhao Qi, Huang Wei, Song JiaXin, Liu Wei

机构信息

Institute of Vegetable, Wuhan Academy of Agricultural Sciences, Wuhan, China.

Institute of Applied Mycology, Huazhong Agricultural University, Wuhan, China.

出版信息

Front Microbiol. 2021 May 25;12:636344. doi: 10.3389/fmicb.2021.636344. eCollection 2021.

Abstract

Morels ( sp.) are economically important edible macro-fungi, which can grow on various synthetic or semi-synthetic media. However, the complex nutritional metabolism and requirements of these fungi remain ill-defined. This study, based on the plant biomass commonly used in the artificial cultivation of morels, assessed and compared the growth characteristics and extracellular enzymes of cultivated on glucose, rice straw, sawdust, wheat grain, and a mixture of equal proportions of the three latter plant substrates (MIX). could grow on all five tested media but displayed significant variations in mycelial growth rate, biomass, and sclerotium yield on the different media. The most suitable medium for was wheat and wheat-containing medium, followed by glucose, while rice straw and sawdust were the least suitable. A total of 268 secretory proteins were identified by liquid chromatography coupled with tandem mass spectrometry detection. Functional classification and label-free comparative analysis of these proteins revealed that carbohydrate-active enzyme (CAZYme) proteins were the predominant component of the secretome of , followed by protease, peptidase, and other proteins. The abundances of CAZYme proteins differed among the tested media, ranging from 64% on glucose to 88% on rice straw. The CAZYme classes of glycoside hydrolases and carbohydrate-binding module were enriched in the five secretomes. Furthermore, the enzyme activities of CMCase, lignase, amylase, xylase, pNPCase, and pNPGase were detected during the continuous culture of in MIX medium, and the relative expression of the corresponding genes were detected by quantitative real-time PCR. The combined data of growth potential, secretome, extracellular enzyme activity, and gene expression on different substrates inferred that was weak in lignocellulose degradation but a good starch decomposer. Specifically, in terms of the degradation of cellulose, the ability to degrade cellulose into oligosaccharides was weaker compared with further degradation into monosaccharides, and this might be the speed-limiting step of cellulose utilization in . In addition, had a strong ability to decompose various hemicellulose glycosidic bonds, especially α- and β-galactosidase. Only a very few lignin-degradation-related proteins were detected, and these were in low abundance, consistent with the presence of weak lignin degradation ability. Furthermore, the presence of lipase and chitinase implied that was capable of decomposition of its own mycelia . The study provides key data that facilitates a further understanding of the complex nutritional metabolism of .

摘要

羊肚菌(羊肚菌属)是具有重要经济价值的可食用大型真菌,能够在各种合成或半合成培养基上生长。然而,这些真菌复杂的营养代谢和需求仍不明确。本研究基于羊肚菌人工栽培中常用的植物生物质,评估并比较了在葡萄糖、稻草、锯末、小麦粒以及后三种植物基质等比例混合物(MIX)上培养的羊肚菌的生长特性和胞外酶。羊肚菌能够在所有五种测试培养基上生长,但在不同培养基上其菌丝生长速率、生物量和菌核产量存在显著差异。最适合羊肚菌生长的培养基是小麦及含小麦的培养基,其次是葡萄糖,而稻草和锯末最不适合。通过液相色谱联用串联质谱检测共鉴定出268种分泌蛋白。对这些蛋白的功能分类和无标记比较分析表明,碳水化合物活性酶(CAZYme)蛋白是羊肚菌分泌组的主要成分,其次是蛋白酶、肽酶和其他蛋白。不同测试培养基中CAZYme蛋白的丰度有所不同,从葡萄糖上的64%到稻草上的88%不等。糖苷水解酶和碳水化合物结合模块的CAZYme类别在五个分泌组中富集。此外,在MIX培养基中对羊肚菌进行连续培养期间检测了羧甲基纤维素酶(CMCase)、木质素酶、淀粉酶、木聚糖酶、对硝基苯磷酸二酯酶(pNPCase)和对硝基苯β - 葡萄糖苷酶(pNPGase)的酶活性,并通过实时定量PCR检测了相应基因的相对表达。不同底物上生长潜力、分泌组、胞外酶活性和基因表达的综合数据推断,羊肚菌在木质纤维素降解方面较弱,但淀粉分解能力良好。具体而言,在纤维素降解方面,将纤维素降解为寡糖的能力比进一步降解为单糖的能力弱,这可能是羊肚菌利用纤维素的限速步骤。此外,羊肚菌具有很强的分解各种半纤维素糖苷键的能力,尤其是α - 和β - 半乳糖苷酶。仅检测到极少数与木质素降解相关的蛋白,且这些蛋白丰度较低,这与羊肚菌木质素降解能力较弱一致。此外,脂肪酶和几丁质酶的存在表明羊肚菌能够分解自身的菌丝体。该研究提供了关键数据,有助于进一步了解羊肚菌复杂的营养代谢。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7db0/8185036/89afda0f14a4/fmicb-12-636344-g001.jpg

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