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香菇磷酸葡萄糖异构酶的克隆与特性分析。

Cloning and characterization of phosphoglucose isomerase in Lentinula edodes.

机构信息

Key Laboratory of Microbiological Engineering of Agricultural Environment, Ministry of Agriculture, Department of Microbiology, College of Life Sciences, Nanjing Agricultural University, Nanjing, Jiangsu, P.R. China.

出版信息

J Basic Microbiol. 2022 Jun;62(6):740-749. doi: 10.1002/jobm.202100598. Epub 2022 Feb 23.

Abstract

Phosphoglucose isomerase (PGI) is a key enzyme that participates in polysaccharide synthesis, which is responsible for the interconversion of glucose-6-phosphate (G-6-P) and fructose-6-phosphate (F-6-P), but there is little research focusing on its role in fungi, especially in higher basidiomycetes. The pgi gene was cloned from Lentinula edodes and named lepgi. Then, the lepgi-silenced strains were constructed by RNA interference. In this study, we found that lepgi-silenced strains had significantly less biomass than the wild-type (WT) strain. Furthermore, the extracellular polysaccharide (EPS) and intracellular polysaccharide (IPS) levels increased 1.5- to 3-fold and 1.5-fold, respectively, in lepgi-silenced strains. Moreover, the cell wall integrity in the silenced strains was also altered, which might be due to changes in the compounds and structure of the cell wall. The results showed that compared to WT, silencing lepgi led to a significant decrease of approximately 40% in the β-1,3-glucan content, and there was a significant increase of 2-3-fold in the chitin content. These findings provide support for studying the biological functions of lepgi in L. edodes.

摘要

磷酸葡萄糖异构酶(PGI)是一种参与多糖合成的关键酶,负责葡萄糖-6-磷酸(G-6-P)和果糖-6-磷酸(F-6-P)的相互转化,但关于其在真菌中的作用,特别是在高等担子菌中的作用的研究甚少。本研究从香菇中克隆了 pgi 基因,并将其命名为 lepgi。然后通过 RNA 干扰构建 lepgi 沉默菌株。在这项研究中,我们发现 lepgi 沉默菌株的生物量明显低于野生型(WT)菌株。此外,lepgi 沉默菌株的胞外多糖(EPS)和胞内多糖(IPS)水平分别增加了 1.5-3 倍和 1.5 倍。此外,沉默菌株的细胞壁完整性也发生了改变,这可能是由于细胞壁化合物和结构的变化。结果表明,与 WT 相比,沉默 lepgi 导致β-1,3-葡聚糖含量显著下降约 40%,几丁质含量显著增加 2-3 倍。这些发现为研究 lepgi 在香菇中的生物学功能提供了支持。

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