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通过磷酸葡萄糖变位酶的靶向过表达增强瓦尼桑黄胞外多糖的合成及其对多糖生物功能特性的影响。

Augmentation of exopolysaccharide synthesis and its influence on biofunctional properties of polysaccharide in Sanghuangporus vaninii via targeted overexpression of phosphoglucomutase.

作者信息

Li Zihao, Xu Congtao, Pan Jinlong, Zhou Yi, Li Haikang, Li Weihang, Zou Yajie

机构信息

State Key Laboratory of Efficient Utilization of Arable Land in China, Institute of Agricultural Resources and Regional Planning, Chinese Academy of Agricultural Sciences, Beijing 100081, China; School of Ecology and Nature Conservation, Beijing Forestry University, Beijing 100083, China.

State Key Laboratory of Efficient Utilization of Arable Land in China, Institute of Agricultural Resources and Regional Planning, Chinese Academy of Agricultural Sciences, Beijing 100081, China.

出版信息

Int J Biol Macromol. 2025 May;306(Pt 2):141182. doi: 10.1016/j.ijbiomac.2025.141182. Epub 2025 Feb 16.

Abstract

Phosphoglucomutase (PGM) is considered an essential catalyst in polysaccharide biosynthesis, plays a pivotal role in the early stage of carbon metabolism and the synthesis of nucleotide sugars. Polysaccharides are important active components of Sanghuangporus vaninii. Based on the transcriptomic data, the successful cloning and re-annotation of the putative 1353 - bp PGM took place, which encodes a protein of 49.45 kDa. In order to elucidate the function of this gene, overexpression and interference transformation systems were constructed in S. vaninii. The results revealed that the PGM is primarily responsible for regulating the interconversion between glucose-1-phosphate and glucose-6-phosphate. An up - regulation of PGM expression resulted in a significant increase in extracellular polysaccharide content, quantified at 7.72 % and 15.15 %, whereas a down - regulation of PGM expression resulted in a significant increase in intracellular polysaccharide content (33.68 % and 38.24 %). The antioxidant capacity of intracellular polysaccharide in strain, including hydroxyl radical scavenging, superoxide anion radical scavenging, 2-diphenyl-1-picrylhydrazyl (DPPH) scavenging, ABTS radical scavenging and FRAP scavenging was significantly increased by RNA interference with the expression of PGM, while the overexpression of PGM was found to enhance the antioxidant capacity of extracellular polysaccharide to a certain degrees. Furthermore, the modulation of PGM expression had a significant impact on polysaccharide metabolic pathways, resulting in alterations in monosaccharide composition and impacting the content of cell wall components, and the microstructure of polysaccharides. These findings elucidate the polysaccharide biosynthetic pathway in S. vaninii, highlighting the PGM as a prime target for the propagation of high-yield polysaccharide-producing strains. This study stands as a basic reference for amplifying polysaccharide production via strategic metabolic regulation.

摘要

磷酸葡萄糖变位酶(PGM)被认为是多糖生物合成中的一种必需催化剂,在碳代谢的早期阶段和核苷酸糖的合成中起关键作用。多糖是瓦尼桑黄的重要活性成分。基于转录组数据,成功克隆并重新注释了推定的1353 bp的PGM,其编码一种49.45 kDa的蛋白质。为了阐明该基因的功能,在瓦尼桑黄中构建了过表达和干扰转化系统。结果表明,PGM主要负责调节1-磷酸葡萄糖和6-磷酸葡萄糖之间的相互转化。PGM表达上调导致细胞外多糖含量显著增加,分别为7.72%和15.15%,而PGM表达下调导致细胞内多糖含量显著增加(33.68%和38.24%)。通过RNA干扰PGM的表达,菌株中细胞内多糖的抗氧化能力显著提高,包括羟基自由基清除、超氧阴离子自由基清除、2,2-二苯基-1-苦基肼(DPPH)清除、ABTS自由基清除和铁还原抗氧化能力(FRAP)清除,而PGM的过表达在一定程度上增强了细胞外多糖的抗氧化能力。此外,PGM表达的调节对多糖代谢途径有显著影响,导致单糖组成改变,影响细胞壁成分的含量以及多糖的微观结构。这些发现阐明了瓦尼桑黄中的多糖生物合成途径,突出了PGM作为高产多糖生产菌株繁殖的主要靶点。本研究为通过战略代谢调控扩大多糖产量提供了基础参考。

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