植物乳杆菌YM-4-3菌株胞外多糖合成过程中两种糖基转移酶的分子调控机制

IF 2.5 3区化学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Carbohydrate Research Pub Date : 2025-09-18 DOI:10.1016/j.carres.2025.109676

Zhen Zhang , Bo Zhao , Yonggang Xiong , Xiaoran Li , Chenjian Liu , Junwei Xu , Jinping Yan , En Yang

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引用次数: 0

摘要

外多糖（EPS）是乳酸菌在生长代谢过程中产生的一种高分子量聚合物。由于菌株的遗传多样性和EPS的结构多样性，使其表现出多种生理活性，在食品、医药、农业等领域具有潜在的应用前景。本研究对植物乳杆菌m -4-3菌株EPS生物合成过程中参与低聚糖重复单元合成的两种糖基转移酶（orf1595和cps4I）进行了研究。这些酶是多糖合成的关键。通过敲除和补充orf1595和cps4I基因，结合转录组分析，我们初步确定了它们的功能。结果表明，基因敲除后EPS产量显著下降，而基因互补后EPS产量恢复。转录组测序显示，敲除这些糖基转移酶基因导致EPS合成簇内基因的差异表达，并影响糖代谢途径。此外，EPS抗氧化活性显著改变：清除DPPH自由基和超氧阴离子的能力下降，而清除羟基自由基的能力增强。化学分析显示敲除后EPS蛋白、葡萄糖醛酸和硫酸盐含量发生变化。本研究阐明了orf1595和cps4I在L. plantarum m -4-3合成EPS中的关键作用，为调控EPS合成和促进具有多种生理活性的LAB EPS的发育提供了理论依据。

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Molecular regulation mechanism of two glycosyltransferases in exopolysaccharide biosynthesis of Lactiplantibacillus plantarum YM-4-3 strain

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Molecular regulation mechanism of two glycosyltransferases in exopolysaccharide biosynthesis of Lactiplantibacillus plantarum YM-4-3 strain

Exopolysaccharide (EPS) is a high molecular weight polymer produced by lactic acid bacteria (LAB) during their growth and metabolism. Due to the genetic diversity of strains and the structural diversity of EPS, it exhibits a variety of physiological activities and has potential applications in food, medicine, agriculture, and other fields. This study focuses on two glycosyltransferases (orf1595 and cps4I) involved in the synthesis of oligosaccharide repeat units in the EPS biosynthesis by Lactiplantibacillus plantarum YM-4-3 strain. These enzymes are key to polysaccharide synthesis. By knocking out and complementing the orf1595 and cps4I genes, combined with transcriptome analysis, we preliminarily determined their functions. The results showed that EPS yield decreased significantly after the gene knockout but was restored after complementation. Transcriptome sequencing revealed that knockout of these glycosyltransferase genes led to differential expression of genes within the EPS synthesis cluster and affected sugar metabolism pathways. Furthermore, EPS antioxidant activity was significantly altered: DPPH radical and superoxide anion scavenging abilities decreased, while hydroxyl radical scavenging increased. Chemical analysis indicated changes in EPS protein, glucuronic acid, and sulfate contents after knockout. This study clarifies the critical roles of orf1595 and cps4I in EPS biosynthesis by L. plantarum YM-4-3, providing a theoretical basis for regulating EPS synthesis and facilitating the development of LAB EPS with diverse physiological activities.

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来源期刊

Carbohydrate Research 化学-生化与分子生物学

CiteScore

5.00

自引率

3.20%

发文量

183

审稿时长

3.6 weeks

期刊介绍： Carbohydrate Research publishes reports of original research in the following areas of carbohydrate science: action of enzymes, analytical chemistry, biochemistry (biosynthesis, degradation, structural and functional biochemistry, conformation, molecular recognition, enzyme mechanisms, carbohydrate-processing enzymes, including glycosidases and glycosyltransferases), chemical synthesis, isolation of natural products, physicochemical studies, reactions and their mechanisms, the study of structures and stereochemistry, and technological aspects. Papers on polysaccharides should have a "molecular" component; that is a paper on new or modified polysaccharides should include structural information and characterization in addition to the usual studies of rheological properties and the like. A paper on a new, naturally occurring polysaccharide should include structural information, defining monosaccharide components and linkage sequence. Papers devoted wholly or partly to X-ray crystallographic studies, or to computational aspects (molecular mechanics or molecular orbital calculations, simulations via molecular dynamics), will be considered if they meet certain criteria. For computational papers the requirements are that the methods used be specified in sufficient detail to permit replication of the results, and that the conclusions be shown to have relevance to experimental observations - the authors'' own data or data from the literature. Specific directions for the presentation of X-ray data are given below under Results and "discussion".