Dataset Information

Relationship Between Putative eps Genes and Production of Exopolysaccharide in Lactobacillus casei LC2W.

ABSTRACT: Lactobacillus casei LC2W, a probiotic strain, can produce exopolysaccharide (EPS) with anti-hypertensive bioactivity. The relationship between eps genes and EPS synthesis in LC2W due to unclear regulation mechanism of EPS biosynthesis was investigated. The several relevant genes in EPS biosynthetic gene cluster were deleted, overexpressed and complemented. The results suggested that glucose-1-phosphate thymidyltranseferase gene (LC2W_2179), uncharacterized EPS biosynthesis protein (LC2W_2188), and EPS biosynthesis protein (LC2W_2189) were related to EPS biosynthesis. EPS titer decreased 15, 13, and 21% when the three genes were deleted, respectively. When they were overexpressed, EPS titer increased 16, 10, and 18%. When they were complemented, EPS titer was similar to the wild-type strain. This work showed the three eps genes from LC2W played important roles on EPS production.

SUBMITTER: Song X

PROVIDER: S-EPMC6107683 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature

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Relationship Between Putative eps Genes and Production of Exopolysaccharide in Lactobacillus casei LC2W.

Song Xin X Xiong Zhiqiang Z Kong Linghui L Wang Guangqiang G Ai Lianzhong L

Frontiers in microbiology 20180817

Lactobacillus casei LC2W, a probiotic strain, can produce exopolysaccharide (EPS) with anti-hypertensive bioactivity. The relationship between eps genes and EPS synthesis in LC2W due to unclear regulation mechanism of EPS biosynthesis was investigated. The several relevant genes in EPS biosynthetic gene cluster were deleted, overexpressed and complemented. The results suggested that glucose-1-phosphate thymidyltranseferase gene (LC2W_2179), uncharacterized EPS biosynthesis p ...[more]

PMID: 30174661

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Project description:Lactobacillus johnsonii FI9785 makes two capsular exopolysaccharides-a heteropolysaccharide (EPS2) encoded by the eps operon and a branched glucan homopolysaccharide (EPS1). The homopolysaccharide is synthesized in the absence of sucrose, and there are no typical glucansucrase genes in the genome. Quantitative proteomics was used to compare the wild type to a mutant where EPS production was reduced to attempt to identify proteins associated with EPS1 biosynthesis. A putative bactoprenol glycosyltransferase, FI9785_242 (242), was less abundant in the Δeps_cluster mutant strain than in the wild type. Nuclear magnetic resonance (NMR) analysis of isolated EPS showed that deletion of the FI9785_242 gene (242) prevented the accumulation of EPS1, without affecting EPS2 synthesis, while plasmid complementation restored EPS1 production. The deletion of 242 also produced a slow-growth phenotype, which could be rescued by complementation. 242 shows amino acid homology to bactoprenol glycosyltransferase GtrB, involved in O-antigen glycosylation, while in silico analysis of the neighboring gene 241 suggested that it encodes a putative flippase with homology to the GtrA superfamily. Deletion of 241 also prevented production of EPS1 and again caused a slow-growth phenotype, while plasmid complementation reinstated EPS1 synthesis. Both genes are highly conserved in L. johnsonii strains isolated from different environments. These results suggest that there may be a novel mechanism for homopolysaccharide synthesis in the Gram-positive L. johnsonii IMPORTANCE Exopolysaccharides are key components of the surfaces of their bacterial producers, contributing to protection, microbial and host interactions, and even virulence. They also have significant applications in industry, and understanding their biosynthetic mechanisms may allow improved production of novel and valuable polymers. Four categories of bacterial exopolysaccharide biosynthesis have been described in detail, but novel enzymes and glycosylation mechanisms are still being described. Our findings that a putative bactoprenol glycosyltransferase and flippase are essential to homopolysaccharide biosynthesis in Lactobacillus johnsonii FI9785 indicate that there may be an alternative mechanism of glucan biosynthesis to the glucansucrase pathway. Disturbance of this synthesis leads to a slow-growth phenotype. Further elucidation of this biosynthesis may give insight into exopolysaccharide production and its impact on the bacterial cell.

Dataset Information

Relationship Between Putative eps Genes and Production of Exopolysaccharide in Lactobacillus casei LC2W.

Publications

Relationship Between Putative <i>eps</i> Genes and Production of Exopolysaccharide in <i>Lactobacillus casei</i> LC2W.

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