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植物乳杆菌胞外多糖生物合成基因簇的转录分析

Transcriptional analysis of exopolysaccharides biosynthesis gene clusters in Lactobacillus plantarum.

作者信息

Vastano Valeria, Perrone Filomena, Marasco Rosangela, Sacco Margherita, Muscariello Lidia

机构信息

Dipartimento di Scienze e Tecnologie Ambientali, Biologiche e Farmaceutiche, Seconda Università di Napoli, via Vivaldi 43, 81100, Caserta, Italy.

出版信息

Arch Microbiol. 2016 Apr;198(3):295-300. doi: 10.1007/s00203-015-1169-1. Epub 2015 Nov 7.

Abstract

Exopolysaccharides (EPS) from lactic acid bacteria contribute to specific rheology and texture of fermented milk products and find applications also in non-dairy foods and in therapeutics. Recently, four clusters of genes (cps) associated with surface polysaccharide production have been identified in Lactobacillus plantarum WCFS1, a probiotic and food-associated lactobacillus. These clusters are involved in cell surface architecture and probably in release and/or exposure of immunomodulating bacterial molecules. Here we show a transcriptional analysis of these clusters. Indeed, RT-PCR experiments revealed that the cps loci are organized in five operons. Moreover, by reverse transcription-qPCR analysis performed on L. plantarum WCFS1 (wild type) and WCFS1-2 (ΔccpA), we demonstrated that expression of three cps clusters is under the control of the global regulator CcpA. These results, together with the identification of putative CcpA target sequences (catabolite responsive element CRE) in the regulatory region of four out of five transcriptional units, strongly suggest for the first time a role of the master regulator CcpA in EPS gene transcription among lactobacilli.

摘要

来自乳酸菌的胞外多糖(EPS)有助于发酵乳制品形成特定的流变学特性和质地,并且在非乳制品食品及治疗学领域也有应用。最近,在植物乳杆菌WCFS1(一种益生菌及与食品相关的乳杆菌)中鉴定出了四组与表面多糖产生相关的基因簇(cps)。这些基因簇参与细胞表面结构的形成,可能还与免疫调节性细菌分子的释放和/或暴露有关。在此,我们展示了对这些基因簇的转录分析。事实上,逆转录-聚合酶链反应(RT-PCR)实验表明,cps基因座由五个操纵子组成。此外,通过对植物乳杆菌WCFS1(野生型)和WCFS1-2(ΔccpA)进行逆转录定量PCR分析,我们证明了三个cps基因簇的表达受全局调节因子CcpA的控制。这些结果,连同在五个转录单元中的四个的调控区域中鉴定出假定的CcpA靶序列(分解代谢物反应元件CRE),首次有力地表明了主调节因子CcpA在乳杆菌中EPS基因转录中的作用。

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