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洋葱伯克霍尔德菌复合体用于胞外多糖生物合成的多功能性:一项比较结构研究

Versatility of the Burkholderia cepacia complex for the biosynthesis of exopolysaccharides: a comparative structural investigation.

作者信息

Cuzzi Bruno, Herasimenka Yury, Silipo Alba, Lanzetta Rosa, Liut Gianfranco, Rizzo Roberto, Cescutti Paola

机构信息

Department of Life Sciences, University of Trieste, Trieste, Italy.

Department of Chemical Sciences, University of Naples Federico II, Naples, Italy.

出版信息

PLoS One. 2014 Apr 10;9(4):e94372. doi: 10.1371/journal.pone.0094372. eCollection 2014.

Abstract

The Burkholderia cepacia Complex assembles at least eighteen closely related species that are ubiquitous in nature. Some isolates show beneficial potential for biocontrol, bioremediation and plant growth promotion. On the contrary, other strains are pathogens for plants and immunocompromised individuals, like cystic fibrosis patients. In these subjects, they can cause respiratory tract infections sometimes characterised by fatal outcome. Most of the Burkholderia cepacia Complex species are mucoid when grown on a mannitol rich medium and they also form biofilms, two related characteristics, since polysaccharides are important component of biofilm matrices. Moreover, polysaccharides contribute to bacterial survival in a hostile environment by inhibiting both neutrophils chemotaxis and antimicrobial peptides activity, and by scavenging reactive oxygen species. The ability of these microorganisms to produce exopolysaccharides with different structures is testified by numerous articles in the literature. However, little is known about the type of polysaccharides produced in biofilms and their relationship with those obtained in non-biofilm conditions. The aim of this study was to define the type of exopolysaccharides produced by nine species of the Burkholderia cepacia Complex. Two isolates were then selected to compare the polysaccharides produced on agar plates with those formed in biofilms developed on cellulose membranes. The investigation was conducted using NMR spectroscopy, high performance size exclusion chromatography, and gas chromatography coupled to mass spectrometry. The results showed that the Complex is capable of producing a variety of exopolysaccharides, most often in mixture, and that the most common exopolysaccharide is always cepacian. In addition, two novel polysaccharide structures were determined: one composed of mannose and rhamnose and another containing galactose and glucuronic acid. Comparison of exopolysaccharides obtained from cultures on agar plates with those extracted from biofilms on cellulose membranes showed important differences, thus suggesting that extrapolating data from non-biofilm conditions might not always be applicable.

摘要

洋葱伯克霍尔德菌复合体至少由十八个密切相关的物种组成,它们在自然界中广泛存在。一些分离株在生物防治、生物修复和促进植物生长方面显示出有益的潜力。相反,其他菌株是植物和免疫功能低下个体(如囊性纤维化患者)的病原体。在这些个体中,它们可引起呼吸道感染,有时会导致致命后果。大多数洋葱伯克霍尔德菌复合体物种在富含甘露醇的培养基上生长时呈黏液状,并且它们还会形成生物膜,这两个特征相关,因为多糖是生物膜基质的重要组成部分。此外,多糖通过抑制中性粒细胞趋化性和抗菌肽活性以及清除活性氧来促进细菌在恶劣环境中的存活。文献中的大量文章证实了这些微生物产生具有不同结构的胞外多糖的能力。然而,关于生物膜中产生的多糖类型及其与非生物膜条件下获得的多糖的关系知之甚少。本研究的目的是确定洋葱伯克霍尔德菌复合体九个物种产生的胞外多糖类型。然后选择两个分离株,比较在琼脂平板上产生的多糖与在纤维素膜上形成的生物膜中产生的多糖。使用核磁共振光谱、高效尺寸排阻色谱和气相色谱-质谱联用进行了研究。结果表明,该复合体能够产生多种胞外多糖,且大多为混合物,最常见的胞外多糖始终是洋葱多糖。此外,还确定了两种新的多糖结构:一种由甘露糖和鼠李糖组成,另一种含有半乳糖和葡萄糖醛酸。比较琼脂平板培养物中获得的胞外多糖与从纤维素膜生物膜中提取的胞外多糖,发现了重要差异,因此表明从非生物膜条件下推断数据可能并不总是适用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dc83/3983119/47e223b8daa8/pone.0094372.g001.jpg

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