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调控霍乱弧菌黏附颗粒淀粉相关机制的研究。

Mechanisms involved in governing adherence of Vibrio cholerae to granular starch.

机构信息

Department of Biotechnology and Food Engineering, Technion, Haifa 32000, Israel.

出版信息

Appl Environ Microbiol. 2010 Feb;76(4):1034-43. doi: 10.1128/AEM.01533-09. Epub 2009 Dec 18.

Abstract

Vibrio cholerae has been shown to adhere to cornstarch granules. The present work explored the mechanisms involved in this adhesion and the possibility of its occurrence in vivo. The findings suggest that both specific and nonspecific interactions are involved in the adhesion. Nonspecific hydrophobic interactions may play a role, since both V. cholerae and cornstarch granules exhibited hydrophobic properties when they were tested using a xylene-water system. In addition, the presence of bile acids reduced the adhesion. The adhesion also involves some specific carbohydrate-binding moieties on the cell surface, as reflected by reduced adhesion following pretreatment of the bacteria with proteinase K and sodium m-periodate. Further investigations supported these observations and showed that media containing low-molecular-weight carbohydrates had a significant inhibitory effect. Binding cell lysate to starch granules and removing the adhered proteins using either glucose or bile acids led to identification (by liquid chromatography-tandem mass spectrometry analysis) of several candidate V. cholerae outer membrane-associated starch-binding proteins. Different sets of proteins were isolated by removal in a glucose solution or bile acids. When the upper gastrointestinal tract conditions were simulated in vitro, both bile salts and the amylolytic activity of the pancreatic juices were found to have an inhibitory effect on the adherence of V. cholerae to starch. However, during acute diarrhea, this inhibitory effect may be significantly reduced due to dilution, suggesting that adhesion does occur in vivo. Such adhesion may contribute to the beneficial effects observed following administration of granular starch-based oral rehydration solutions to cholera patients.

摘要

霍乱弧菌已被证明能黏附在玉米淀粉颗粒上。本研究探索了这种黏附的机制以及其在体内发生的可能性。研究结果表明,特定和非特定相互作用都参与了黏附。非特异性疏水性相互作用可能起作用,因为当使用二甲苯-水体系测试时,霍乱弧菌和玉米淀粉颗粒都表现出疏水性。此外,胆汁酸的存在减少了黏附。黏附还涉及细胞表面上的一些特定的碳水化合物结合位点,因为用蛋白酶 K 和高碘酸钠预处理细菌后,黏附减少。进一步的研究支持了这些观察结果,并表明含有低分子量碳水化合物的培养基具有显著的抑制作用。将细胞裂解物结合到淀粉颗粒上,并使用葡萄糖或胆汁酸去除黏附的蛋白质,导致鉴定出几种候选霍乱弧菌外膜相关的淀粉结合蛋白(通过液相色谱-串联质谱分析)。用葡萄糖溶液或胆汁酸去除可分离出不同的蛋白质组。当在体外模拟上消化道条件时,发现胆汁盐和胰液的淀粉酶活性对霍乱弧菌黏附淀粉都有抑制作用。然而,在急性腹泻期间,由于稀释,这种抑制作用可能会显著降低,表明体内确实会发生黏附。这种黏附可能有助于观察到颗粒状淀粉基口服补液溶液对霍乱患者的有益效果。

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