Purification and Characterization of Biofilm-Associated EPS Exopolysaccharides from ESKAPE Organisms and Other Pathogens

被引:166
作者
Bales, Patrick M. [1 ]
Renke, Emilija Miljkovic [1 ]
May, Sarah L. [1 ]
Shen, Yang [1 ]
Nelson, Daniel C. [1 ,2 ]
机构
[1] Univ Maryland, Inst Biosci & Biotechnol Res, Rockville, MD USA
[2] Univ Maryland, Dept Vet Med, College Pk, MD 20742 USA
来源
PLOS ONE | 2013年 / 8卷 / 06期
关键词
POLYMERIC SUBSTANCES EPS; PSEUDOMONAS-AERUGINOSA; STAPHYLOCOCCUS-EPIDERMIDIS; CAPSULAR POLYSACCHARIDE; ESCHERICHIA-COLI; ACTINOBACILLUS-ACTINOMYCETEMCOMITANS; BIOCHEMICAL-CHARACTERIZATION; INFECTIOUS-DISEASES; BACTERIAL BIOFILMS; COLANIC ACID;
D O I
10.1371/journal.pone.0067950
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
In bacterial biofilms, high molecular weight, secreted exopolysaccharides can serve as a scaffold to which additional carbohydrates, proteins, lipids, and nucleic acids adhere, forming the matrix of the developing biofilm. Here we report methods to extract and purify high molecular weight (>15 kDa) exopolysaccharides from biofilms of eight human pathogens, including species of Staphylcococcus, Klebsiella, Acinetobacter, Pseudomonas, and a toxigenic strain of Escherichia coli O157:H7. Glycosyl composition analysis indicated a high total mannose content across all strains with P. aeruginosa and A. baumannii exopolysaccharides comprised of 80-90% mannose, K. pneumoniae and S. epidermidis strains containing 40-50% mannose, and E. coli with similar to 10% mannose. Galactose and glucose were also present in all eight strains, usually as the second and third most abundant carbohydrates. N-acetyl-glucosamine and galacturonic acid were found in 6 of 8 strains, while arabinose, fucose, rhamnose, and xylose were found in 5 of 8 strains. For linkage analysis, 33 distinct residue-linkage combinations were detected with the most abundant being mannose-linked moieties, in line with the composition analysis. The exopolysaccharides of two P. aeruginosa strains analyzed were consistent with the Psl carbohydrate, but not Pel or alginate. The S. epidermidis strain had a composition rich in mannose and glucose, which is consistent with the previously described slime associated antigen (SAA) and the extracellular slime substance (ESS), respectively, but no polysaccharide intracellular adhesion (PIA) was detected. The high molecular weight exopolysaccharides from E. coli, K. pneumoniae, and A. baumannii appear to be novel, based on composition and/or ratio analysis of carbohydrates.
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