Structure and carbohydrate analysis of the exopolysaccharide capsule of Pseudomonas putida G7

被引:83
作者
Kachlany, SC
Levery, SB
Kim, JS
Reuhs, BL
Lion, LW
Ghiorse, WC
机构
[1] Cornell Univ, Dept Microbiol, Ithaca, NY 14853 USA
[2] Univ Georgia, Complex Carbohydrate Res Ctr, Athens, GA 30602 USA
[3] Cornell Univ, Sch Civil & Environm Engn, Ithaca, NY 14850 USA
关键词
D O I
10.1046/j.1462-2920.2001.00248.x
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The aromatic hydrocarbon-degrading bacterium, Pseudomonas putida G7, produces exopolymers of potential interest in biotechnological applications. These exopolymers have been shown to have significant metal-binding ability. To initiate the study of the metal-polymer interactions, we explored the physical and chemical nature of the P. putida G7 exopolysaccharide, a major component of the exo-polymer. A capsular structure was observed by light microscopy surrounding both planktonic and attached cells in biofilms after immunofluorescence staining with polyclonal antiserum raised against planktonic cells. Further work with planktonic cells showed that the immunostained capsule remained associated with young (log phase) cells, whereas older (stationary phase) cells lost their capsular material to the external milieu. Visualization of frozen, hydrated stationary phase cells by cryo-field emission scanning electron microscopy (cryoFESEM) revealed highly preserved extracellular material. In contrast, conventional scanning electron microscopy (SEM) of stationary phase cells showed rope-like material that most probably results from dehydrated and collapsed exopolymer. Both capsular and released exopolymers were separated from cells, and the released extracellular polysaccharide (EPS) was purified. Deoxycholate-polyacrylamide gel electrophoresis (PAGE) and silver/alcian blue staining of the partially purified material showed that it contained both EPS and lipopolysaccharide (LPS). Further purification of the EPS using a differential solubilization technique to remove LPS yielded highly purified EPS. Gas chromatography-mass spectrometry revealed that the purified EPS contained the monosaccharides, glucose, rhamnose, ribose, N-acetylgalactosamine and glucuronic acid. The structural and chemical properties of the P. putida EPS described here increase our understanding of the mechanisms of toxic metal binding by this well-known Proteobacterium.
引用
收藏
页码:774 / 784
页数:11
相关论文
共 56 条
[11]   MOBILIZATION OF ADSORBED CADMIUM AND LEAD IN AQUIFER MATERIAL BY BACTERIAL EXTRACELLULAR POLYMERS [J].
CHEN, JH ;
LION, LW ;
GHIORSE, WC ;
SHULER, ML .
WATER RESEARCH, 1995, 29 (02) :421-430
[12]  
COOKSEY KE, 1992, EXTRACELLULAR POLYM
[13]   MICROBIAL BIOFILMS [J].
COSTERTON, JW ;
LEWANDOWSKI, Z ;
CALDWELL, DE ;
KORBER, DR ;
LAPPINSCOTT, HM .
ANNUAL REVIEW OF MICROBIOLOGY, 1995, 49 :711-745
[14]   Evaluation of the utility of bacterial extracellular polymers for treatment of metal-contaminated soils: Polymer persistence, mobility, and the influence of lead [J].
Czajka, DR ;
Lion, LW ;
Shuler, ML ;
Ghiorse, WC .
WATER RESEARCH, 1997, 31 (11) :2827-2839
[15]   Exopolysaccharide production is required for development of Escherichia coli K-12 biofilm architecture [J].
Danese, PN ;
Pratt, LA ;
Kolter, R .
JOURNAL OF BACTERIOLOGY, 2000, 182 (12) :3593-3596
[16]   Sorption of plutonium, americium and fission products from reprocessing effluents using Rhizopus arrhizus [J].
Dhami, PS ;
Kannan, R ;
Gopalakrishnan, V ;
Ramanujam, A ;
Salvi, N ;
Udupa, SR .
BIOTECHNOLOGY LETTERS, 1998, 20 (09) :869-872
[17]   SEM IMAGING OF BACTERIA AND NANNOBACTERIA IN CARBONATE SEDIMENTS AND ROCKS [J].
FOLK, RL .
JOURNAL OF SEDIMENTARY PETROLOGY, 1993, 63 (05) :990-999
[18]   Metal binding capabilities of Rhizobium etli and its extracellular polymeric substances [J].
Foster, LJR ;
Moy, YP ;
Rogers, PL .
BIOTECHNOLOGY LETTERS, 2000, 22 (22) :1757-1760
[19]  
GEDDIE JL, 1994, J BACTERIOL, V178, P2186
[20]  
GERHARDT P, 1994, METHODS GEN MOL BACT