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Microbiology 152 (2006), 1807-1818; DOI  10.1099/mic.0.28611-0
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Microbiology 152 (2006), 1807-1818; DOI  10.1099/mic.0.28611-0
© 2006 Society for General Microbiology

The ionic interaction of Klebsiella pneumoniae K2 capsule and core lipopolysaccharide

Sandra Fresno1, Natalia Jiménez1, Luis Izquierdo1, Susana Merino1, Maria Michela Corsaro3, Cristina De Castro3, Michelangelo Parrilli3, Teresa Naldi3, Miguel Regué2 and Juan M. Tomás1

1 Departamento de Microbiología, Facultad de Biología, Universidad de Barcelona, Diagonal 645, 08071 Barcelona, Spain
2 Departamento de Microbiología y Parasitología Sanitarias, Facultad de Farmacia, Universidad de Barcelona, Av. Joan XXIII s/n, 08028 Barcelona, Spain
3 Dipartimento di Chimica e Biochimica, Università Federico II di Napoli, Complesso Universitario Monte S. Angelo, Via Cintia 4, 80126 Napoli, Italy

Correspondence
Juan M. Tomás
jtomas{at}ub.edu

The complete structures of LPS core types 1 and 2 from Klebsiella pneumoniae have been described by other authors. They are characterized by a lack of phosphoryl residues, but they contain galacturonic acid (GalA) residues, which contribute to the necessary negative charges. The presence of a capsule was determined in core-LPS non-polar mutants from strains 52145 (O1 : K2), DL1 (O1 : K1) and C3 (O8 : K66). O-antigen ligase (waaL) mutants produced a capsule. Core mutants containing the GalA residues were capsulated, while those lacking the residues were non capsulated. Since the proteins involved in the transfer of GalA (WabG) and glucosamine residues (WabH) are known, the chemical basis of the capsular-K2–cell-surface association was studied. Phenol/water extracts from K. pneumoniae 52145{Delta}wabH waaL and 52145{Delta}waaL mutants, but not those from from K. pneumoniae 52145{Delta}wabG waaL mutant, contained both LPS and capsular polysaccharide, even after hydrophobic chromatography. The two polysaccharides were dissociated by gel-filtration chromatography, eluting with detergent and metal-ion chelators. From these results, it is concluded that the K2 capsular polysaccharide is associated by an ionic interaction to the LPS through the negative charge provided by the carboxyl groups of the GalA residues.

Abbreviations: DOC, deoxycholate; EM, electron microscopy; GalA, galacturonic acid; Glc, D-glucopyranose; GlcN, D-glucosamine; Hep, L-glycero-D-manno-heptopyranose; Kdo, 3-deoxy-D-manno-oct-2-ulosonic acid; K-PS, capsular polysaccharide; MALDI-TOF, matrix-assisted laser desorption ionization time-of-flight; NaOAc, sodium acetate; OM, outer membrane; OMP, outer-membrane protein; O-PS, O-antigen polysaccharide; OS, oligosaccharide(s); PCP, phenol/chloroform/light petroleum ether; PW, phenol/water




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