This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Ye, R W Articles by Chakrabarty, A M Search for Related Content PubMed PubMed Citation Articles by Ye, R W Articles by Chakrabarty, A M

 Previous Article  |  Next Article 

J Bacteriol. 1994 August; 176(16): 4851-4857

research-article

Purification and characterization of phosphomannomutase/phosphoglucomutase from Pseudomonas aeruginosa involved in biosynthesis of both alginate and lipopolysaccharide.

Department of Microbiology and Immunology, University of Illinois, College of Medicine, Chicago 60612.

ABSTRACT

The algC gene from Pseudomonas aeruginosa has been shown to encode phosphomannomutase (PMM), an essential enzyme for biosynthesis of alginate and lipopolysaccharide (LPS). This gene was overexpressed under control of the tac promoter, and the enzyme was purified and its substrate specificity and metal ion effects were characterized. The enzyme was determined to be a monomer with a molecular mass of 50 kDa. The enzyme catalyzed the interconversion of mannose 1-phosphate (M1P) and mannose 6-phosphate, as well as that of glucose 1-phosphate (G1P) and glucose 6-phosphate. The apparent Km values for M1P and G1P were 17 and 22 microM, respectively. On the basis of Kcat/Km ratio, the catalytic efficiency for G1P was about twofold higher than that for M1P. PMM also catalyzed the conversion of ribose 1-phosphate and 2-deoxyglucose 6-phosphate to their corresponding isomers, although activities were much lower. Purified PMM/phosphoglucomutase (PGM) required Mg2+ for maximum activity; Mn2+ was the only other divalent metal that showed some activation. The presence of other divalent metals in addition to Mg2+ in the reaction inhibited the enzymatic activity. PMM and PGM activities could not be detected in nonmucoid algC mutant strain 8858 and in LPS-rough algC mutant strain AK1012, while they were present in the wild-type strains as well as in algC-complemented mutant strains. This evidence suggests that AlgC functions as PMM and PGM in vivo, converting phosphomannose and phosphoglucose in the biosynthesis of both alginate and LPS.

This article has been cited by other articles:

• Yoon, S.-S., Park, S.-H., Kim, Y.-C., Shin, M., Chong, C.-K., Choi, J.-D. (2008). Cloning and Characterization of Phosphoglucomutase and Phosphomannomutase Derived from Sphingomonas chungbukensis DJ77. J Biochem 144: 507-512 [Abstract] [Full Text]  
• Akutsu, J.-i., Zhang, Z., Tsujimura, M., Sasaki, M., Yohda, M., Kawarabayasi, Y. (2005). Characterization of a Thermostable Enzyme with Phosphomannomutase/Phosphoglucomutase Activities from the Hyperthermophilic Archaeon Pyrococcus horikoshii OT3. J Biochem 138: 159-166 [Abstract] [Full Text]  
• Boels, I. C., Kleerebezem, M., de Vos, W. M. (2003). Engineering of Carbon Distribution between Glycolysis and Sugar Nucleotide Biosynthesis in Lactococcus lactis. Appl. Environ. Microbiol. 69: 1129-1135 [Abstract] [Full Text]  
• Franklin, M. J., Ohman, D. E. (2002). Mutant Analysis and Cellular Localization of the AlgI, AlgJ, and AlgF Proteins Required for O Acetylation of Alginate in Pseudomonas aeruginosa. J. Bacteriol. 184: 3000-3007 [Abstract] [Full Text]  
• Nakar, D., Gutnick, D. L. (2001). Analysis of the wee gene cluster responsible for the biosynthesis of the polymeric bioemulsifier from the oil-degrading strain Acinetobacter lwoffii RAG-1. Microbiology 147: 1937-1946 [Abstract] [Full Text]  
• Pier, G. B., Coleman, F., Grout, M., Franklin, M., Ohman, D. E. (2001). Role of Alginate O Acetylation in Resistance of Mucoid Pseudomonas aeruginosa to Opsonic Phagocytosis. Infect. Immun. 69: 1895-1901 [Abstract] [Full Text]  
• Tavares, I. M., Jolly, L., Pompeo, F., Leitão, J. H., Fialho, A. M., Sá-Correia, I., Mengin-Lecreulx, D. (2000). Identification of the Pseudomonas aeruginosa glmM Gene, Encoding Phosphoglucosamine Mutase. J. Bacteriol. 182: 4453-4457 [Abstract] [Full Text]  
• Videira, P. A., Cortes, L. L., Fialho, A. M., Sá-Correia, I. (2000). Identification of the pgmG Gene, Encoding a Bifunctional Protein with Phosphoglucomutase and Phosphomannomutase Activities, in the Gellan Gum-Producing Strain Sphingomonas paucimobilis ATCC 31461. Appl. Environ. Microbiol. 66: 2252-2258 [Abstract] [Full Text]  
• Rocchetta, H. L., Burrows, L. L., Lam, J. S. (1999). Genetics of O-Antigen Biosynthesis in Pseudomonas aeruginosa. Microbiol. Mol. Biol. Rev. 63: 523-553 [Abstract] [Full Text]