Summary
Analysis of the enzymes involved in the biosynthesis of alginic acid by mucoidPseudomonas aeruginosa PAO strain's determined the presence of enzymes required to synthesise GDP-mannuronic acid. Addition of polymannuronic acid to an ammonium sulphate precipitate of a cell free alginate suspension indicated the presence of an enzyme which catalysed the epimerisation of mannuronic acid to guluronic acidafter the polymer had been synthesised. The epimerase was shown to be calcium dependant.
Various non-mucoid mutants were also studied. The non-mucoid parental strain PAO 381 also contained the enzymes required for alginate synthesis but they were not expressed. Synthesis of alginic acid led to an increase in the level of these enzymes. In the non-mucoid mutants derived from mucoid parents GDP-mannose dehydrogenase was absent in all strains studied. In some of these strains GDP-mannose pyrophosphorylase was also absent, while in other strains, phosphomannase isomerase was absent or greatly reduced.
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References
Couperwhite I, McCallum MF (1975) Polysaccharide production and the possible occurence of GDP-Mannose dehydrogenase inAzotobacter vinelandii. Antonie van Leeuwenhoek 41: 25–32
Davidson IW, Sutherland IW, Lawson CJ (1977) Localisation of O-acetyl groups in bacterial alginate. J Gen Microbiol 98: 606–606
Fargie B, Holloway B (1965) Absence of clustering for functionally releated genes inPs. aeruginosa. Genet Res 6:284–299
Frannson LA, Malstrom A, Lindahl U, Hook M (1973) Biosynthesis of dermatan sulphate in fibroblasts. In: Kulonen E, Pikkatainen J (eds) Biology of the Fibroblasts, Academic Press, New York London pp 439–448
Fyfe JAM, Govan JRW (1978) A genetic approach to the study of mucoidPseudomonas aeruginosa. Proc Soc Gen Microbiol 5:54
Fyfe JAM, Govan JRW (1980) Alginate synthesis by mucoidPseudomonas aeruginosa: a chromosomal locus involved in control. J Gen Microbiol 119:443–450
Gorin PAJ, Spencer JFT (1965) Exocellular alginic acid fromAzotobacter vinelandii. Can J Chem 44:993–998
Haug A, Larsen B (1970) Biosynthesis of alginate. Part II. Polymannuronic C-5 epimerase fromAzotobacter vinelandii (Lipmann). Carbohydrate Res 17:287–308
Hook M, Lindahl U, Iverus PH (1974) Distribution of sulphate and iduronic acid residues in Heparin and Heparan sulphate. Biochem J 137:33–43
Kang S, Markovitz A (1967) Induction of capsular polysaccharide synthesis by fluoro-phenylalanine inE. coli wt and strains with altered phenylalanyl soluble ribonucleic acid synthertase. J Bacteriol 93:584–591
Larsen B, Haug (1970) Biosynthesis of alginate. Part 1. Composition and structure of alginate produced byAzotobacter vinelandii (Lipman). Carbohydrate Res 17:287–296
Larsen B, Haug A (1971) Biosynthesis of alginate. Part III. Tritium incorporation with polymannuronic acid C-5 epimerase fromAzotobacter vinelandii. Carbohydrate Res 20:225–232
Lin TY, Hassid WZ (1966a) Isolation of guanosine diphosphate uronic acids from a marine brown alga,Fucus gardneri silva. J Biol Chem 241:3282–3293
Lin TY, Hassid WZ (1966b) Pathway of alginic acid synthesis in the marine brown alga.Fucus gardneri silva. J Biol Chem 241:5284–5297
Linker S, Jones RS (1964) A polysaccharide resembling alginic acid from aPseudomonas micro organism. Nature 204:187–188
Lowry OH, Rosenbrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin-phenol reagent. J Biol Chem 193:265–275
Madgwick J, Haug A, Larsen B (1973) Polymannuronic acid 5-epimerase from the marine algaPelvetia canaliculata (L) Dene et Thur. Acta Chem Scanad 9:3592–3594
Munch-Petersen A (1962) GDPM pyrophosphorylase. In: Colowick SP, Caplan NO (eds) Methods in Enzymology, Vol V. Academic Press, New York London, p 171–174
Ng FM-W, Dawes EA (1973) Chemostat studies on the regulation of glucose metabolism inPs aeruginosa by citrate. Biochem J 132:129–140
Page W, Sadoff HL (1975) Relationship between calcium and uronic acids in encystement ofAzotobacter vinelandii. J Bacteriol 122:145–151
Penman A, Sanderson GR (1972) A method for determination of uronic acid sequences in alginate. Carbohydrate Res 25: 273–282
Pindar DP, Bucke C (1975) The biosynthesis of alginic acid byAzotobacter vinelandii. Biochem J 152:617–622
Preiss J (1964) Sugar nucleotides inArthrobacter. II Biosynthesis of guanosine diphosphomanuronate. J Biol Chem 239:3127–3132
Slein MW (1955) Phosphohexoseisomerases from muscle. Phosphomannose isomerase. In: Colowick SP, Kaplan NO (eds) Methods in Enzymology, Vol 1. Academic Press, New York London, pp 299–304
Sutherland IW, Wilkinson JF (1965) Depolymerases for bacterial polysaccharides obtained from phage infected bacteria. J Gen Microbiol 39:373–378
Weissbach A, Hurwitz J (1958) The formation of 2-keto-3 deoxyheptonic acid in extracts ofE. coli B. J Gen Microbiol 234:705–709
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Piggott, N.H., Sutherland, I.W. & Jarman, T.R. Enzymes involved in the biosynthesis of alginate byPseudomonas aeruginosa . European J. Appl. Microbiol. Biotechnol. 13, 179–183 (1981). https://doi.org/10.1007/BF00703050
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DOI: https://doi.org/10.1007/BF00703050