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Enzymatic synthesis of poly-β-hydroxybutyrate inZoogloea ramigera


The enzyme activity synthesizing poly-β-hydroxybutyrate (PHB) was mainly localized in the PHB-containing particulate fraction ofZoogloea ramigera I-16-M, when it grew flocculatedly in a medium supplemented with glucose. On the other hand, the enzyme activity remained in the soluble fraction, when the bacterium grew dispersedly in a glucose-starved medium.

The soluble PHB synthase activity became associated with the particulate fraction as PHB synthesis was initiated on the addition of glucose to the dispersed culture. Conversely, the enzyme activity was released from the PHB-containing granules to the soluble fraction when the flocculated culture was kept incubated without supplementing the medium with glucose.

PHB synthase was also incorporated into the newly formed PHB fraction when partially purified soluble PHB synthase was incubated withd(-)-β-hydroxybutyryl CoA in vitro.

Although attempts to solubilize the particulate enzyme were unsuccessful, and the soluble enzyme became extremely unstable in advanced stages of purification, both PHB synthases had the same strict substrate specificity ford(-)-β-hydroxybutyryl CoA, and showed the same pH optimum at 7.0.

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  1. Butterfield, C. T.: A zooglea-forming bacterium isolated from activated sludge. Publ. Hlth Rep. (Wash.)50, 671–684 (1935)

  2. Crabtree, K., Boyle, W., McCoy, E., Rohlich, G. A.: A mechanism of floc formation byZoogloea ramigera. J. Water Pollut. Control Fed.38, 1968–1980 (1966)

  3. Crabtree, K., McCoy, E.:Zoogloea ramigera Itzigsohn, identification and description. Request for an opinion as to the status of the generic nameZoogloea. Int. J. syst. Bacteriol.17, 1–10 (1967)

  4. Crabtree, K., McCoy, E., Boyle, W. C., Rohlich, G. A.: Isolation, identification, and metabolic role of the sudanophilic granules ofZoogloea ramigera. Appl. Microbiol.13, 218–226 (1965)

  5. Dawes, E. A., Ribbons, D. W.: Some aspects of the endogenous metabolism of bacteria. Bact. Rev.28, 126–149 (1964)

  6. Doudoroff, M.: Metabolism of poly-β-hydroxybutyrate in bacteria. In: Current aspects of biochemical energetics (N. O. Kaplan, E. P. Kennedy, eds.), pp. 385–400. New York-London: Academic Press 1966

  7. Friedman, B. A., Dugan, P. R.: Concentration and accumulation of metallic ions by the bacteriumZoogloea. Develop. Ind. Microbiol.9, 381–388 (1968)

  8. Griebel, R. J., Merrick, J. M.: Metabolism of poly-β-hydroxybutyrate: Effect of mild alkaline extraction on native poly-β-hydroxybutyrate granules. J. Bact.108, 782–789 (1971)

  9. Griebel, R. J., Smith, Z., Merrick, J. M.: Metabolism of poly-β-hydroxybutyrate. I. Purification, composition, and properties of native poly-β-hydroxybutyrate granules fromBacillus megaterium. Biochemistry7, 3676–3681 (1968)

  10. Von Korff, R. W.: Ion-exchange chromatography of citric acid cycle components and related compounds. In: Methods in enzymology, Vol. 13 (J. M. Lowenstein, ed.), pp. 425–430. New York-London: Academic Press 1969

  11. Lowry, O. H., Rosebrough, N. J., Farr, A. L., Randall, R. J.: Protein measurement with the Folin phenol reagent. J. biol. Chem.193, 265–275 (1951)

  12. Martin, R. G., Ames, B. N.: A method for determining the sedimentation behavior of enzymes: Application to protein mixtures. J. biol. Chem.236, 1372–1379 (1961)

  13. Merrick, J. M., Doudoroff, M.: Enzymatic synthesis of poly-β-hydroxybutyric acid in bacteria. Nature (Lond.)189, 890–892 (1961)

  14. Senior, P. J., Beech, G. A., Ritchie, G. A. F., Dawes, E. A.: The role of oxygen limitation in the formation of poly-β-hydroxybutyrate during batch and continuous culture ofAzotobacter beijerinckii. Biochem. J.128, 1193–1201 (1972)

  15. Senior, P. J., Dawes, E. A.: The regulation of poly-β-hydroxybutyrate metabolism inAzotobacter beijerinckii. Biochem. J.134, 225–238 (1973)

  16. Slepecky, R. A., Law, J. H.: A rapid spectrophotometric assay of alpha, beta-unsaturated acids and beta-hydroxy acids. Analyt. Chem.32, 1697–1699 (1960)

  17. Stevenson, L. H., Socolofsky, M. D.: Cyst formation and poly-β-hydroxybutyric acid accumulation inAzotobacter. J. Bact.91, 304–310 (1966)

  18. Wieland, T., Rueff, L.: Synthese von S-β-Oxybutyryl- und S-Acetacetyl-Coenzym A. Angew. Chem.65, 186–187 (1953)

  19. Williamson, D. H., Wilkinson, J. F.: The isolation and estimation of the poly-β-hydroxybutyrate inclusions ofBacillus species. J. gen. Microbiol.19, 198–209 (1958)

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Fukui, T., Yoshimoto, A., Matsumoto, M. et al. Enzymatic synthesis of poly-β-hydroxybutyrate inZoogloea ramigera . Arch. Microbiol. 110, 149–156 (1976).

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Key words

  • Poly-β-hydroxybutyrate
  • PHB
  • PHB synthase
  • Zoogloea ramigera