1887

Abstract

Summary: was grown in continuous culture (34 °C, pH 6.8) under various conditions with respect to dilution rate, nutrient limitation and carbon substrate. Poly-3-hydroxybutyrate (PHB) content, the rate of PHB production ( ) and the rate of carbon substrate utilization ( ) during growth on glucose were maximum at low dilution rate under ammonia limitation (ammonia limitation > potassium/oxygen limitation > glucose limitation). PHB content decreased in a linear manner as a function of dilution rate, from approximately 80% at 0-025 h during ammonia-limited growth to approximately 5% during growth at the maximum specific growth rate (μ) in batch culture. PHB content, and s varied with the nature of the carbon substrate during ammonia-limited growth at fixed dilution rate, and were maximum during growth on lactate [lactate>pyruvate>glucose/gluconate>fructose; highest 0.38 g PHB (g non-PHB biomass) h]. q was related in an approximately linear manner to the in excess of that required solely for the production of non-PHB biomass. This surplus was higher during growth on lactate than on glucose because was approximately equal to the maximum rate of carbon substrate utilization ( ) during growth on lactate, but much lower than during growth on glucose. The relationship between and surplus was confirmed by the effect of adding formate (as an additional source of NADH and/or ATP) and the uncoupling agent carbonyl cyanide--chlorophenylhydrazone (CCCP) to ammonia-limited cultures. It is concluded that is unable to regulate the rate at which it takes up excess carbon substrate to match that required solely for growth, particularly during growth on lactate at low dilution rate, and thus produces PHB as a means of avoiding the potentially deleterious effects of generating high concentrations of intracellular metabolites. Possible ways of further increasing PHB production are discussed.

Funding
This study was supported by the:
  • Collaborative Awards in Science and Engineering studentship (UK Science and Engineering Research Council)
Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-143-7-2361
1997-07-01
2021-08-03
Loading full text...

Full text loading...

/deliver/fulltext/micro/143/7/mic-143-7-2361.html?itemId=/content/journal/micro/10.1099/00221287-143-7-2361&mimeType=html&fmt=ahah

References

  1. Anderson A. J., Dawes E. A. 1990; Occurrence, metabolism, metabolic role and industrial uses of bacterial poly-hydroyalkanoates. Microbiol Rev 54:450–472
    [Google Scholar]
  2. Bowien B., Windhovel U., Yoo J. G., Bednarski R., Kusian B. 1990; Genetics of CO2 fixation in the chemoautotroph Alcaligenes eutrophus . FEMS Microbiol Rev 87:445–450
    [Google Scholar]
  3. Breuer U., Ackermann J.-U., Babel W. 1995; Accumulation of poly (3-hydroxybutyric acid) and overproduction of exopolysaccharides in a mutant of a methyltrophic bacterium. Can J Microbiol 41 (suppl 1):55–59
    [Google Scholar]
  4. Byrom D. 1987; Polymer synthesis by microorganisms: technology and economics. Trends Biotechnol 5:246–250
    [Google Scholar]
  5. Cornish A., Greenwood J. A., Jones C. W. 1988; The relationship between glucose transport and the production of succinoglucan exopolysaccharide by Agrobacterium radiobacter . J Gen Microbiol 134:3111–3122
    [Google Scholar]
  6. Davidson I. W. 1978; Production of polysaccharide by Xanthomonas campestris in continuous culture. FEMS Microbiol Lett 3:347–349
    [Google Scholar]
  7. Friedbold J., 8t Bowien B. 1993; Physiological and biochemical characterisation of the soluble formate dehydrogenase, a molybdoenzyme from Alcaligenes eutrophus . J Bacteriol 175:4719–4728
    [Google Scholar]
  8. Gutmann I., Wahlefeld A. W. 1974 . In Methods of Enzymatic Analysis , 2nd edn, vol. 3 , pp. 1492–1495 . Edited by Bergmeyer H. U. New York St London: Academic Press;
    [Google Scholar]
  9. Jackson F. A, Dawes E. A. 1976; Regulation of the tricarboxylic acid cycle and poly-β-hydroxybutyrate metabolism in Azotobacter beijerinckii grown under nitrogen or oxygen limitation. J Gen Microbiol 97:303–312
    [Google Scholar]
  10. Karr D. B., Waters J. K., Emerich D. W. 1983; Analysis of poly-β-hydroxybutyrate in Rbizobium japonicum bacteroids by ion-exclusion high-pressure liquid chromatography and UV detection. Appl Environ Microbiol 46:1339–1344
    [Google Scholar]
  11. Lee I., Kim M. K., Chang H. N., Park Y. H. 1995; Regulation of poly-β-hydroxybutyrate synthesis by nicotinamide nucleotide in Alcaligenes eutrophus . FEMS Microbiol Lett 131:35–39
    [Google Scholar]
  12. Linton J. D. 1990; The relationship between metabolite production and the growth efficiency of the producing organism. FEMS Microbiol Rev 75:1–18
    [Google Scholar]
  13. Linton J. D., Evans M., Jones D. S., Gouldney D. N. 1987a; Exocellular succinoglucan production by Agrobacterium radiobacter NCIB 11883. J Gen Microbiol 133:2961–2969
    [Google Scholar]
  14. Linton J. D., Jones D. S., Woodard S. 1987b; Factors that control the rate of exopolysaccharide production by Agrobacterium radiobacter NCIB 11883. J Gen Microbiol 133:2979–2987
    [Google Scholar]
  15. Mansfield D. A., Anderson A. J., Naylor L. A. 1995; Regulation of PHB metabolism in Alcaligenes eutrophus . Can J Microbiol 41 (suppl 1):44–49
    [Google Scholar]
  16. Mian F. A., Jarman T. R., Righelato R. C. 1978; Biosynthesis of exopolysaccharide by Pseudomonas aeruginosa . J Bacteriol 134:418–422
    [Google Scholar]
  17. Oeding V., Schlegel H. G. 1973; β-Ketothiolase from Hydro-genomonas eutropha H16 and its significance in the regulation of poly-β-hydroxybutyrate metabolism. Biochem J 134:239–248
    [Google Scholar]
  18. Page W. J. 1995; Bacterial polyhydroxyalkanoates, natural biodegradable plastics with a great future. Can J Microbiol 41 (suppl 1):1–3
    [Google Scholar]
  19. Page W. J., Knosp O. 1989; Hyperproduction of poly-β-hydroxybutyrate during exponential growth of Azotobacter vinelandii UWD. Appl Environ Microbiol 55:1334–1339
    [Google Scholar]
  20. Russell J. B., Cook G. M. 1995; Energetics of bacterial growth; balance of anabolic and catabolic reactions. Microbiol Rev 59:48–62
    [Google Scholar]
  21. Rye A. J., Drozd J. W., Jones C. W., Linton J. D. 1988; Growth efficiency of Xanthomonas campestris in continuous culture. J Gen Microbiol 134:1055–1061
    [Google Scholar]
  22. Senior P. J., Beech G. A., Ritchie G. A. F., Dawes E. A. 1972; The role of oxygen limitation in the formation of poly-β-hydroxybutyrate during batch and continuous culture of Azotobacter beijerinckii . Biochem J 128:1193–1201
    [Google Scholar]
  23. Steinbüchel A. 1991 Polyhydroxyalkanoic acids. . In Biomaterials , pp. 123–213 . Edited by Byrom D. Basingstoke: Macmillan;
    [Google Scholar]
  24. Steinbüchel A., Schlegel H. G. 1989; Excretion of pyruvate by mutants of Alcaligenes eutrophus which are impaired in the accumulation of poly(β-hydroxybutyric acid) (PHB) under conditions permitting synthesis of PHB. Appl Microbiol Biotechnol 31:168–175
    [Google Scholar]
  25. Williams S. G., Greenwood J. A., Jones C. W. 1996; Physiological and biochemical changes accompanying the loss of mucoidy by Pseudomonas aeruginosa . Microbiology 142:881–888
    [Google Scholar]
  26. Yabuuchi E., Kosako Y., Yano I., Hotta H., Nishiuchi Y. 1995; Transfer of two Burkholderia and an Alcaligenes species to Ralstonia gen. nov.; proposal of Ralstonia pickettii (Ralston, Palleroni and Doudoroff 1973) comb, nov., Ralstonia solanacearum (Smith 1896) comb. nov. and Ralstonia eutropha (Davis 1969) comb. nov. Microbiol Immunol 39:897–904
    [Google Scholar]
  27. Yamane T. 1993; Yield of poly-d( – )-3-hydroxybutyrate from various carbon sources: a theoretical study. Biotechnol Bioeng 41:165–170
    [Google Scholar]
  28. Yamane T., Fukunaga M., Lee Y. W. 1996; Increased PHB productivity by high-cell-density fed-batch culture of Alcaligenes latus, a growth-associated PHB producer. Biotechnol Bioeng 50:197–202
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-143-7-2361
Loading
/content/journal/micro/10.1099/00221287-143-7-2361
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error