Invertase and phosphatase of yeast in a phosphate-limited continuous culture

  • Kiyoshi Toda
  • Isamu Yabe
  • Toshihiko Yamagata


Deficiency of inorganic phosphate caused the hyper production of invertase and the derepression of acid phosphatase in a continuous culture ofSaccharomyces carlsbergensis. The specific invertase activity was 40,000 enzyme units per g dry cell weight at a dilution rate lower than 0.05 h−1 with a synthetic glucose medium of which the molecular ratio of KH2PO4 to glucose was less than 0.006. This activity is eight fold higher than in a batch growth and 1.5 fold as much as the highest enzyme activity observed so far in a glucose-limited continuous culture.

For the hyper production of invertase, it is necessary to culture the yeast continuously by keeping the Nyholm's conservative inorganic phosphate concentration at less than 0.2 m mole per g dry weight cell. The derepression of acid phosphatase brought about by phosphate deficiency, was similar in both batch and continuous cultures.


Acid Phosphatase Dilution Rate Continuous Culture Inorganic Phosphate Concentration Cell Weight 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



dilution rate of continuous culture (h−1)


invertase concentration in culture (enzyme unit l−1)


acid phosphatase concentration in culture (enzyme unit l−1)


inorganic phosphate concentration in culture (mM)


glucose concentration in culture (mM)


cell concentration in culture (g dry weight cell l−1)

Greek Letter


specific rate of growth (h−1)





initial value


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  1. Acevedo F, Cooney CL (1973) Penicillin amidase production byBacillus megaterium. Biotechnol Bioeng 15:493–503Google Scholar
  2. Arnold WN (1972) The structure of the yeast cell wall. J Biol Chem 247:1161–1169Google Scholar
  3. Bernt E, Gutmann I (1974) Ethanol determination with alcohol dehydrogenase and NAD. In: Bergmeyer HU (ed) Methods of Enzymatic Analysis, vol 3, 2nd ed. Academic Press Inc, New York, pp 1499–1502Google Scholar
  4. Brown CM (1976) Nitrogen metabolism in bacteria and fungi. In: Dean ACR, Ellwood DC, Evans CGT, Melling J (eds) Continuous culture 6: applications and new fields, Ellis Horwood Ltd, Chichester, pp 170–183Google Scholar
  5. Button DK (1969) Thiamine limited steady state growth of the yeastCryptococcus albidus. J Gen Microbiol 58:15–21Google Scholar
  6. Chen PS Jr, Tribara YT, Warner H (1956) Microdetermination of phosphorus. Anal Chem 28:1756–1758Google Scholar
  7. Dean ACR, Rogers PL (1967) The cell size and macromolecular composition ofAerobacter aerogenes in various systems of continuous culture. Biochim Biophys Acta 148:267–316Google Scholar
  8. Downie JA, Garland PB (1973) An antimycin A- and cyanide-resistant variant ofCandida utilis arising during copper-limited growth. Biochem J 134:1051–1061Google Scholar
  9. Jensen TE, Sicko LM (1974) Phosphate metabolism in blue-green algae. I. Fine structure of the “polyphosphate overplus” phenomenon inPlectonema boryanum. Can J Microbiol 20:1235–1239Google Scholar
  10. Nyholm N (1976) A mathematical model for microbial growth under limitation by conservative substrates. Biotechnol Bioeng 18:1043–1056Google Scholar
  11. Schlanderer G, Dellweg H (1974) Cyclic AMP and catabolite repression in yeasts. InSchizosaccharomyces pombe glucose lowers both intracellular adenosine 3′:5′-monophosphate levels and the activity of catabolite-sensitive enzymes. Eur J Biochem 49:305–316Google Scholar
  12. Schurr A, Yagil E (1971) Regulation and characterization of acid and alkaline phosphatase in yeast. J Gen Microbiol 65:291–303Google Scholar
  13. Slezak J, Sikyta B (1967) Growth ofEscherichia coli B in a continuous culture under limitation by inorganic phosphate. Folia Microbiologica 12:441–446Google Scholar
  14. Smith RW, Dean ACR (1972)β-Galactosidase synthesis inKlebsiella aerogenes growing in continuous culture. J Gen Microbiol 72:37–47Google Scholar
  15. Tempest DW, Dicks JW, Meers JL (1967) Magnesium-limited growth ofBacillus subtilis, in pure and mixed cultures, in a chemostat. J Gen Microbiol 49:139–147Google Scholar
  16. Toda K (1976) Invertase biosynthesis bySaccharomyces carlsbergensis in batch and continuous cultures. Biotechnol Bioeng 18:1103–1115Google Scholar
  17. Toda K, Yabe I (1979a) Mathematical model of cell growth and phosphatase biosynthesis inSaccharomyces carlsbergensis under phosphate limitation. Biotechnol Bioeng 21:487–502Google Scholar
  18. Toda K, Takeuchi T, Sano H (1979b) Growth rate dependence of enzyme synthesis in chemostat culture:α-amylase,β-galactosidase, acid phosphatase andβ-fructosidase. J Chem Tech Biotechnol 29:747–755Google Scholar
  19. Toda K, Yabe I, Yamagata T (1980) Kinetics of biphasic growth of yeast in continuous and fed-batch cultures. Biotechnol Bioeng 22:1805–1827Google Scholar

Copyright information

© Springer-Verlag 1982

Authors and Affiliations

  • Kiyoshi Toda
    • 1
  • Isamu Yabe
    • 1
  • Toshihiko Yamagata
    • 1
  1. 1.Institute of Applied MicrobiologyUniversity of TokyoTokyo

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