Skip to main content
NCBI home page
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1989 Mar;55(3):689–694. doi: 10.1128/aem.55.3.689-694.1989

Continuous Production of Thermostable β-Amylase with Clostridium thermosulfurogenes: Effect of Culture Conditions and Metabolite Levels on Enzyme Synthesis and Activity

A Nipkow 1, G-J Shen 1, J G Zeikus 1,*
PMCID: PMC184181  PMID: 16347874

Abstract

A β-amylase-overproducing mutant of Clostridium thermosulfurogenes was grown in continuous culture on soluble starch to produce thermostable β-amylase. Enzyme productivity was reasonably stable over periods of weeks to months. The pH and temperature optima for β-amylase production were pH 6.0 and 60°C, respectively. Enzyme concentration was maximized by increasing biomass concentration by using high substrate concentrations and by maintaining a low growth rate. β-Amylase concentration reached 90 U ml−1 at a dilution rate of 0.07 h−1 in a 3% starch medium. A further increase in enzyme activity levels was limited by acetic acid inhibition of growth and low β-amylase productivity at low growth rates.

Full text

PDF
689

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Baronofsky J. J., Schreurs W. J., Kashket E. R. Uncoupling by Acetic Acid Limits Growth of and Acetogenesis by Clostridium thermoaceticum. Appl Environ Microbiol. 1984 Dec;48(6):1134–1139. doi: 10.1128/aem.48.6.1134-1139.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Davis P. E., Cohen D. L., Whitaker A. The production of alpha-amylase in batch and chemostat culture by Bacillus stearothermophilus. Antonie Van Leeuwenhoek. 1980;46(4):391–398. doi: 10.1007/BF00421985. [DOI] [PubMed] [Google Scholar]
  3. Fabián J. Synthesis of the extracellular protease by Bacillus pumilus. Folia Microbiol (Praha) 1970;15(3):160–168. doi: 10.1007/BF02873079. [DOI] [PubMed] [Google Scholar]
  4. Fencl Z., Pazlarová J. Production of extracellular enzymes in continuous culture. Folia Microbiol (Praha) 1982;27(5):340–349. doi: 10.1007/BF02883137. [DOI] [PubMed] [Google Scholar]
  5. Heineken F. G., O'Connor R. J. Continuous culture studies on the biosynthesis of alkaline protease, neutral protease and -amylase by Bacillus subtilis NRRL-B3411. J Gen Microbiol. 1972 Nov;73(1):35–44. doi: 10.1099/00221287-73-1-35. [DOI] [PubMed] [Google Scholar]
  6. Herrero A. A., Gomez R. F. Development of ethanol tolerance in Clostridium thermocellum: effect of growth temperature. Appl Environ Microbiol. 1980 Sep;40(3):571–577. doi: 10.1128/aem.40.3.571-577.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hyun H. H., Zeikus J. G. General Biochemical Characterization of Thermostable Extracellular beta-Amylase from Clostridium thermosulfurogenes. Appl Environ Microbiol. 1985 May;49(5):1162–1167. doi: 10.1128/aem.49.5.1162-1167.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hyun H. H., Zeikus J. G. Regulation and genetic enhancement of beta-amylase production in Clostridium thermosulfurogenes. J Bacteriol. 1985 Dec;164(3):1162–1170. doi: 10.1128/jb.164.3.1162-1170.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hyun H. H., Zeikus J. G. Simultaneous and Enhanced Production of Thermostable Amylases and Ethanol from Starch by Cocultures of Clostridium thermosulfurogenes and Clostridium thermohydrosulfuricum. Appl Environ Microbiol. 1985 May;49(5):1174–1181. doi: 10.1128/aem.49.5.1174-1181.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Jensen D. E. Continuous production of extracellular protease by Bacillus subtilis in a two-stage fermentor. Biotechnol Bioeng. 1972 Jul;14(4):647–662. doi: 10.1002/bit.260140410. [DOI] [PubMed] [Google Scholar]
  11. Lovitt R. W., Longin R., Zeikus J. G. Ethanol Production by Thermophilic Bacteria: Physiological Comparison of Solvent Effects on Parent and Alcohol-Tolerant Strains of Clostridium thermohydrosulfuricum. Appl Environ Microbiol. 1984 Jul;48(1):171–177. doi: 10.1128/aem.48.1.171-177.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Schwartz R. D., Keller F. A. Acetic Acid Production by Clostridium thermoaceticum in pH-Controlled Batch Fermentations at Acidic pH. Appl Environ Microbiol. 1982 Jun;43(6):1385–1392. doi: 10.1128/aem.43.6.1385-1392.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. WOLIN E. A., WOLIN M. J., WOLFE R. S. FORMATION OF METHANE BY BACTERIAL EXTRACTS. J Biol Chem. 1963 Aug;238:2882–2886. [PubMed] [Google Scholar]
  14. Wang G., Wang D. I. Elucidation of Growth Inhibition and Acetic Acid Production by Clostridium thermoaceticum. Appl Environ Microbiol. 1984 Feb;47(2):294–298. doi: 10.1128/aem.47.2.294-298.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES