Skip to main content
Log in

Highly active and thermostable amylases and pullulanases from various anaerobic thermophiles

  • Applied Microbiology
  • Published:
Applied Microbiology and Biotechnology Aims and scope Submit manuscript

Summary

High concentrations of amylases and pullulanases were formed by continuous cultivation of Thermoanaerobacter finnii, Thermobacteroides acetoethylicus, Thermoanaerobacter ethanolicus and Clostridium thermosaccharolyticum in chemostats under starch limitation. 70% to 98% of these enzymes were transported and released into the culture fluid. These extracellular enzymes were extremely thermostable under aerobic conditions and in the absence of substrate and metal ions. The amylases and pullulanases from the first three organisms had an optimal temperature of 90°C. The enzymes from C. thermosaccharolyticum were most active at 75°C. The pH optima of the amylolytic enzymes from the microorganisms investigated ranged between 5 and 6. The addition of calcium ions in vitro significantly enhanced pullulanase activity from T. finnii and C. thermosaccharolyticum. The influence of other metal ions and cyclodextrins on the activities of the amylolytic enzymes is also described.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Antranikian G, Gottschalk G (1987) Production of thermostable enzymes for starch hydrolysis. 4th European Congress of Biotechnology. Amsterdam, The Netherlands, pp 1–4

  • Antranikian G, Gottschalk G (1986) German Patent No. P 3 639267.7

  • Antranikian G, Zablowski, Gottschalk G (1987) Conditions for the overproduction and excretion of thermostable glucoamylase and pullulanase from Clostridium thermohydrosulfuricum DSM 567. Appl Microbiol Biotechnol 27:75–81

    Google Scholar 

  • Antranikian G, Herzberg C, Gottschalk G (1987a) Production of thermostable α-amylase, pullulanase and α-glucosidase in continuous culture by a new Clostridium isolate. Appl Environ Microbiol, 53:1668–1673

    Google Scholar 

  • Antranikian G, Herzberg C, Mayer F, Gottschalk G (1987b) Changes in cell-envelope structure of Clostridium strain EM 1 during massive production of amylase and pullulanase. FEMS Lett 41:193–197

    Google Scholar 

  • Bender H, Wallenfels K (1966) Pullulanase (an amylopectin and glycogen debranching enzyme) from Aerobacter aerogenes. In: Neufeld EF, Ginsburg (eds) Method in enzymology VIII. Academic Press, New York, pp 555–562

    Google Scholar 

  • Bergmeyer HU, Gawehn K, Grabe M (1974) Enzyme als biochemische Reagentien. In: Bergmeyer HU (ed) Methoden der Enzymatischen Analyse. Verlag Chemie, Weinheim, pp 454–558

    Google Scholar 

  • Bergmeyer HU, Grassl M (1983) Reagents for enzymatic analysis; enzyme-α-amylase. In: Bergmeyer HU (ed) Methods of enzymatic analysis — II. 3rd ed, Verlag Chemie, Weinheim, FRG, pp 151–152

    Google Scholar 

  • De Mot R, Andries K, Verachtert H (1984a) Comparative study of starch degradation and amylase production by ascomycetous yeast species. Syst Appl Microbiol 5:106–118

    Google Scholar 

  • De Mot R, van Oudendijck E, Verachtert H (1984b) Production of extracellular debranching activity by amylolytic yeasts. Biotechnol Lett 6:581–586

    Google Scholar 

  • Enevoldsen BS, Reimann L, Hansen NL (1977) Biospecific affinity chromatography of pullulanase. FEBS Lett 79:121–124

    Google Scholar 

  • Hyun HH, Zeikus JG (1985a) General biochemical characterization of thermostable pullulanase and glucoamylase from Clostridium thermohydrosulfuricum. Appl Environ Microbiol 49:1168–1173

    Google Scholar 

  • Hyun HH, Zeikus JG (1985b) Simultaneous and enhanced production of thermostable amylase and ethanol from starch by cocultures of Clostridium thermosulfurogenes and Clostridium thermohydrosulfuricum. Appl Environ Microbiol 49:1174–1181

    Google Scholar 

  • Jensen BF, Norman BE (1984) Bacillus acidopullulyticus pullulanase-application and regulatory aspects for use in the food industry. Proc Biochem 19:129–134

    Google Scholar 

  • Konishi Y, Amemura A, Tanabe S, Harada T (1979) Immunological study of pullulanase from Klebsiella strains and the occurrence of this enzyme in the Enterobacteriaceae. Int J Syst Bacteriol 29:13–18

    Google Scholar 

  • Madi E, Antranikian G, Ohmiya K, Gottschalk G (1987) Thermostable amylolytic enzymes from a new Clostridium isolate. Appl Environ Microbiol, in press

  • Nakamura N, Watanabe K, Horikoshi K (1975) Purification and some properties of alkaline pullulanases from a strain of Bacillus No. 202-1, an, alkalophilic microorganism. Biochem Biophys Acta 379:188–193

    Google Scholar 

  • Plant AR, Morgan HW, Daniel RM (1986) A highly stable pullulanase from Thermus aquaticus YT-1. Enzyme Microb Technol 8:668–672

    Google Scholar 

  • Suzuki Y, Chishiro M (1983) Production of extracellular thermostable pullulanase by an amylolytic obligately thermophilic soil bacterium, Bacillus stearothermophilus. K.P. 1064. Eur J Appl Biotechnol 17:24–29

    Google Scholar 

  • Takasaki Y (1976a) Productions and utilizations of β-amylase and pullulanase from Bacillus cereus var. mycoides. Agr Biol Chem 40:1515–1522

    Google Scholar 

  • Takasaki Y (1976b) Purification and enzymatic properties of β-amylase and pullulanase from Bacillus cereus var. mycoides. Agr Biol Chem 40:1523–1530

    Google Scholar 

  • Takizawa N, Murooka Y (1985) Cloning of the pullulanase gene in Escherichia coli and Klebsiella aerogenes. Appl Environ Microbiol 49:294–298

    Google Scholar 

  • Ueda S, Nanri N (1967) Production of isoamylase by Escherichia intermedia. Appl Environ Microbiol 15:492–496

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Koch, R., Zablowski, P. & Antranikian, G. Highly active and thermostable amylases and pullulanases from various anaerobic thermophiles. Appl Microbiol Biotechnol 27, 192–198 (1987). https://doi.org/10.1007/BF00251944

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00251944

Keywords

Navigation