Abstract
The cultivation of the hyperthermophilic archaeobacterium Pyrococcus woesei on starch under continuous gassing (80% H2:20% CO2) caused the formation of 250 U/l of an extremely thermoactive and thermostable α-amylase. In a complex medium without elemental sulphur under 80% N2 and 20% CO2 atmosphere enzyme production could be elevated up to 1000 U/l. Pyrococcus woesei grew preferentially on poly-and oligosaccharides. The amylolytic enzyme formation was constitutive. Enzyme production was also observed in continuous culture at dilution rates from 0.1 to 0.4 h-1. A 20-fold enrichment of α-amylase was achieved after adsorption of the enzyme onto starch and its desorption by preparative gel electrophoresis. The α-amylase consisted of a single subunit with a molecular mass of 70 000 and was catalytically active at a temperature range between 40°C and 130°C. Enzymatic activity was detected even after autoclaving at a pressure of 2 bars at 120°C for 5 h. The purified enzyme hydrolyzed exclusively α-1,4-glycosidic linkages present in glucose polymers of various sizes. Unlike many α-amylases from anaerobes the enzyme from P. woesei was unable to attack short chain oligosaccharides with a chain length between 2 and 6 glucose units.
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Koch, R., Spreinat, A., Lemke, K. et al. Purification and properties of a hyperthermoactive α-amylase from the archaeobacterium Pyrococcus woesei . Arch. Microbiol. 155, 572–578 (1991). https://doi.org/10.1007/BF00245352
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DOI: https://doi.org/10.1007/BF00245352