• Title/Summary/Keyword: Sterilization oven's microorganism

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Purification and characterization of a thermostable glutamate dehydrogenase from a thermophilic bacterium isolated from a sterilization drying oven

  • Amenabar, Maximiliano J.;Blamey, Jenny M.
    • BMB Reports
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    • v.45 no.2
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    • pp.91-95
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    • 2012
  • Glutamate dehydrogenase from axenic bacterial cultures of a new microorganism, called GWE1, isolated from the interior of a sterilization drying oven, was purified by anion-exchange and molecular-exclusion liquid chromatography. The apparent molecular mass of the native enzyme was 250.5 kDa and was shown to be an hexamer with similar subunits of molecular mass 40.5 kDa. For glutamate oxidation, the enzyme showed an optimal pH and temperature of 8.0 and $70^{\circ}C$, respectively. In contrast to other glutamate dehydrogenases isolated from bacteria, the enzyme isolated in this study can use both $NAD^+$ and $NADP^+$ as electron acceptors, displaying more affinity for $NADP^+$ than for $NAD^+$. No activity was detected with NADH or NADPH, 2-oxoglutarate and ammonia. The enzyme was exceptionally thermostable, maintaining more than 70% of activity after incubating at $100^{\circ}C$ for more than five hours suggesting being one of the most thermoestable enzymes reported in the family of dehydrogenases.

Characterization of the Thermophilic Bacterium Geobacillus sp. Strain GWE1 Isolated from a Sterilization Oven

  • Correa-Llanten, Daniela;Larrain-Linton, Juanita;Munoz, Patricio A.;Castro, Miguel;Boehmwald, Freddy;Blamey, Jenny M.
    • Microbiology and Biotechnology Letters
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    • v.41 no.3
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    • pp.278-283
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    • 2013
  • A gram-positive, rod-shaped, spore-forming, motile thermophilic bacterium was isolated from a sterilization oven. The microorganism GWE1, formally named Geobacillus wiegelii identified as a member of the genus Geobacillus. GWE1 grew under aerobic conditions of between $60-80^{\circ}C$ (optimum $670^{\circ}C$), in a pH range of 3.0-8.0 (optimum $pH^{70^{\circ}C}$ 5.8), and between 0 and 2 M NaCl (optimum 0.3 M). The membrane polar lipids were dominated by branched saturated fatty acids, which included as the major constituents; iso-15:0 (13.3%), 16:1(${\omega}7$) (12.8%), 16:0 (28.5%), iso-17:0 (13.5%) and anteiso-17:0 (12.3%). The DNA G+C content was 47.2 mol% (determined by HPLC). The 16S rRNA gene sequence of GWE1 showed a high similarity with Geobacillus caldoxylosilyticus (97%). However, the level of DNA-DNA relatedness was only 58%. These data suggest that GWE1 is probably a novel specie of the genus Geobacillus.