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Purification and characterization of an extremely stable glucose isomerase from Geobacillus thermodenitrificans TH2

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Abstract

The D-glucose/D-xylose isomerase was purified from a thermophilic bacterium, Geobacillus thermodenitrificans TH2, by precipitating with heat shock and using Q-Sepharose ion exchange column chromatography, and then characterized. The purified enzyme had a single band having molecular weight of 49 kDa on SDS-PAGE. In the presence of D-glucose as a substrate, the optimum temperature and pH of the enzyme were found to be 80°C and 7.5, respectively. The purified xylose isomerase of G. thermodenitrificans TH2 was extremely stable at pH 7.5 after 96 h incubation at 4°C and 50°C. When the thermal stability profile was analyzed, it was determined that the purified enzyme was extremely stable during incubation periods of 4 months and 4 days at 4°C and 50°C, respectively. The K m and V max values of the purified xylose isomerase from G. thermodenitrificans TH2 were calculated as 32 mM and 4.68 μmol/min per mg of protein, respectively. Additionally, it was detected that some metal ions affected the enzyme activity at different ratios. The enzyme was active and stable at high temperatures and nearly neutral pHs which are desirable for the usage in the food and ethanol industry.

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Authors and Affiliations

  1. Department of Chemistry, Karadeniz Technical University, 61080, Trabzon, Turkey

    L. Konak, Y. Kolcuoğlu, E. Ozbek, A. Colak & B. Ergenoglu

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  1. L. Konak
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  2. Y. Kolcuoğlu
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  3. E. Ozbek
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  4. A. Colak
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  5. B. Ergenoglu
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Correspondence to A. Colak.

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Published in Russian in Prikladnaya Biokhimiya i Mikrobiologiya, 2014, Vol. 50, No. 1, pp. 34–38.

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Konak, L., Kolcuoğlu, Y., Ozbek, E. et al. Purification and characterization of an extremely stable glucose isomerase from Geobacillus thermodenitrificans TH2. Appl Biochem Microbiol 50, 25–29 (2014). https://doi.org/10.1134/S0003683814010062

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  • DOI: https://doi.org/10.1134/S0003683814010062

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