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Effect of salts and Triton X-100 on ultrafiltration purification and properties of extracellular glucose oxidase from Penicillium adametzii LF F-2044.1

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Abstract

We compared the effectiveness of glucose oxidase isolation from the culture fluid of Penicillium adametzii LF F-2044.1 in the presence of ammonium sulfate, ammonium chloride, and Triton X-100. Ammonium chloride inhibited glucose oxidase in the culture fluid. This compound increased K M (by 1.2–1.3 times), but decreased V max for D-glucose oxidation (by 1.7–1.8 times). Ammonium sulfate had little effect on kinetic parameters. Combined treatment with salts and Triton X-100 was followed by a significant increase in the effectiveness of ultrafiltration purification of the culture fluid. The samples of glucose oxidase were electrophoretically characterized. The dependence of kinetic parameters on glucose oxidase concentration during oxidation of D-glucose was evaluated. The catalytic constant and k cat/K M ratio for glucose oxidase samples from the culture fluid isolated in the presence of additives significantly surpassed those for enzyme samples, which were obtained by ultrafiltration of the culture fluid with no additives and chromatography on aluminum oxide. The activity of glucose oxidase isolated from the culture fluid in the presence of ammonium chloride was lower compared to that of the enzyme obtained in the presence of ammonium sulfate. This agent is preferable for ultrafiltration of the culture fluid.

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Abbreviations

GO:

glucose oxidase

CD:

circular dichroism

CF:

culture fluid filtrate

SAS:

surface-active substance

UFM:

ultrafiltration membrane

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Authors and Affiliations

  1. Institute of Bioorganic Chemistry, National Academy of Sciences of Belarus, Minsk, 220141, Belarus

    A. N. Eryomin

  2. Institute of Microbiology, National Academy of Sciences of Belarus, Minsk, 220141, Belarus

    L. A. Zhukovskaya & R. V. Mikhailova

Authors
  1. A. N. Eryomin
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  2. L. A. Zhukovskaya
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  3. R. V. Mikhailova
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Correspondence to A. N. Eryomin.

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Original Russian Text © A.N. Eryomin, L.A. Zhukovskaya, R.V. Mikhailova, 2009, published in Prikladnaya Biokhimiya i Mikrobiologiya, 2009, Vol. 45, No. 3, pp. 277–286.

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Eryomin, A.N., Zhukovskaya, L.A. & Mikhailova, R.V. Effect of salts and Triton X-100 on ultrafiltration purification and properties of extracellular glucose oxidase from Penicillium adametzii LF F-2044.1. Appl Biochem Microbiol 45, 248–257 (2009). https://doi.org/10.1134/S0003683809030028

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  • DOI: https://doi.org/10.1134/S0003683809030028

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