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Biocatalytic properties of thermolysin immobilized on polyvinyl alcohol cryogel

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Abstract

Preparations with different contents of thermolysin were obtained by the immobilization of the enzyme on granulated polyvinyl alcohol cryogel. Their activity and stability in an aqueous medium and in mixtures of polar organic solvents of different composition were investigated. The catalytic properties of the preparations in reactions of peptide bond formation were studied, and the optimal amount of the biocatalyst, the concentrations of initial reagents, and the ratios of organic solvents and water necessary for effective enzymatic peptide synthesis catalyzed by immobilized thermolysin were determined. A series of peptides of the general formula Z-Ala-Ala-Xaa-pNA, where Xaa = Leu, Ile, Phe, Val, or Ala, were synthesized, and the immobilized enzyme was shown to retain substrate specificity in an organic medium.

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Abbreviations

Dnp:

2,4-dinitrophenyl

pNA:

p-nitroanilide

PVA:

polyvinyl alcohol ThL, thermolysin

Z:

benzyloxycarbonyl

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Authors and Affiliations

  1. Nesmeyanov Institute of Organoelement Compounds, Russian Academy of Sciences, Moscow, Russia

    A. V. Belyaeva & V. I. Lozinskii

  2. Faculty of Bioengineering and Bioinformatics, Moscow State University, Vorob’evy Gory, Moscow, 119992, Russia

    Yu. A. Smirnova

  3. Faculty of Chemistry, Moscow State University, Vorob’evy gory, Moscow, 119992, Russia

    E. N. Lysogorskaya, E. S. Oksenoit & I. Yu. Filippova

  4. Belozersky Research Institute of Physicochemical Biology, Moscow State University, Vorob’evy gory, Moscow, 119992, Russia

    A. V. Timofeeva

Authors
  1. A. V. Belyaeva
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  2. Yu. A. Smirnova
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  3. E. N. Lysogorskaya
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  4. E. S. Oksenoit
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  5. A. V. Timofeeva
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  6. V. I. Lozinskii
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  7. I. Yu. Filippova
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Corresponding author

Correspondence to I. Yu. Filippova.

Additional information

Original Russian Text © A.V. Belyaeva, Yu.A. Smirnova, E.N. Lysogorskaya, E.S. Oksenoit, A.V. Timofeeva, V.I. Lozinskii, I.Yu. Filippova, 2008, published in Bioorganicheskaya Khimiya, 2008, Vol. 34, No. 4, pp. 487–494.

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Belyaeva, A.V., Smirnova, Y.A., Lysogorskaya, E.N. et al. Biocatalytic properties of thermolysin immobilized on polyvinyl alcohol cryogel. Russ J Bioorg Chem 34, 435–441 (2008). https://doi.org/10.1134/S1068162008040079

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  • DOI: https://doi.org/10.1134/S1068162008040079

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