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Immobilization of proteases in composite hydrogel based on poly(N-vinyl caprolactam)

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Biotechnology Techniques

Summary

A novel one-step method was developed for entrapment of proteolytic enzymes, namely carboxypeptidase B and trypsin, in polymer composite gel based on poly(N-vinyl caprolactam). Native proteases and enzymes previously stabilized by covalent attachment to poly(vinylpyrrolidone-co-acrolein) were immobilized in gels. After immobilization, about 90% and 75% of original trypsin and CPB activities, respectively, were retained. The immobilized enzymes were active within a wide pH range. The optimum temperature of the entrapped enzymes was approximately 25°C higher than that of the soluble enzymes. The entrapped enzymes were successfully used to obtain human insulin from recombinant proinsulin.

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Authors and Affiliations

  1. Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya, 16/10, 117871, Moscow, Russia

    Markvicheva E. A., Bronin A. S., Kudryavtseva N. E., Rumsh L. D., Kirsh Yu. E. & Zubov V. P.

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  1. Markvicheva E. A.
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  2. Bronin A. S.
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  3. Kudryavtseva N. E.
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  4. Rumsh L. D.
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  5. Kirsh Yu. E.
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  6. Zubov V. P.
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Markvicheva, E.A., Bronin, A.S., Kudryavtseva, N.E. et al. Immobilization of proteases in composite hydrogel based on poly(N-vinyl caprolactam). Biotechnol Tech 8, 143–148 (1994). https://doi.org/10.1007/BF00161578

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  • DOI: https://doi.org/10.1007/BF00161578

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