Stabilization of proteases by entrapment in a new composite hydrogel |
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| Authors: | E A Markvicheva N E Tkachuk S V Kuptsova T N Dugina S M Strukova Yu E Kirssh V P Zubov L D Rumish |
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| Institution: | (1) Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, 117871 Moscow, Russia;(2) Moscow State University, Moscow, Russia;(3) Karpou Institute of Physical Chemistry, Moscow, Russia |
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| Abstract: | A new one-step procedure for entrapping proteases into a polymeric composite calcium alginate-poly(N-vinyl caproladam) hydrogel was developed that provided 75–90% retention of the activity of entrapped enzymes compared to
soluble ones. Properties of entrapped carboxypeptidase B, trypsin, and thrombin were investigated. The immobilized enzymes
were active within a wide pH range. The temperature optima of entrapped trypsin and carboxypeptidase B were approx 25°C higher
than that of the soluble enzymes, and the resistance to heating was also increased. The effects of various polar and nonpolar
organic solvents on the entrapped proteases were investigated. The immobilized enzymes retained their activity within a wide
concentration range (up to 90%) of organic solvents. Gel-entrapped trypsin and carboxypeptidase (CPB) were successfully used
for obtaining human insulin from recombinant proinsulin. The developed stabilization method can be used to catalyze various
reactions proceeding within wide pH and temperature ranges. |
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| Keywords: | Poly(N-vinyl caproladam) hydrogel thrombin trypsin carboxypeptidase B enzyme entrapment stabilization of proteases |
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