Biomedical Chemistry: Research and Methods 2024, 7(3), e00234

Immobilization of L-Asparaginase on Oxidized Bacterial Cellulose to Improve the Thermal Stability of the Enzyme

A.N. Shishparenok1*, S.A. Koroleva2, I.D. Zlotnikov3, Yu.A. Gladilina1, M.V. Pokrovskaya1, S.S. Alexandrova1, D.D. Zhdanov1

Institute of Biomedical Chemistry, 10 Pogodinskaya str., Moscow, 119121 Russia; *e-mail: a.shishparyonok@yandex.ru

Keywords: L-asparaginase; bacterial cellulose; immobilization; thermal stability

DOI:10.18097/BMCRM00234

The whole version of this paper is available in Russian.

Bacterial cellulose (BC) membranes can be modified for covalent immobilization of macromolecules. One type of modification is oxidation, after which the oxidized BC membrane (OBC) could be used as a matrix for covalent immobilization of enzymes. In this work, the BC membrane was chemically oxidized with sodium periodate (NaIO4) to increase the stability of immobilized mesophilic L-asparaginase (L-ASNase) from Erwinia carotovora (EwA). IR spectroscopy confirmed the immobilization of L-ASNase EwA on OBC membranes. Immobilization of the enzyme increased its temperature optimum for its activity by 15°C and raised the inactivation temperature to 60°C. The OBC membrane could be used as a potential carrier for covalent immobilization of enzymes to improve their pharmacological properties by increasing their thermostability.
Figure 1. IR spectra of BC, OBC, and OBC films with immobilized L-ASNase EwA. (A) Spectral range 4000-1000 cm-1. (B) Spectral range 2000-1000 cm-1
Figure 2. Temperature dependence of L-ASNase EwA enzyme activity. The enzyme was immobilized by physical adsorption on BC or covalently on OBC. The membrane with the enzyme was placed in buffer with substrate and incubated in the temperature range of 45-60°C. Enzyme activity was shown on membranes incubated at (A) 45°C, (B) 50°C, (C) 55°C, or (D) 60°C.

CLOSE
Table 1. Physicochemical characteristics of BC and OBC.

FUNDING

The work was conducted in accordance with framework of the Russian Federation fundamental research program for the long-term period (2021-2030) (No. 122022800499-5).

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