Bromelain has a wide range of therapeutic efficacy: it demonstrates in vitro and in vivo anti-inflammatory, antithrombotic and fibrinolytic activities. The enzyme modulates the functions of adhesion molecules in the blood and endothelial cells, as well as regulates and activates various immune cells and cytokine production. The enzyme is used as an alternative or additional preparation for glucocorticoids, non-steroidal antirheumatic preparations and immunomodulating agents. In the present work, the physico-chemical and kinetic properties of bromelain in free and immobilized on the chitosan matrix states were studied. It has been established that the temperature range for the functioning of soluble and sorbed bromelain ranges from 50 to 70ºC; the optimum pH for the functioning of the native and immobilized enzyme is 7.5; the maximum rate of enzymatic catalysis of free and immobilized preparations is observed at azocasein concentrations of 0.4 mM; the maximum reaction rate for bromelain immobilized on medium molecular chitosan decreases 2.2 times, and for enzyme sorbed on high molecular chitosan it decreases 1.7 times. Immobilization of bromelain on the chitosan matrix does not lead to a shift in the temperature optimum, but increases the thermal stability of the enzyme.
bromelain, immobilization, physico-chemical properties of enzymes, chitosan
1. Amini A., Masoumi-Moghaddam S., Morris D.L. Utility of bromelain and N-acetylcysteine in treatment of peritoneal dissemination of gastrointestinal mucin-producing malignancies. Springer, 2016, 246 p.
2. Homaei A.A., Sajedi R.H., Sariri R., Seyfzadeh S., Stevanato R. Cysteine enhances activity and stability of immobilized papain. Amino Acids, 2010, vol. 38, pp. 937-942. DOI:https://doi.org/10.1007/s00726-009-0302-3
3. Martins B.C., Rescolino R., Coelho D.F., Espindola F.S., Zanchetta B., Tambourgi E.B., Silveira E. Studies of stability and characterization this enzyme bromelain in pineapple (Ananas comosus). BMC Proceedings, 2014, vol. 8, p. 137. DOI:https://doi.org/10.1186/1753-6561-8-S4-P137
4. Glik B., Pasternak D. Molekulyarnaya biotehnologiya. Principy i primenenie. M.: Mir, 2002, 589 s. @@[Glik B., Pasternak D. Molecular biotechnology. Principles and application. M.: Mir, 2002, 589 p. (In Russ.)]
5. Keleti T. Osnovy fermentativnoy kinetiki. M.: Mir, 1990, 350 s. @@[Keleti T. Basics of enzymatic kinetics. M.: Mir, 1990, 350 p. (In Russ.)]
6. Lenindzher A. Osnovy biohimii. T. 1. M.: Mir, 1985, 365 s. @@[Lenindzher A. Basics of biochemistry. Vol. 1. M.: Mir, 1985, 365 p. (In Russ.)]
7. Husain S.S., Lowe G. The amino acid sequence around the active-site cysteine and histidine residues, and the buried cysteine residue in ficin. Biochem. J, 1970, vol. 117, pp. 333-340.
8. Devaraj K.B., Kumar P.R., Prakash V. Purification, characterization, and solvent-induced thermal stabilization of ficin from Ficus carica. J. Agric. Food Chem., 2008, vol. 56, pp. 11417-11423. DOI:https://doi.org/10.1021/jf802205a.
9. Skryabin K.G. Hitin i hitozan. Poluchenie, svoystva i primenenie. M.: Nauka, 2002, 368 s. @@[Skryabin K.G. Chitin and chitosan. Preparation, properties and application. M.: Nauka, 2002, 368 p. (In Russ.)]
10. Magnin D., Lefebvre J., Chornet E., Dumitriu S. Physicochemical and structural characterization of a polyionic matrix of interest in biotechnology, in the pharmaceutical and biomedical fields. Carbohydrate Polymers, 2004, vol. 55, pp. 437-453. DOI:https://doi.org/10.1016/j.carbpol.2003.11.013.
11. Kumar M.N.V.R., Muzzarelli R.A.A., Muzzarelli C., Sashiwa H., Domb A.J., Chitosan chemistry and pharmaceutical perspectives. Chem. Rev., 2004, vol. 104, pp. 6017-6084. DOI:https://doi.org/10.4236/ajac.2014.510077.
12. Holyavka M.G., Artyuhov V.G., Koroleva V.A. Sposob polucheniya geterogennogo preparata razlichnoy dispersnosti na osnove bromelayna i hitozana. Patent na izobretenie № 2677232, 2019. @@[Holyavka M.G., Artyuhov V.G., Koroleva V.A. The method of obtaining a heterogeneous preparation of different dispersion based on bromelain and chitosan. Patent for invention № 2677232, 2019. (In Russ.)]
13. Koroleva V.A., Holyavka M.G., Sazykina S.M., Ol'shannikova S.S., Artyuhov V.G. Issledovanie sorbcionnoy emkosti kislotorastvorimyh hitozanov po otnosheniyu k bych'emu syvorotochnomu al'buminu. Vestnik VGU. Seriya «Himiya. Biologiya. Farmaciya», 2015, № 4, s. 85-89. @@[Koroleva V.A., Holyavka M.G., Sazykina S.M, Olshannikova S.S., Artyukhov V.G. Study of the sorption capacity of acid-soluble chitosans toward bovine serum albumin. Vestnik VGU. Seriya «Himiya. Biologiya. Farmaciya», 2015, no. 4, pp. 85-89. (In Russ.)]
