Abstract and keywords
Abstract (English):
Papain (EC 3.4.22.2) is an endolytic cysteine protease derived from papaya latex (Carica papaya L.). Today, this enzyme is considered as an anti-inflammatory, anticoagulant and hemolytic agent, as well as a substance that accelerates tissue repair. Chitosan is classified as a universal sorbent that binds a wide range of substances of organic and inorganic origin. It has antimicrobial activity, biodegradability, non-toxicity and bioadhesive properties. The aim of our work is to study the combined effect of UV-radiation and temperature on the activity of free papain and papain immobilized on a matrix of medium and high molecular weight chitosan. Papain was chosen as the object of study, bovine serum albumin (BSA) and azocasein served as substrates for hydrolysis, medium- (Mr = 200 kDa, degree of deacetylation (DD) 82%) and high molecular weight (Mr = 350 kDa, DD = 94.85%) chitosan were used as carriers for immobilization. It has been shown that the enzyme becomes more resistant to changes in the temperature regime, as well as to UV irradiation after its immobilization. At 90°C, native papain is completely inactivated, while the sorbed enzyme on medium and high molecular weight chitosan retains 45 and 57% of its initial activity, respectively. After the sorption of papain on the polysaccharide matrix at doses from 151 to 6040 J/m2, the catalytic ability of the enzyme changed to a lesser extent than that of its soluble form. The proteolytic ability of the biocatalyst immobilized on high molecular weight chitosan after UV irradiation even increased by 13-27% of the initial level.

Keywords:
papain, immobilization, chitosan, UV-irradiation
Text
Text (PDF): Read Download
References

1. Holyavka M., Koroleva V., Olshannikova S., Artyukhov V., Faizullin D., Zakhartchenko N., Zuev Y., Kondratyev M., Zakharova E. Novel biotechnological formulations of cysteine proteases, immobilized on chitosan. Structure, stability and activity. International Journal of Biological Macromolecules, 2021, vol. 180, pp. 161-176.

2. Holyavka M.G., Nakvasina M.A., Artyuhov V.G. Praktikum po biotekhnologii: immobilizovannye biologicheskie ob"ekty v sisteme laboratornyh rabot. Uchebnoe posobie, Voronezh, 2017 (In Russ.).

3. Holyavka M.G., Artyuhov V.G. Immobilizovannye biologicheskie sistemy: biofizicheskie aspekty i prakticheskoe primenenie. Uchebnoe posobie, Voronezh, 2017 (In Russ.).

4. Baidamshina D.R., Trizna E.Y. et al. Anti-biofilm and wound-healing activity of chitosan-immobilized ficin. International Journal of Biological Macromolecules, 2020, vol. 164, pp. 4205-4217.

5. Fileti A.M.F., Fischer G.A., Tambourgi E.B. Neural modeling of bromelain extraction by reversed micelles. Brazilian Archives of Biology and Technology, 2010, vol. 53, pp. 455-463, doi:https://doi.org/10.1590/S1516-89132010000200026.

6. Grzonka Z., Kasprzykowski F., Wiczk W. Cysteine proteases. Industrial Enzymes, Springer, New York. J. Polaina, 2007, pp. 181-195.

7. Konno K., Hirayama C., Nakamura M., Tateishi K., Tamura Y., Hattori M., Kohno K. Papain protects papaya trees from herbivorous insects: role of cysteine proteases in latex. The Plant Journal, 2004, vol. 37, pp. 370-378, doi:https://doi.org/10.1046/j.1365-313X.2003.01968.x.

8. Khanna N., Panda P.C. The effect of papain on tenderization and functional properties of spending hen meat cuts. Indian Journal of Animal Research, 2007, vol. 41, pp. 55-58.

9. Baidamshina D.R., Trizna E.Y., Kayumov A.R., Koroleva V.A., Olshannikova S.S., Artyukhov V.G., Holyavka M.G., Bogachev M.I. Biochemical properties and anti-biofilm activity of chitosan-immobilized papain. Marine Drugs, 2021, vol. 19, p. 197.

10. Silva D.F., Rosa H., Carvalho A.F.A., Neto P.O. Immobilization of papain on chitin and chitosan and recycling of soluble enzyme for deflocculation of Saccharomyces cerevisiae from bioethanol distilleries. Enzyme Research, 2015, art. id 573721, doi:https://doi.org/10.1155/2015/573721.

11. Albuquerque P.B.S., de Oliveira W.F., dos Santos Silva P.M., dos Santos Correia M.T., Kennedy J.F., Coelho L.C.B.B. Epiphanies of well-known and newly discovered macromolecular carbohydrates - A review. International Journal of Biological Macromolecules, 2020, vol. 156, pp. 51-66, doi:https://doi.org/10.1016/j.ijbiomac.2020.04.046.

12. Koroleva V.A., Holyavka M.G., Artyuhov V.G., Sazykina S.M., Timoshilova A.A., Olypannikova S.S. Razrabotka metodiki immobilizacii ficina na matricah kislotorastvorimyh hitozanov. V sbornike: Organizaciya i regulyaciya fiziologo-biohimicheskih processov. Mezhregional'nyj sbornik nauchnyh rabot. Voronezhskij gosudarstvennyj agrarnyj universitet im. Imperatora Petra I. Voronezh, 2015, pp. 99-102 (In Russ.).

13. Sabirova A.R., Rudakova N.L., Balaban N.P., Ilyinskaya O.N., Demidyuk I.V., Kostrov S.V., Rudenskaya G.N., Sharipova M.R. A novel secreted metzincin metalloproteinase from Bacillus intermedius. FEBS Lett., 2010, vol. 584, no. 21, pp. 4419-4425, doi:https://doi.org/10.1016/j.febslet.2010.09.049.


Login or Create
* Forgot password?