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Khounmirzaie, N., Jalali, M.R., Shahriari, A., Tabandeh, M.R.. (1400). Comparison of glutaraldehyde cross linking versus direct Schiff base reaction for conjugation of L-asparaginase to nano-chitosan and improvement of enzyme physicochemical properties. سامانه مدیریت نشریات علمی, 20(6), 1804-1821. doi: 10.22092/ijfs.2021.125502
N. Khounmirzaie; M.R. Jalali; A. Shahriari; M.R. Tabandeh. "Comparison of glutaraldehyde cross linking versus direct Schiff base reaction for conjugation of L-asparaginase to nano-chitosan and improvement of enzyme physicochemical properties". سامانه مدیریت نشریات علمی, 20, 6, 1400, 1804-1821. doi: 10.22092/ijfs.2021.125502
Khounmirzaie, N., Jalali, M.R., Shahriari, A., Tabandeh, M.R.. (1400). 'Comparison of glutaraldehyde cross linking versus direct Schiff base reaction for conjugation of L-asparaginase to nano-chitosan and improvement of enzyme physicochemical properties', سامانه مدیریت نشریات علمی, 20(6), pp. 1804-1821. doi: 10.22092/ijfs.2021.125502
Khounmirzaie, N., Jalali, M.R., Shahriari, A., Tabandeh, M.R.. Comparison of glutaraldehyde cross linking versus direct Schiff base reaction for conjugation of L-asparaginase to nano-chitosan and improvement of enzyme physicochemical properties. سامانه مدیریت نشریات علمی, 1400; 20(6): 1804-1821. doi: 10.22092/ijfs.2021.125502

Comparison of glutaraldehyde cross linking versus direct Schiff base reaction for conjugation of L-asparaginase to nano-chitosan and improvement of enzyme physicochemical properties

Iranian Journal of Fisheries Sciences
دوره 20، شماره 6، زمستان 2021، صفحه 1804-1821  XML اصل مقاله (1012.65 K)
شناسه دیجیتال (DOI): 10.22092/ijfs.2021.125502
نویسندگان
N. Khounmirzaie1؛ M.R. Jalali1؛ A. Shahriari2؛ M.R. Tabandeh2
1Department of Clinical Sciences, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran.
2Department of Basic Sciences, Division of Biochemistry and Molecular Biology, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran.
چکیده
The bacterial L-asparaginase (ASNase) has been used in the treatment of asparagine-associated tumors; however, the instability of the enzyme increases the number of injections as well as the side effects. In the present study, ASNase was conjugated to nanochitosan (ASNase -CSNPS) by direct (shiff-base) and indirect (glutaraldehyde linker) methods. In order to get the optimal conjugation, ASNase/CSNPS ratio was first investigated. The physicochemical properties (optimum pH, temperature, residual activity), enzyme kinetics (Michaelis constants; Km and maximal velocity; Vmax) and stability (against freezing, proteolysis, and chemical denaturation) were determined. The results showed that the highest residual enzyme activity (>85%) was obtained using a combination of ASNase and CSNPS at 1:5 mass ratio in both conjugation methods. ASNase -CSNPS prepared by glutaraldehyde linker had higher Km and Vmax values (69.7 μM, 20.6 mol/mL/minμ), wider range of optimum pH and higher temperature stability compared to ASNase-CSNPS produced by Schiff-base (Km: 105.8 μM, Vmax 14.5 mol/ml/minμ) method. ASNase-CSNPS produced by indirect method had more stability against freezing-thawing, and proteolysis when compared with ASNase-CSNPS prepared using direct method. The results showed that application of glutaraldehyde coupling was superior to Schiff base cross linking for conjugating of ASNase to CSNPS and for production of ASNase with better physicochemical properties for future cancer therapy.
کلیدواژه‌ها
L-Asparaginase؛ Nanochitosan؛ Glutaraldehyde linker؛ Schiff base reaction؛ physicochemical properties
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