Immobilization of trypsin on chitosan gels: use of different activation protocols and comparison with other supports

International Journal of Biological Macromolecules
Anny ManrichR L C Giordano

Abstract

Trypsin was immobilized on chitosan gels coagulated with 0.1 or 1 M NaOH and activated with glutaraldehyde or glycidol. The derivatives were characterized by their recovered activity, thermal (40, 55 and 70 degrees C) and alkaline (pH 11) stabilities, amount of enzyme immobilized on gels for several enzyme loads (8-14 mg(protein)/g(Gel)) and compared to agarose derivatives. Enzyme loads higher than 14 mg(protein)/g(Gel) can be immobilized on glutaraldehyde derivatives, which showed 100% immobilization yield and, for loads up to 8 mg(protein)/g(Gel), 100% recovered activity. Activation with glycidol led to lower immobilization yields than the ones obtained with glutaraldehyde, 61% for agarose-glyoxyl (AgGly) with low grade of activation and 16% for the chitosan-glyoxyl (ChGly), but allowed obtaining the most stable derivative (ChGly), that was 660-fold more stable than the soluble enzyme at 55 and 70 degrees C-approximately threefold more stable than AgGly. The ChGly derivative presented also the highest stability during incubation at pH 11. Analyses of lysine residue contents in soluble and immobilized trypsin indicated formation of multipoint bonds between enzyme and support, for glyoxyl derivatives.

References

Mar 1, 1996·Biomaterials·A Jayakrishnan, S R Jameela
Apr 5, 2003·Biotechnology Progress·Paulo W TardioliJosé M Guisán

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Citations

Feb 2, 2010·Applied Biochemistry and Biotechnology·Anny ManrichRaquel Lima Camargo Giordano
Dec 3, 2011·Enzyme and Microbial Technology·Arezoo KhosraviIran Alemzadeh
Dec 3, 2011·Enzyme and Microbial Technology·C R SilvaR L C Giordano
Dec 4, 2012·International Journal of Biological Macromolecules·Jae-Min ParkYang-Hoon Kim
Feb 18, 2015·Colloids and Surfaces. B, Biointerfaces·Keziban Atacan, Mahmut Özacar
Apr 4, 2021·Marine Drugs·Diana R BaidamshinaAirat R Kayumov
Apr 18, 2021·International Journal of Biological Macromolecules·Yale Luck NunesJosé C S Dos Santos

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