Immobilization of trypsin on chitosan gels: use of different activation protocols and comparison with other supports.
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.
Manrich
Anny
A
Departamento de Engenharia Química, Universidade Federal de São Carlos, Rodovia Washington Luiz, São Carlos, SP, Brazil.
Galvão
Célia M A
CM
Jesus
Charles D F
CD
Giordano
Roberto C
RC
Giordano
Raquel L C
RL
D000143:Acids / Q000737:chemistry; D000468:Alkalies / Q000737:chemistry; D000818:Animals; D002417:Cattle; D048271:Chitosan / Q000737:chemistry*; D004789:Enzyme Activation; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry*; D005782:Gels; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D010179:Pancreas / Q000201:enzymology; D012995:Solubility; D013696:Temperature; D014357:Trypsin / Q000737:chemistry* / Q000378:metabolism*
D000143:Acids; D000468:Alkalies; D004800:Enzymes, Immobilized; D005782:Gels; D048271:Chitosan; D014357:Trypsin