Prevention of interfacial inactivation of enzymes by coating the enzyme surface with dextran-aldehyde.
Interactions between soluble enzymes and interfaces of organic solvent drops or gas bubbles have a very negative effect on the operational stability of the soluble enzymes. In this study, the formation of a hydrophilic shell around the enzyme has been attempted using dextran-aldehyde which would prevent the interaction between enzyme and hydrophobic interfaces with minimal modification of the enzyme surface. After optimizing the size of the dextran (that was found to play a critical role), three different enzymes (glucose oxidase, d-amino acid oxidase, and trypsin) have been conjugated with dextran-aldehyde and their stability towards organic-aqueous and air-liquid interfaces has been evaluated. The treatment itself proved to be very low-cost in terms of activity and was highly stabilizing for the three enzymes assayed. The conjugated preparation of the three assayed enzymes remained fully active in the presence of air-liquid interfaces for at least 10h. However, the unmodified enzymes lost more than 50% of activity within the first hour of the experiments except for trypsin which kept 38% activity after 12h while the trypsin dextran-aldehyde conjugate maintained 100% enzyme activity. Similar results were achieved in the presence of stirred organic solvent-aqueous buffer biphasic system, although in this case some activity was lost by the action of the soluble portion of the organic solvent. In fact, this treatment seems to be also effective to improve the resistance to the action of organic solvent.
Betancor
Lorena
L
Departamento de Biocatálisis, Instituto de Catálisis, CSIC, Campus Universidad Autónoma, Cantoblanco, 28049 Madrid, Spain.
López-Gallego
Fernando
F
Hidalgo
Aurelio
A
Alonso-Morales
Noelia
N
Fuentes
Manuel
M
Fernández-Lafuente
Roberto
R
Guisán
José M
JM
D000447:Aldehydes / Q000737:chemistry; D000818:Animals; D003605:D-Amino-Acid Oxidase / Q000378:metabolism*; D003911:Dextrans / Q000737:chemistry; D004789:Enzyme Activation; D004795:Enzyme Stability; D005949:Glucose Oxidase / Q000378:metabolism*; D011485:Protein Binding; D012995:Solubility; D013499:Surface Properties; D014357:Trypsin / Q000378:metabolism*
D000447:Aldehydes; D003911:Dextrans; D005949:Glucose Oxidase; D003605:D-Amino-Acid Oxidase; D014357:Trypsin