Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
Josten, A; Meusel, M; Spener, F; Haalck, L.
Enzyme immobilization via microbial transglutaminase: a method for the generation of stable sensing surfaces
J MOL CATAL B-ENZYM. JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC; 7: 57-66.
Doi: 10.1016/S1381-1177(99)00021-1
Web of Science
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- Co-Autor*innen der Med Uni Graz
- Spener Friedrich
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- Abstract:
- Transglutaminase from a variant of Streptoverticillium mobaraense (MTGase), which catalyzes the acyl-transfer between gamma-carboxyamide groups and various primary amines was applied for the immobilization of biomolecules on electrode surfaces. Upon addition of suitable polypeptides (poly-L-lysine, poly-L-glutamine) or other proteins, which serve as substrates for MTGase, such as casein, gelatin or fibrinogen, networks can be generated by the entrapped and/or covalently attached sensing enzymes. These matrix proteins were used as cross-linkers and allow for the preparation of an enzyme layer with tailor made properties. The preparation of MTGase catalyzed enzyme layers on disposable screen printed electrodes using glucose oxidase (GOx) as a model enzyme is described. Best results were;achieved using 2.3 U GOx mm(-2) and 0.1 mg (poly-L-lysine/poly-L-glutamine) mm(-2), giving rise to a sensitivity of 0.15 nA 1 mu mol(-1) mm(-2). In a further approach the storage as well as the operational stability of lactate enzyme electrodes obtained upon immobilization of lactate oxidase (LOx) in a fibrinogen network was determined in comparison to biosensors prepared by classical chemical cross-linking. After storage of 20 weeks at 4 degrees C in humid atmosphere, the MTGase based electrodes remained fully stable as judged by their sensitivity, whereas the electrodes obtained via glutaraldehyde (GDA) cross-linking lost 40% of their sensitivity. The lifetime (LT50) of the MTGase derived electrodes was determined at ambient temperature to be 133 +/- 26 h tit = 4), whereas a lifetime of only 53 +/- 9 h (n = 3) was found for the electrodes with GDA cross-linking. (C) 1999 Elsevier Science B.V. All rights reserved.
- Find related publications in this database (Keywords)
- microbial transglutaminase
- immobilization
- stabilization
- protein cross-linking
- fibrinogen