Specific immobilization of laccase onp-benzoquinone-activated supports |
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Authors: | Mircea-Alexandru Mateescu Enzo Agostinelli Grazyna Weltrowska Marek Weltrowski Bruno Mondovi |
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Affiliation: | (1) Département de chimie et GREFA, Universit'e du Québec à Montréal, C.P. 8888, succursale A, H3C 3P8 Montr'eal, Québec, Canada;(2) Départment of Biochemical Sciences and CNR - Center of Molecular Biology, Rome University La Sapienza, I-00185 Rome, Italy |
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Abstract: | Summary A specific immobilization of laccase (EC 1.10.3.2) onto a ready-to-usep-benzoquinone-activated agarose support is described. The single-step procedure leads to a laccase protein coupling of I8% and an enzyme activity immobilization yield of 27%, while the retained specific activity of the immobilized enzyme was 150% of the specific activity of the free laccase. This peculiar result is thought to be related to the fact that during the process of support activation byp-benzoquinone, a significant amount of the hydroquinone by-product of the activation process is coupled to the support. These coupled derivatives constitute substrate (hydroquinone) analogues for which laccase exhibits a high affinity. Therefore, simultaneous affinity retention on the hydroquinone groups and covalent coupling on the p-benzoquinone groups allow the binding of the enzyme in an advantageous conformation which can generate this increase specific activity by immobilization. The entire process can be considered as an affinity immobilization. The immobilized enzyme is much more stable to the inhibitory action of chloride and azide ions, with a recovery of 100% of the activity, than the free laccase, with a recovery of 67% and 32%, respectively, after removal of the inhibitors by dialysis. The stability was 95% after storage for 14 months at 4° C.Abbreviations HQ hydroquinone - p-BQ p-benzoquinone - U enzyme unitsPart of the work was presented at the Satellite FEBS 1989 Symposium onBiochemical and biophysical approaches to the study of copper proteins, Camerino, Italy. |
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Keywords: | Laccase Immobilization Agarose p-Benzoquinone Activated supports |
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