Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

    Full text documents are added when available. The database is updated daily and currently holds about 240,000 items, of which 72,000 in open access.

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Record number 360377
Title Reconstitution of apoglucose oxidase with FAD conjugates for biosensoring of progesterone
Author(s) Posthuma-Trumpie, G.A.; Berg, W.A.M. van den; Wiel, D.F.M. van de; Schaaper, W.M.M.; Korf, J.; Berkel, W.J.H. van
Source Biochimica et Biophysica Acta. Proteins & Proteomics 1774 (2007)7. - ISSN 1570-9639 - p. 803 - 812.
DOI https://doi.org/10.1016/j.bbapap.2007.04.009
Department(s) Livestock Research
Biochemistry
Central Institute for Animal Disease Control
VLAG
Publication type Refereed Article in a scientific journal
Publication year 2007
Keyword(s) flavin-adenine-dinucleotide - apoenzyme reactivation immunoassay - glucose-oxidase - aspergillus-niger - colorimetric immunoassays - enzyme-electrode - binding - label - milk - intermediate
Abstract The reconstitution of Aspergillus niger apoglucose oxidase (apoGOx) with FAD conjugates for biosensoring of progesterone was investigated. ApoGOx prepared by partial unfolding of the protein under acidic conditions consisted of reconstitutable monomers (50 ± 10%), reconstitutable dimers (20 ± 10%) and irreversibly aggregated oligomers (30 ± 20%). Incubation of monomeric apoGOx with FAD or N6-(6-aminohexyl)-FAD (ahFAD) restored glucose oxidase (GOx) activity and induced dimerization with stoichiometric incorporation of FAD. N6-(6-aminohexyl)-FAD progesterone conjugates also induced dimerization. However, holoenzyme reconstitution required relatively high concentrations of apoprotein and was dependent on the type of conjugate. Restoration to 25¿50% of the original enzyme activity was obtained. Binding of the FAD-progesterone conjugates might hinder the closure of a protein lid needed for dimer formation. Our results illustrate the prospects of FAD conjugates in sensitive detection of progesterone in biological matrices in a biosensor based on the recombination of apoGOx with progesterone-conjugated FAD.
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