Publication
Recombinant bovine serum albumin domain II as bioreceptor for ochratoxin A capture
| dc.contributor.author | Aguiar, Tatiana Q. | |
| dc.contributor.author | Leal, Tânia | |
| dc.contributor.author | Rodrigues, Diana G. | |
| dc.contributor.author | Abrunhosa, Luís | |
| dc.contributor.author | Oliveira, Carla | |
| dc.contributor.author | Domingues, Lucília | |
| dc.date.accessioned | 2024-11-08T14:35:29Z | |
| dc.date.available | 2024-11-08T14:35:29Z | |
| dc.date.issued | 2025-02-01 | |
| dc.description.abstract | Established chromatographic techniques for mycotoxin control in foodstuffs require prior sample enrichment and clean-up, typically achieved using immunoaffinity columns (IACs). Bovine serum albumin (BSA) has recently emerged as a cost-effective alternative to antibodies used in IACs. This study aimed at exploring the BSA domain II (BDII), which houses the primary binding site for ochratoxin A (OTA), as a bioreceptor for OTA capture. Recombinant BDII (rBDII) was produced in soluble form by Escherichia coli Origami 2(DE3), fused to a His6 (HisBDII) or thioredoxin-His6 (TrxBDII) tag, with yields up to 19 ± 4.3 mg/Lculture in shake-flask. Fluorescence and circular dichroism (CD) spectroscopy revealed interaction of OTA with both rBDII variants, with estimated binding constants for OTA-HisBDII/TrxBDII complexes in the range of 5.7–9.3 × 104 M−1. CD also showed an α/β structure of rBDII variants, in opposition to the predominant α-helical structure of whole BSA, and slight increase in their α-helical content upon binding to OTA. TrxBDII immobilized on Ni-NTA resin successfully captured OTA from spiked samples at the optimum pH range of 6.5–7.0, allowing OTA extraction, clean-up, and enrichment from spiked white grape juice, with up to 84 ± 7.4 % recovery. | pt_PT |
| dc.description.version | info:eu-repo/semantics/publishedVersion | pt_PT |
| dc.identifier.doi | 10.1016/j.talanta.2024.127126 | pt_PT |
| dc.identifier.eid | 85207880738 | |
| dc.identifier.issn | 0039-9140 | |
| dc.identifier.pmid | 39489069 | |
| dc.identifier.uri | http://hdl.handle.net/10400.14/47158 | |
| dc.identifier.wos | 001350335900001 | |
| dc.language.iso | eng | pt_PT |
| dc.peerreviewed | yes | pt_PT |
| dc.rights.uri | http://creativecommons.org/licenses/by/4.0/ | pt_PT |
| dc.subject | Bioreceptor | pt_PT |
| dc.subject | BSA domain II | pt_PT |
| dc.subject | Ochratoxin A | pt_PT |
| dc.subject | Recombinant protein | pt_PT |
| dc.subject | Solid-phase extraction | pt_PT |
| dc.title | Recombinant bovine serum albumin domain II as bioreceptor for ochratoxin A capture | pt_PT |
| dc.type | journal article | |
| dspace.entity.type | Publication | |
| oaire.citation.title | Talanta | pt_PT |
| oaire.citation.volume | 283 | pt_PT |
| rcaap.rights | openAccess | pt_PT |
| rcaap.type | article | pt_PT |