Modulating Substrate Specificity of Rhizobium sp. Histamine Dehydrogenase through Protein Engineering for Food Quality Applications
Yazarlar (7)
Karen Rodríguez-Núñez Universidad De La Serena
Alejandra Cortés-Monroy Universidad De La Serena
Marcela Serey Universidad De La Serena
Dr. Öğr. Üyesi Yunus ENSARİ Kafkas Üniversitesi, Türkiye
Mehdi D Davari Leibniz Institut Fur Pflanzenbiochemie
Claudia Bernal Universidad De La Serena
Ronny Martinez Universidad De La Serena
| Makale Türü |
Özgün Makale
(SSCI, AHCI, SCI, SCI-Exp dergilerinde yayınlanan tam makale)
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| Dergi Adı | MOLECULES (Q2) | ||
| Dergi ISSN | 1420-3049 Wos Dergi Scopus Dergi | ||
| Dergi Tarandığı Indeksler | SCI | ||
| Makale Dili | Türkçe | Basım Tarihi | 04-2023 |
| Kabul Tarihi | 12-04-2026 | Yayınlanma Tarihi | – |
| Cilt / Sayı / Sayfa | 28 / 9 / 3748–0 | DOI | 10.3390/molecules28093748 |
| Makale Linki | http://dx.doi.org/10.3390/molecules28093748 | ||
| Özet |
| Histamine is a biogenic amine found in fish-derived and fermented food products with physiological relevance since its concentration is proportional to food spoilage and health risk for sensitive consumers. There are various analytical methods for histamine quantification from food samples; however, a simple and quick enzymatic detection and quantification method is highly desirable. Histamine dehydrogenase (HDH) is a candidate for enzymatic histamine detection; however, other biogenic amines can change its activity or produce false positive results with an observed substrate inhibition at higher concentrations. In this work, we studied the effect of site saturation mutagenesis in Rhizobium sp. Histamine Dehydrogenase (Rsp HDH) in nine amino acid positions selected through structural alignment analysis, substrate docking, and proximity to the proposed histamine-binding site. The resulting libraries were screened for histamine and agmatine activity. Variants from two libraries (positions 72 and 110) showed improved histamine/agmatine activity ratio, decreased substrate inhibition, and maintained thermal resistance. In addition, activity characterization of the identified Phe72Thr and Asn110Val HDH variants showed a clear substrate inhibition curve for histamine and modified kinetic parameters. The observed maximum velocity (Vmax) increased for variant Phe72Thr at the cost of an increased value for the Michaelis–Menten constant (Km) for histamine. The increased Km value, decreased substrate inhibition, and biogenic amine interference observed for variant Phe72Thr support a tradeoff between substrate affinity and substrate inhibition … |
| Anahtar Kelimeler |
| agmatine | dehydrogenase | diaminopropane | enzyme engineering | histamine | protein design |
BM Sürdürülebilir Kalkınma Amaçları
| Atıf Sayıları | |
| Google Scholar | 8 |
| Web of Science | 7 |
| Scopus | 7 |