| Yazarlar |
Karen Rodríguez-Núñez
Türkiye |
|
Alejandra Cortés-Monroy
Türkiye |
|
Marcela Serey
Türkiye |
Dr. Öğr. Üyesi Yunus ENSARİ
Kafkas Üniversitesi, Türkiye |
|
Mehdi D Davari
Türkiye |
|
Claudia Bernal
Türkiye |
|
Ronny Martinez
Türkiye |
| 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 in the catalytic mechanism of HDHs. Considering this tradeoff for future enzyme engineering of HDH could lead to breakthroughs in performance increases and understanding of this enzyme class. |
| Anahtar Kelimeler |
| enzyme engineering | histamine | dehydrogenase | agmatine | diaminopropane | protein design |
| Makale Türü | Özgün Makale |
| Makale Alt Türü | SSCI, AHCI, SCI, SCI-Exp dergilerinde yayımlanan tam makale |
| Dergi Adı | MOLECULES |
| Dergi ISSN | 1420-3049 |
| Dergi Tarandığı Indeksler | SCI |
| Dergi Grubu | Q2 |
| Makale Dili | Türkçe |
| Basım Tarihi | 04-2023 |
| Cilt No | 28 |
| Sayı | 9 |
| Doi Numarası | 10.3390/molecules28093748 |
| Makale Linki | http://dx.doi.org/10.3390/molecules28093748 |
| Atıf Sayıları | |
| WoS | 2 |
| SCOPUS | 2 |
| Google Scholar | 2 |
