Bioaccessibility and In Vitro Intestinal Permeability of a Recombinant Lectin from Tepary Bean (Phaseolus acutifolius) Using the Everted Intestine Assay
| Autor: | Lineth J. Vega-Rojas, Konisgmar Escobar-García, Juan Mosqueda, Antonio Escobedo-Reyes, D.M. Palmerín-Carreño, Teresa García-Gasca, Alejandro Blanco-Labra, Ivan Luzardo-Ocampo |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
recombinant lectin Catalysis in vitro and ex vivo gastrointestinal digestion Inorganic Chemistry 03 medical and health sciences 0302 clinical medicine food In vivo medicine Physical and Theoretical Chemistry Molecular Biology Tepary Bean Spectroscopy Intestinal permeability biology Chemistry Organic Chemistry Tepary bean (Phaseolus acutifolius) Lectin General Medicine medicine.disease bioaccessibility food.food Small intestine Computer Science Applications ex vivo everted intestine assay 030104 developmental biology medicine.anatomical_structure Biochemistry apparent permeability 030220 oncology & carcinogenesis biology.protein Phaseolus acutifolius Digestion Ex vivo |
| Zdroj: | International Journal of Molecular Sciences Volume 22 Issue 3 |
| ISSN: | 1422-0067 |
| DOI: | 10.3390/ijms22031049 |
| Popis: | Tepary bean (Phaseolus acutifolius) lectins exhibit differential in vitro and in vivo biological effects, but their gastrointestinal interactions and digestion have not yet been assessed. This work aimed to evaluate the changes of a recombinant Tepary bean lectin (rTBL-1) through an in vitro and ex vivo gastrointestinal process. A polyclonal antibody was developed to selectively detect rTBL-1 by Western blot (WB) and immunohistochemical analysis. Everted gut sac viability was confirmed until 60 min, where protein bioaccessibility, apparent permeability coefficient, and efflux ratio showed rTBL-1 partial digestion and absorption. Immunoblot assays suggested rTBL-1 internalization, since the lectin was detected in the digestible fraction. The immunohistochemical assay detected rTBL-1 presence at the apical side of the small intestine, potentially due to the interaction with the intestinal cell membrane. The in silico interactions between rTBL-1 and some saccharides or derivatives showed high binding affinity to sialic acid (&minus 6.70 kcal/mol) and N-acetylglucosamine (&minus 6.10 kcal/mol). The ultra-high-performance liquid chromatography&ndash electron spray ionization&ndash quantitative time-of-flight coupled to mass spectrometry (UHPLC&ndash ESI&ndash QTOF/MS) analysis showed rTBL-1 presence in the gastric content and the non-digestible fraction after intestinal simulation conditions. The results indicated that rTBL-1 partially resisted the digestive conditions and interacted with the intestinal membrane, whereas its digestion allowed the absorption or internalization of the protein or the derivative peptides. Further purification of digestion samples should be conducted to identify intact rTBL-1 protein and digested peptides to assess their physiological effects. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|