Characterization of Stability of the Young Porcine Intestinal Alkaline Phosphatase as a Candidate Exogenous Biocatalyst
Notice bibliographique
Résumé
Intestinal alkaline phosphatase (IAP) is a critical anti‐inflammatory mediator through possessing the ability to catalyze the hydrolytic dephosphorylation of endotoxin lipopolysaccharides (LPS) and other emblematic members of pathogen‐associated‐molecular patterns (PAMPs) such as ATP, thus preventing gut dysbiosis and disorders. Our previous work has demonstrated that early weaning decreases IAP affinity, indicating an increased susceptibility to bowel inflammation and infection in weanling piglets. To determine if the porcine IAP is suitable to serve as an efficacious exogenous feed enzyme biocatalyst, several key stability aspects of the AP need to be thoroughly characterized, including thermostability, i.e., resistance to heating typically at 70 – 80°C for 8 to 10 min during compound feed pelleting, gastric stability and resistance to exocrine pancreatic proteases' hydrolysis. This study was conducted to investigate the stability of the porcine IAP with jejunal samples collected from four 10‐day‐old Yorkshire piglets. The IAP activity was measured in the media containing 1.75 mM P ‐nitrophenyl phosphate at pH 7.4 for 30 min after exposing homogenized porcine jejunal tissue samples (19 – 27 μg) to various stability effectors. Stability of the IAP was assessed through inhibition kinetic analysis by the Eadie‐Hofstee model, including parameter endpoints (parameter estimates ± SE, n = 24) of TC 50 representing the temperature (°C) or time (h) at half the maximal inhibition of the IAP activity; and I max representing the relative maximal level (% of the control) of inhibition in the IAP activity. Thermostability of the porcine IAP was determined by heating the tissue homogenate at the temperature ranging from 22 – 80 °C for 10 min, and was associated with the inhibition kinetics of TC 50 = 24.5 ± 3.2 °C and I max = 80.0 ± 20.0 %. Gastric stability of the porcine IAP was determined by exposing the tissue homogenate to the pH at 3.5 for 0 – 5 h, and was associated with the inhibition kinetics of TC 50 = 0.40 ± 0.06 h and I max = 81.5 ± 3.2 %. Resistance of the porcine IAP to the gastric protease was determined by exposing the tissue homogenate to the 266 U/mL pepsin in the media for 0 – 5 h, and was associated with the inhibition kinetics of TC 50 = 0.46 ± 0.03 h and I max = 92.4 ± 0.8 %. Resistance of the IAP to the exocrine pancreatic proteases was determined by exposing the tissue homogenate to the 66 U/mL trypsin and 59 U/mL chymotrypsin in the media for 0 – 5 h, and was associated with the inhibition kinetics of TC 50 = 0.24 ± 0.11 h and I max = 11.7 ± 1.2 % for the trypsin effect; and of TC 50 = 0.09 ± 0.14 h and I max = 18.54 ± 0.95 % for the chymotrypsin effect, respectively. These results suggest that while the porcine IAP displays a high level of resistance to pancreatic proteases, it is not thermostable, and it is also very susceptible to the deactivation by the gastric acidic pH and pepsin environment. Strategies to improve the gastric acidic pH, protease and heat stability of the porcine IAP are required as a pre‐requisite to use this enzyme as an exogenous feed enzyme. Support or Funding Information NSERC Discovery Program of Canada and Metagen Enzyme Corporation
Récupéré en direct depuis OpenAlex et désinversé. Les résumés ne sont pas conservés dans cette base de données : les index inversés représentent 8,6 Go des 9,3 Go de texte de la base, et le serveur dispose de 13 Go libres.
Comment cette classification a été obtenuedéplier
Prédiction distillée sur la base complète
Imitation des enseignantsNi prévalence calibrée, ni vérité terrain. Validation humaine à venir. Apprise à partir de 10 348 étiquettes directes de Codex et de 10 348 étiquettes directes de Gemma. Le mode candidate est l'union des têtes enseignantes seuillées; le consensus est leur intersection. Ces sorties portent le statut machine_predicted_unvalidated et ne sont ni des étiquettes humaines ni des étiquettes directes de modèles de pointe.
Scores Codex et Gemma par catégorie
| Catégorie | Codex | Gemma |
|---|---|---|
| Métarecherche | 0,002 | 0,002 |
| Méta-épidémiologie (sens strict) | 0,000 | 0,000 |
| Méta-épidémiologie (sens large) | 0,000 | 0,000 |
| Bibliométrie | 0,000 | 0,000 |
| Études des sciences et des technologies | 0,001 | 0,001 |
| Communication savante | 0,000 | 0,000 |
| Science ouverte | 0,001 | 0,000 |
| Intégrité de la recherche | 0,000 | 0,000 |
| Charge utile insuffisante (le modèle a refusé de juger) | 0,000 | 0,000 |
Scores machine (provisoires)
Les deux têtes enseignantes du modèle étudiant, lues sur ce travail. Un score ordonne la base pour la relecture; il n'affirme jamais une catégorie, et le statut de validation accompagne chaque rangée tel quel.
Scores de référence d'un modèle non mature (critères de maturité non atteints, 7 itérations). Un score ordonne; il n'affirme jamais une catégorie.
score_only:v0-immature-baseline · tel quel depuis la passe de notation : score_only signifie que le nombre peut ordonner les travaux, et qu'aucune étiquette de catégorie n'en découleClassification
machine, non validéePrédiction automatique; un appel candidat d’une seule tête enseignante, pas un consensus.
Le détail, modèle par modèle et score par score, se trouve en fin de page sous « Comment cette classification a été obtenue ».