Purification and biochemical characterization of a novel secretory dipeptidyl peptidase IV from porcine serum.
Mol Cell Biochem
; 471(1-2): 71-80, 2020 Aug.
Article
em En
| MEDLINE
| ID: mdl-32577945
Purification of DPP-IV enzyme from porcine serum, is presented in this study for the first time. The high molecular weight DPP-IV from porcine serum was fractioned using Sephadex G-75 gel filtration followed by DEAE Sephadex anion exchange and Sephadex G-100 gel filtration chromatography columns with a final yield of 11.25%. The SDS-PAGE of the purified sample showed a single band of molecular mass nearing 160 kDa. Distinct single band was observed after PAS staining confirmed it to be a glycoprotein. The purified enzyme showed an optimum pH and temperature of 8 and 37 °C, respectively. The enzyme effectively cleaved fluorogenic substrate Gly-Pro-AMC with Km and Vmax of 4.578 µM and 90.84 nmoles/min, respectively. Purified DPP-IV activity was inhibited by Diprotin A with an IC50 value of 8.473 µM. Among the three plant extracts used to study DPP-IV inhibition, the aqueous hot extract of Terminalia chebula showed the highest inhibition of 87.19%, followed by the aqueous cold extract of Momordica carantia, ( 31.6%) and Azadirachta indica (34.16%) at the concentration of 25 µg.
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Base de dados:
MEDLINE
Assunto principal:
Oligopeptídeos
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Dipeptidil Peptidase 4
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Dipeptídeos
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Ensaios Enzimáticos
Idioma:
En
Revista:
Mol Cell Biochem
Ano de publicação:
2020
Tipo de documento:
Article
País de afiliação:
Índia