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Purification and biochemical characterization of a novel secretory dipeptidyl peptidase IV from porcine serum

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Abstract

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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Abbreviations

DPP-IV:

Dipeptidyl peptidase IV

GLP-1:

Glucagon-like peptide 1

Gly-Pro-AMC:

Glycyl prolyl 7-amino 4-methyl coumarin

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Acknowledgements

The authors thank Indian Council of Medical Research, New Delhi, India, for the financial assistance through Senior Research Fellowship granted to DK (ICMR Approval Letter No: 45/33/2012/BMS/TRM) to conduct this study. SNS acknowledges the financial support from the Department of Science and Technology, New Delhi, Government of India, under the project vide No DST/INDO/SOUTH AFRICA/P-12/2014.

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Authors and Affiliations

  1. Department of Biotechnology, Sri Jayachamarajendra College of Engineering, JSS Science and Technology University, JSS TI Campus, Mysuru, Karnataka, 570006, India

    Divya Kumar, K. N. Neema & Nanjunda Swamy Shivananju

  2. Department of Studies in Chemistry, University of Mysore, Manasagangothri, Mysuru, Karnataka, 570006, India

    Priya Babu Shubha

  3. Faculty of Natural Sciences, Adichunchanagiri University, Bellur Cross, B.G. Nagara, Mandya, Karnataka, India

    Vivek K. Hamse

  4. Department of Biotechnology, NMAM Institute of Technology, Nitte, Karkala Taluk, Udupi, Karnataka, 574110, India

    D. M. Chetan

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  1. Divya Kumar
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Correspondence to Nanjunda Swamy Shivananju.

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Kumar, D., Hamse, V.K., Neema, K.N. et al. Purification and biochemical characterization of a novel secretory dipeptidyl peptidase IV from porcine serum. Mol Cell Biochem 471, 71–80 (2020). https://doi.org/10.1007/s11010-020-03766-y

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