Tropical Journal of Pharmaceutical Research

Original Research Article | OPEN ACCESS

expression and Purification of Soluble, Biologically Active Recombinant Dipeptidyl Peptidase 4 (DPP4/CD26/ADAbp) Comprising the Extracellular Domain in the Yeast, Pichia pastoris

Hyun Kang

Department of Medical Laboratory Science, College of Health Science, Dankook University, Cheonan-si, Chungnam, 330-714, Republic of Korea;

For correspondence:- Email: hyunbio@gmail.com Tel:+82415501452

Received: 6 April 2014 Accepted: 10 May 2014 Published: 26 June 2014

Citation: Kang H. expression and Purification of Soluble, Biologically Active Recombinant Dipeptidyl Peptidase 4 (DPP4/CD26/ADAbp) Comprising the Extracellular Domain in the Yeast, Pichia pastoris. Trop J Pharm Res 2014; 13(6):855-862 doi: 10.4314/tjpr.v13i6.5

© 2014 The authors.
This is an Open Access article that uses a funding model which does not charge readers or their institutions for access and distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0) and the Budapest Open Access Initiative (http://www.budapestopenaccessinitiative.org/read), which permit unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited..

Abstract

Purpose: To investigate Pichia pastoris expression system for producing clinically usable, high-quality dipeptidyl peptidase 4 recombinant protein.

Methods: The yeast, Pichia pastoris, expression system was used for the production of the human recombinant dipeptidyl peptidase 4 as a secreted form. The full-length human dipeptidyl peptidase 4 corresponding to the amino acid 31-766 was subcloned into a Pichia pastoris expression vector, pPICZ=537;, and transformed to Pichia pastoris X33 cells.

Results: The human recombinant dipeptidyl peptidase 4 protein was expressed enzymatically as active human rDPP4_(31-766) as secreted form in the yeast P. pastoris, purified and monitored its biological activity. The test DPP4 recombinant protein induced a significant increase of DDP4 activity at 10, 20 and 30 min incubation time (p < 0.05) and at 40 min (p < 0.001). A similar pattern was found for the commercial (standard) DPP4 protein at 10, 20 and 30 min (p < 0.05) and at 40 min (p < 0.001). The high standard deviation (SD) associated with the mean value for the DPP4 activity is due to incubation time sometimes associated with high DPP4 values. The values were much higher than in other groups as expected.

Conclusion: Human recombinant dipeptidyl peptidase 4_(31-766) protein in the yeast Pichia pastoris, obtained using the technique employed in this study can further improve production efficiency and costs of human recombinant dipeptidyl peptidase 4 and other recombinant proteins.

Keywords: DPP4, Pichia pastoris, Recombinant protein, expression, Purification