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Original Research Article
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
1Department of Medical
Laboratory Science, College of Health Science, Dankook
University, Cheonan-si, Chungnam, 330-714, Republic of
Korea
*For correspondence:
Email:
hyunbio@gmail.com,
hkang@dankook.ac.kr; Tel:
82-41-550-1452; Fax: 82-41-559-7934
Received: 6 April 2014
Revised accepted: 10 May
2014
Tropical Journal of Pharmaceutical Research, June
2014; 13(6): 855-862
http://dx.doi.org/10.4314/tjpr.v13i6.5
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, 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 |
