Biological Activity of Persian Sturgeon Recombinant Growth Hormone Molecules Trapped In Inclusion Bodies (IBs)

Document Type : Research Articles

Authors

• Ehsan Nasr ¹
• Hrachya Hovhannisyan ²
• Pourkazemi Mohammad ³

¹ Islamic Azad University of meshkin Shahr

² Yerevan Vniversity

³ Rasht

https://doi.org/10.22067/jcmr.v9i1.53803

Abstract

The biological activity of Persian sturgeon (PS) enfolded crude recombinant growth hormone (rGH) recovered from inclusion bodies (IB) estimated by administrating to fish fingerlings. The psrGH IBs expressed in E. coli were dissolved in cold guanidine HCl solution, immediately diluted ~1000 fold in cold sodium chloride 0.9% solution do not allow the rGH folding and administrated to fishes. It was revealed that intramuscularly injections of rGH at dosage of 0.5 μg/ g once a week for 8 weeks significantly accelerate the growth of fishes. Comparison of the mean growth rate and daily weight and length gain dates of Persian sturgeon fingerlings administrated by pure and crude grade rGH revealed the existence at list 2-5% potentially biologically active psrGH molecules in IBs. This fact enables to avoid the time and cost consuming processes of refolding and purification of rGHs.

Keywords

• Keywords: Inclusion body
• Persian sturgeon
• Recombinant growth hormone
• Biological activity

20.1001.1.20089147.2017.9.1.7.2

References

1.Baneyx F. and Mujacic M. (2004) Recombinant protein folding and misfolding in Escherichia coli. Nature Biotechnology. 22: 1399 – 1408.

2.Barannikova, I. A. (1987) Review of sturgeon farming in the Soviet Union. Journal of Ichthyology, 27: 62–67.

3.De Bernardez C.E. (1998) Refolding of recombinant proteins. Current Opinion in Biotechnology, 9: 157–163.

4.Doglia S., Ami D., Natalello A., Gatti-Lafranconi P.and Lotti M. (2008) Fourier transform infrared spectroscopy analysis of the conformational quality of recombinant proteins within inclusion bodies. Biotechnology Journal,3: 193-201.

5.Garcia-Fruitos E., Aris A. and Villaverde A. (2007) Localization of functional polypeptides in bacterial inclusion bodies. Applied and Environmental Microbiology, 73: 289-294.

6.Garcia-Fruitos E. Martinez-Alonso M., Gonzalez-Montalban N., Valli M., Mattanovich D., and Villaverde A. (2007) Divergent genetic control of protein solubility and conformational quality in Escherichia coli. Journal of Molecular Biology, 374: 195-205.

7.Jevsevar S., Gaberc-Porekar V., Fonda I., Podobnik B., Grdadolnik J. and Menart V. (2005) Production of nonclassical inclusion bodies from which correctly folded protein can be extracted. Biotechnology Progress, 21: 632–639.

8.Lilie H., Schwarz E., and Rudolph R. (1998) Advances in refolding of proteins produced in E. coli. Curr. Opin. Biotechnology, 9: 497–501.

9.Marston F.A.O. (1986) The purification of eukaryotic polypeptides synthesized in Escherichia coli. Biochemical Journal, 240(1): 1-12.

10.Misawa S. and Kumagai I. (1999) Refolding of therapeutic proteins produced in Escherichia coli as inclusion bodies. Biopolymers, 51: 297-307.

11.Rudolph R. and Lilie H. (1996) In vitro folding of inclusion body proteins. The Journal of Federation of American Societies for Experimental Biology, 10: 49-56.

12.Schein C.H. (1989) Production of Soluble Recombinant Proteins in Bacteria. Nature Biotechnology, 7: 1141 – 1149.

13.Singh S.M. and Panda A.K (2005) Solubilization and refolding of bacterial inclusion body proteins. Journal of Bioscience and Bioengineering, 99: 303–310.

14.Ventura S. and Villaverde A. (2006) Protein quality in bacterial inclusion bodies.Trends Biotechnol. 24(4): 179-185.