Purification of recombinant human growth hormone produced in soluble form in Escherichia coli in lab-scale

Subject Areas : Industrial Microbiology

Seyed Morteza  Robatjazi ¹ (Assistant Professor, Malek Ashtar University of Technology, Faculty of Chemistry and Chemical Engineering, Department of Bioscience and Biotechnology.)
Seyed Mohammad  Hasanpour Matikolaee ² (M.Sc., student, Malek Ashtar University of Technology, Faculty of Chemistry and Chemical Engineering, Department of Bioscience and Biotechnology,)
Valiolah Babaeipour  Babaeipour ³ (Associate Professor, Malek Ashtar University of Technology, Faculty of Chemistry and Chemical Engineering, Department of Bioscience and Biotechnology,)
Hamideh  Rouhani Nejad ⁴ (Assistant Professor, Malek Ashtar University of Technology, Faculty of Chemistry and Chemical Engineering, Department of Bioscience and Biotechnology.)

Received: 2019-08-27 Accepted : 2019-10-22 Published : 2020-02-20

Keywords: Recombinant human growth hormone (rhGH), Cytoplasmic expression, Affinity chromatography,

Abstract :

Background & Objectives: Human growth hormone (hGH) or somatotropin is a single chain polypeptide that consisting of 191 amino acid residues and a molecular weight of 22kDa. The purpose of this study was to evaluate the production and purification of the soluble form of recombinant human growth hormone in the lab-scale.Material & Methods: In this study, the recombinant E. coli containing the plasmid pET32a(+)-hGH was used. To obtain high-level production of soluble hGH in the cytoplasm was used from Trx tag and for the purification process of hGH was used from His tag. The bacteria were grown in LB and TB culture media at 25 ⁰C. The purification process carried out based on affinity chromatography using Ni-NTA resin.Results: The SDS-PAGE analysis showed 55% hGH expression that 33.7% expressed as soluble and 18.8% expressed as IBs. The amount of hGH fusion protein purified through a Ni-NTA column was 22. 4 mg per gram of wet cells.Conclusion: The results of this study indicate that hGH-Trx fusion protein was properly expressed. The purity of hGH fusion protein purified by the histidine-tagged protein purification method was determined by 91% with a total yield of 38%.

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