Soluble Form Production of Recombinant Human Insulin-Like Growth Factor-1 by NusA Fusion Partner in E. coli
Trends in Peptide and Protein Sciences,
Vol. 4 (2019),
1 January 2019
,
Page 1-5 (e8)
https://doi.org/10.22037/tpps.v4i0.26948
Abstract
Insulin-Like Growth Factor-1 (IGF-1) is a small peptide with 70 amino acids and 7.6 kDa molecular weight that acts as the major mediator of growth hormone. According to the previous studies, recombinant production of human IGF-1 (rhIGF-1) in E. coli has resulted in an inactive form of protein (inclusion body). There are several strategies to transform inclusion body to a soluble form. Production in the form of fusion proteins as a suitable strategy, helps researcher in recombinant production of proteins in the soluble and active form. In current study, NusA fusion protein was used to produce IGF-1 soluble form, instead of insoluble protein. In previous study, rhIGF-1 was optimally expressed in inclusion body with 1.2 g/L concentration. rhIGF1 -NusA construct was cloned and expressed in E. coli, then, cell lysate was analyze by SDS-PAGE and densitometry techniques, to assay soluble and insoluble form of rhIGF-1. Results showed that rhIGF-1 concentration in soluble phase was 0.14 g/L, indicating that about 12% of total expression of rhIGF-1 was in the soluble form through NusA-fusion protein strategy. These results confirmed that some fusion proteins like NusA could improve the solubility of recombinant proteins expressed in heterogeneous bacterial hosts.
HIGHLIGHTS
- Fusion proteins is a suitable strategy for recombinant production of proteins in e soluble form.
- NusA fusion tag improves the solubility of recombinant proteins expressed in bacterial hosts.
- NusA fusion protein convert IGF-1 insoluble form to soluble form in E. coli.
- IGF-1
- NusA
- Soluble form
- Inclusion body
- E.coli
How to Cite
References
Babaeipour, V., Vahidi, H., Alikhani, S., Ranjbari, J., Alibakhshi, A. and M. Tabarzad, (2018). ″Effect of acyl homoserine lactone on recombinant production of human insulin-like growth factor-1 in batch culture of Escherichia coli.″ Protein and Peptide Letters, 25(11): 980-985.
Carrio, M. and A. Villaverde, (2002). ″Construction and deconstruction of bacterial inclusion bodies.″ Journal of Biotechnology, 96(1): 3-12.
Costa, S., Almeida, A., Castro, A. and L. Domingues, (2014). ″Fusion tags for protein solubility, purification and immunogenicity in Escherichia coli: the novel Fh8 system.″ Frontiers in Microbiology, 5: article 63(1-20).
Derman, A. I., Prinz, W. A., Belin, D. and J. Beckwith, (1993). ″Mutations that allow disulfide bond formation in the cytoplasm of Escherichia coli.″ Science, 262(5140): 1744-1747.
Harrison, R. (2000). ″Expression of soluble heterologous proteins via fusion with NusA protein.″ Innovations, 11: 4-7.
Lobstein, J., Emrich, C. A., Jeans, C., Faulkner, M., Riggs, P. and M. Berkmen, (2012). ″SHuffle, a novel Escherichia coli protein expression strain capable of correctly folding disulfide bonded proteins in its cytoplasm.″ Microbial Cell Factories, 11(1): article 753.
Messens, J. and J. F. Collet, (2006). ″Pathways of disulfide bond formation in Escherichia coli.″ The International Journal of Biochemistry & Cell Biology, 38(7): 1050-1062.
Ranjbari, J. (2019). ″Engineered recombinant protein production systems originated from Escherichia coli.″ Trends in Peptide and Protein Sciences, 3: 2-1-6.
Ranjbari, J., Babaeipour, V., Vahidi, H., Moghimi, H., Mofid, M. R., Namvaran, M. M. and S. Jafari, (2015). ″Enhanced production of insulin-like growth factor I protein in Escherichia coli by optimization of five key factors.″ Iranian Journal of Pharmaceutical Research: IJPR, 14(3): 907-917.
Raran-Kurussi, S. and D. S. Waugh, (2012). ″The ability to enhance the solubility of its fusion partners is an intrinsic property of maltose-binding protein but their folding is either spontaneous or chaperone-mediated.″ PloS One, 7(11): e49589.
Rosano, G. L. and E. A. Ceccarelli, (2014). ″Recombinant protein expression in Escherichia coli: advances and challenges.″ Frontiers in Microbiology, 5: article 172 (1-17).
Taylor, S. C., Berkelman, T., Yadav, G. and M. Hammond, (2013). ″A defined methodology for reliable quantification of Western blot data.″ Molecular Biotechnology, 55(3): 217-226.
Terpe, K. (2003). ″Overview of tag protein fusions: From molecular and biochemical fundamentals to commercial systems.″ Applied Microbiology and Biotechnology, 60(5): 523-533.
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