Zahra Hajihassan Seyed Kazem Hosseini Alireza Zomorodipour


Human activin A is a member of the transforming growth factor-β superfamily consists of two similar beta subunits. Activin A is expressed by different cells and displays numerous biological activities such as control of neuronal cell proliferation and differentiation, promotion of neuronal survival in the body. Therefore, recombinant production of activin A is beneficial because it can be used to treat many neurodegenerative diseases such as Alzheimer's and Parkinson diseases. In this study E. coli as a cheap and fast-growing host was selected to produce recombinant human activin A. As cytoplasmic expression of human activin A with complex structure and disulfide bonds produces inclusion bodies, so periplasmic expression of it can be beneficial. Therefore, we used modified Iranian B. licheniformis α-amylase signal peptide as a new signal peptide in order to translocate the recombinant activin A through the inner membrane. In this study human pro-activin A cDNA and signal sequence were cloned in pET21b vector and resulting vector transformed into the two strains of E. coli BL21. SDS-PAGE and western blot techniques were used to confirm recombinant activin A expression. Finally, our results indicated that the signal peptide used in this study was effective for secretion of activin A into the periplasmic space of E. coli.

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Berkmen M. (2012) Production of disulfide-bonded proteins in Escherichia coli. Protein Expr Purif. 82(1): 240-251.

Chen YG., Wang Q., Lin SL., Chang CD., Chuang J. and Ying SY. (2006) Activin signaling and its role in regulation of cell proliferation, apoptosis, and carcinogenesis. Exp Biol Med. 231(5): 534-44.

Choi JH. and Lee SY. (2004) Secretory and extracellular production of recombinant proteins using Escherichia coli. Appl Microbiol Biotechnol. 64(5): 625-635.

Cronin CN., Thompson DA. and Martin F. (1998) Expression of bovine activin-A and inhibin-A in recombinant baculovirus-infected Spodoptera frugiperda Sf 21 insect cells. Int J Biochem Cell Biol. 30(10): 1129-45.

Demaio A. (1996) Protein blotting and immunoblotting using nitrocellulose membranes. In protein blotting. Oxford University Press. Oxford. Pp. 11-30.

Hajihassan Z., Sohrabi M., Rajabi Bazl M. and Eftekhary HS. (2016) Expression of Human Nerve Growth Factor beta and Bacterial Protein Disulfide Isomerase (DsbA) as a Fusion Protein (DsbA:: hNGF) Significantly Enhances Periplasmic Production of hNGF beta in Escherichia coli. Romanian Biotechnological Letters. 21(5): 11850-11856.

Han S., Machhi S., Berge M., Xi G., Linke T. and Schoner R. (2017) Novel signal peptides improve the secretion of recombinant Staphylococcus aureus Alpha toxinH35L in Escherichia coli. AMB Express. 7(1):93.

Laemmli UK. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 227 (5259): 680-685.

Libby RT., Braedt G., Kronheim SR., et al. (1987) Expression and purification of native human granulocyte-macrophage colony-stimulating factor from an Escherichia coli secretion vector. DNA. 6(3): 221-229.

Low KO., Muhammad Mahadi N. and Md Illias R. (2013) Optimisation of signal peptide for recombinant protein secretion in bacterial hosts. Appl Microbiol Biotechnol. 97(9): 3811-26.

Low KO., Muhammad Mahadi N. and Md Illias R. (2013) Optimisation of signal peptide for recombinant protein secretion in bacterial hosts. Appl Microbiol Biotechnol. 97(9): 3811-26.

Natale P., Brüser T. and Driessen AJ. (2008) Sec-and Tat-mediated protein secretion across the bacterial cytoplasmic membrane-distinct translocases and mechanisms. Biochim Biophys Acta. 1778(9): 1735-1756.

Pangas SA. and Woodruff TK. (2002) Production and purification of recombinant human inhibin and activin. J Endocrinol. 172: 199–210.

Papakonstantinou T., Harris SJ., Fredericks D., Harrison C., Wallace EM. and Hearn MT. (2009) Synthesis, purification and bioactivity of recombinant human activin A expressed in the yeast Pichia pastoris. Protein Expr Purif. 64(2): 131-138.

Rosano GL. and Ceccarelli EA. (2014) Recombinant protein expression in Escherichia coli: advances and challenges. Front Microbiol. 5: 172.

Sahdev S., Khattar SK. and Saini KS. (2008) Production of active eukaryotic proteins through bacterial expression systems: a review of the existing biotechnology strategies. Mol Cell Biochem. 307: 249-264.

Sambrook J. and Russel DW. (2001) Molecular cloning: A laboratory manual. 4th edition. New York, USA. Cold spring harbor laboratory press.

Schubert D., Kimura H., LaCorbiere M., Vaughan J., Karr D. and Fischer WH. (1990) Activin is a nerve cell survival molecule. Nature. 344: 868-870.

Sulyok S., Wankell M., Alzheimer C. and Werner S. (2004) Activin: an important regulator of wound repair, fibrosis, and neuroprotection. Mol Cell Endocrinol. 225(1–2): 127–32.

Tegel H., Ottoson J. and Hober S. (2011) Enhancing the protein production levels in Escherichia coli with a strong promoter. The FEBS J. 278 (5): 729-739.

Walton KL., Makanji Y. and Harrison CA. (2012) New insights into the mechanisms of activin action and inhibition. Mol Cell Endocrinol. 359(1-2): 2-12.

Weiss A. and Attisano L. (2013) The TGFbeta superfamily signaling pathway. Wiley Interdiscip Rev Dev Biol. 2(1):47-63.

Zamani M., Nezafat N., Negahdaripour M., Dabbagh F. and Ghasemi Y. (2015) In Silico Evaluation of Different Signal Peptides for the Secretory Production of Human Growth Hormone in E. coli. INT J PEPT RES THER. 21(3): 261-268.
How to Cite
Zahra, H., Seyed kazem, H., & Alireza, Z. (2018). A Novel Signal Peptide Derived from Bacillus Licheniformis α-Amylase Efficiently Targets Recombinant Human Activin A to the Periplasm of Escherichia coli. Journal of Cell and Molecular Research, 10(1), 47-51. https://doi.org/10.22067/jcmr.v8i2.64629
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