Construction of an Expression Vector for Production and Purification of Human Somatostatin in Escherichia coli

• Published in: Molecular biotechnology 2013-10, Vol.55 (2), p.150-158
• Main Authors: Maicas, Sergi, Moukadiri, Ismaïl, Nieto, Almudena, Valentín, Eulogio
• Format: Article
• Language: English
• Subjects: beta-galactosidase; beta-Galactosidase - genetics; beta-Galactosidase - metabolism; Biochemistry; Biological Techniques; biopharmaceuticals; Biosynthesis; Biotechnology; Cell Biology; Chemistry; Chemistry and Materials Science; Chromatography, Reverse-Phase; Cloning, Molecular; E coli; Escherichia coli; Escherichia coli - genetics; Escherichia coli - metabolism; Gene Expression; Genetic Vectors; Growth hormones; Hormones; Human Genetics; Humans; Kluyveromyces marxianus var. lactis; methionine; Methionine - metabolism; Peptides; Plasmids; Protein Science; Proteins; Recombinant Fusion Proteins - biosynthesis; Recombinant Fusion Proteins - genetics; secretion; somatostatin; Somatostatin - biosynthesis; Somatostatin - genetics; Somatostatin - isolation & purification; somatotropin
• ISSN: 1073-6085; 1559-0305
• DOI: 10.1007/s12033-013-9667-3

Somatostatin/growth hormone-inhibiting hormone is the peptide that inhibits secretion of somatotropin/growth hormone. Solid-phase synthesis methods are being currently used to produce somatostatin. Recombinant peptide synthesis is widely described for the production of small proteins and peptides; however, the production at industrial scale of peptides for biopharmaceutical applications is limited for economic reasons. Here, we propose the use of a new pGB-SMT plasmid to produce Somatostatin, as a C-terminal fusion protein with a Kluyveromyces lactis β-galactosidase fragment. To facilitate removal of that fragment by CNBr cleavage, a methionine residue was introduced at the N-terminal of the hormone peptide. The use of this construction enables an IPTG-free expression system. The suitability of this procedure has been assessed in a 15 l scale-up experiment yielding almost 300 mg, with purity >99 % and it is being implemented for commercial scale. The plasmid pGB-SMT here described is an alternative option for a cheap and high expression of other short peptide hormones.