title: Construction of an expression vector for production and purification of human somatostatin in Escherichia coli.
abstract: 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.
authors: Departament de Microbiologia i Ecologia, Universitat de València, Dr. Moliner, 50, 46100, Burjassot, Spain, sergi.maicas@uv.es.
authors: Sergi
authors: S
authors: Maicas
authors: Ismaïl
authors: I
authors: Moukadiri
authors: Almudena
authors: A
authors: Nieto
authors: Eulogio
authors: E
authors: Valentín
mesh_terms: D056148:Chromatography, Reverse-Phase; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D008715:Methionine / Q000378:metabolism; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D013004:Somatostatin / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification*; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
chemical_list: D011993:Recombinant Fusion Proteins; D013004:Somatostatin; D008715:Methionine; D001616:beta-Galactosidase
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