Construction of an expression vector for production and purification of human somatostatin in Escherichia coli.
Mol Biotechnol
; 55(2): 150-8, 2013 Oct.
Article
em En
| MEDLINE
| ID: mdl-23640683
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 ß-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.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Somatostatina
/
Escherichia coli
/
Vetores Genéticos
Limite:
Humans
Idioma:
En
Ano de publicação:
2013
Tipo de documento:
Article