Production of bubaline somatotropin by auto-induction in Escherichia coli.

Abstract:

:Production of His-BbST [hexahistidine-tagged BbST (bubaline somatotropin)] by auto-induction in high-density shake-flask cultures coupled with a single-step, on-column purification and refolding strategy is described here. To optimize expression of BbST, different media and expression conditions were tested. The highest expression levels of BbST, exceeding 30% of the total Escherichia coli cellular proteins, were achieved when YNG and M9NG media were used. Using these auto-inducing media, the final concentration of BbST increased up to 455 mg/l and was severalfold higher than that obtained by isopropyl beta-D-thiogalactoside induction. Most of the target protein, however, was in the form of inclusion bodies, which were solubilized in 8 M urea solution (pH 8.5). Using immobilized-metal-ion-affinity chromatography, His-BbST could be purified from solubilized sample to >97% homogeneity in a single step in a biologically active state as judged by its efficient growth-promoting activity in Nb2 rat lymphoma cell proliferation assays. The expression and purification scheme, presented here, has a potential of scaling up to obtain pure and biologically active His-BbST relatively inexpensively for further studies and applications.

authors

Sadaf S,Khan MA,Akhtar MW

doi

10.1042/BA20060154

subject

Has Abstract

pub_date

2007-05-01 00:00:00

pages

21-6

issue

Pt 1

eissn

0885-4513

issn

1470-8744

pii

BA20060154

journal_volume

47

pub_type

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