Use of Ssp dnaB mini-intein as a fusion partner for preparation of recombinant human brain natriuretic peptide / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 180-203 passim, 2006.
Article
in Chinese
| WPRIM
| ID: wpr-237004
ABSTRACT
Human brain natriuretic peptide (hBNP) was used clinically for the treatment of acute decompensated congestive heart failure. In this paper, hBNP was expressed as a fusion protein with a histidine tag and Ssp dnaB mini-intein which was capable of self-cleavage. After affinity chromatography with Ni-Sepharose and renaturation, the fusion protein was enriched with CM-cellulose. Ssp dnaB mini-intein mediated peptide-bond hydrolysis was triggered by shifting the pH and temperature in the CM-cellulose column, which let to the release of hBNP from the fusion protein and the separation of hBNP from His-DnaB. The hBNP sample was further purified by C4 reverse phase HPLC, and 2.8mg of the peptide with homogeneity of 97% was obtained from one liter of culture medium. The biological activity was assayed in vitro, which indicated that hBNP had a potent vasodilatory effect on rabbit aortic strips with an EC50 of 1.94 x 10(-6) mg/mL.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Pharmacology
/
Recombinant Fusion Proteins
/
Protein Engineering
/
Protein Splicing
/
Natriuretic Peptide, Brain
/
Inteins
/
Escherichia coli
/
DnaB Helicases
/
Genetic Vectors
/
Genetics
Limits:
Animals
Language:
Chinese
Journal:
Chinese Journal of Biotechnology
Year:
2006
Type:
Article
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