Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
8360200 | Protein Expression and Purification | 2015 | 7 Pages |
Abstract
Recombinant human B-type natriuretic peptide (rhBNP) is a 32-amino acid peptide used to treat congestive heart failure. In this paper, we report a method for the increased production of rhBNP in Escherichia coli with high purity. hBNP was cloned with a short growth hormone fusion partner coupled with a unique acid-labile dipeptide linker to cleave the fusion protein to release the rhBNP. The recombinant fusion protein was expressed as an inclusion body (IB) and the fermentation process was optimized to produce on large scale. The IBs were recovered by cell lysis, and the pure IBs were directly treated with diluted acid to get the target peptide from the fusion protein and the resultant peptide was purified by reversed phase chromatography. The final purity of the rhBNP was more than 99% with yield of 50Â mg per liter of culture, which is ten times higher than the previous reports. The purified rhBNP exhibited specific biological activity similar to the standard peptide in producing cyclic-guanosine monophosphate.
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Biochemistry
Authors
Radha Madhavi Kanumuri, Chitra Bajji, Rajesh R. Tummuru, Venkat R.R. Arva Tatireddigari, Lakshmi Narasu Mangamoori, Kalpana Panati, Venkata Ramireddy Narala,