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Enhanced production and purification of recombinant surface array protein (Sap) for use in detection of Bacillus anthracis

Author:
Puranik, Nidhi, Tripathi, N.K., Pal, V., Goel, AjayKumar
Source:
3 Biotech 2018 v.8 no.5 pp. 254
ISSN:
2190-572X
Subject:
Bacillus anthracis, Escherichia coli, Western blotting, affinity chromatography, antibodies, bacteria, batch fermentation, biomarkers, culture flasks, culture media, polyacrylamide gel electrophoresis
Abstract:
Surface array protein (Sap) can be an important biomarker for specific detection of Bacillus anthracis, which is released by the bacterium during its growth in culture broth. In the present work, we have cloned and expressed Sap in Escherichia coli. The culture conditions and cultivation media were optimized and used in batch fermentation process for scale up of Sap in soluble form. The recombinant Sap was purified employing affinity chromatography followed by diafiltration. The final yield of purified protein was 20 and 46 mg/l of culture during shake flasks and batch fermentation, respectively. The protein purity and its reactivity were confirmed employing SDS–PAGE and Western blot, respectively. The antibodies raised against purified Sap were evaluated by Western blotting for detection of Sap released by B. anthracis. Our results showed that the Sap could be a novel marker for detection and confirmation of B. anthracis.
Agid:
5967574