Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
10843372 | Protein Expression and Purification | 2010 | 7 Pages |
Abstract
The recombinant proteins were expressed in Escherichia coli BL21 (DE3) and solubilized with Triton X-100. The purification was achieved using cellulose CF11 fibers, taking advantage of the CBM3 specific affinity for cellulose; after hydrolysis with formic acid, LL37 was further purified by reverse-phase HPLC, as confirmed by MALDI-TOF mass spectrometry. The production and purification methodology developed in this work compares advantageously to other protocols previously described, having fewer purification steps. Only the recombinant LL37 obtained from the C-terminally fused protein (LK-CBM3-LL37) showed antibacterial activity against E. coli K12, with a MIC of 180 μg/ml.
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Authors
Reinaldo Ramos, LucÃlia Domingues, Miguel Gama,