Abstract
In this work, we present the production of an active 43 aa recombinant human β-defensin-1 (rhBD-1 43) in Escherichia coli AD202 cells using specific pLMM1–rhBD-1 expression system. Unique solubility properties of the C-terminal fragment of light meromyosin (LMM) allowed us to overcome foreseeable problems with isolation procedures and toxicity caused by rhBD-1 to the host organism. As a result, the majority of fusion protein (LMM–rhBD-1 43) was obtained in the soluble state, isolated by a low salt–high salt treatment of total cell protein. The rhBD-1 43 was cleaved from the fusion with Protease 4 and purified on CM Sepharose Fast Flow column with the yield of ∼1 mg rhBD-1 43 from 6 g of wet weight cells. Purified rhBD-1 43 showed antimicrobial activity against E. coli ML-35p at a concentration of 129 μM. The procedure of rhBD-1 expression and purification we present can provide a reliable and simple method for production of different cationic peptides for biological studies.
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