The tandem repeats of LFB15(W4,10)-HP(4-16) (LH) gene were cloned into vector pET32a(+) for recombinant expression in Escherichia coli. The E. coli C43(DE3) was successfully used as the expression host to avoid the cell death during induction in E. coli BL21(DE3). Fusion LH dimer was expressed as inclusion body at a portion of 35% of total cell protein and could be well purified by Ni2+-chelating chromatography. The recombinant LH was released by the cleavage of 50% formic acid, and its yield reached 11.3 mg/l with purity of 95%. The MIC50 of 3.6 and 1.9 μM of recombinant LH against E. coli CMCC 44102 and Bacillus subtilis ATCC 6633 were determined, respectively. The results demonstrated that expression of tandem LH gene in E. coli C43(DE3) and formic acid cleavage would provide a potent efficient platform for the production of interested peptides.
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This study is supported by National Natural Science Foundation of China (no. 30771574; no. 30810303084), Beijing Natural Science Foundation (no. 5062031; no. 5093030) and Chinese 863 Program (no. 2004AA246040).
Zi-gang Tian and Tian-tang Dong contributed equally to this paper.
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Tian, Z., Dong, T., Yang, Y. et al. Expression of antimicrobial peptide LH multimers in Escherichia coli C43(DE3). Appl Microbiol Biotechnol 83, 143–149 (2009). https://doi.org/10.1007/s00253-009-1893-z
- Antimicrobial peptide
- Escherichia coli
- Formic acid