Expression of a modified antimicrobial peptide BhSGAMP-1-S (Bradysia hygida) in Escherichia coli and characterization of its activity
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Supported by the National Natural Science Foundation of China (30872033) and the Program for New Century Excellent Talents in University (NECT-06-0867)

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    Abstract:

    bstract: [Objective] Bradysia hygida salivary glands antimicrobial peptide 1 (BhSGAMP-1) is one of the antimicrobial peptides involved in a preventive mechanism of defense of the fly Bradysia hygida. To know better about the molecular characterization of this antimicrobial peptide, we expressed and purified the modified BhSGAMP-1-S and investigated its antimicrobial activity. [Methods] We synthesized the gene of BhSGAMP-1-S designed with preferred codons of Escherichia coli and expressed it as a fusion protein in E.coli TB1 by using pMAL-c2X as vector. We purified the fusion protein using amylase resin affinity chromatography. In addition, we cleaved the fusion protein by enterokinase and the released recombinant BhSGAMP-1-S was separated by size exclusion chromatography, then followed by reversed-phase high performance liquid chromatography. We analyzed the antimicrobial activities of the purified recombinant BhSGAMP-1-S by bioassay. [Results] The fusion protein was mostly expressed in soluble form under the optimized conditions. The recombinant BhSGAMP-1-S was produced with a pure yield of 0.38 mg/100 mL culture medium. Antimicrobial assays demonstrated that the recombinant BhSGAMP-1-S was active against several Gram-positive and Gram-negative bacteria and fungi. [Conclusion] It appears to be first successful production of the recombinant BhSGAMP-1-S from fly Bradysia hygida. Data presented here confirm that the recombinant BhSGAMP-1-S is now ready for further studying and characterize their antimicrobial properties.

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Jue Wang, Qiurong Gao, Qiang Liu, Linping Xiang, Yuqin Wang, Dun Wang. Expression of a modified antimicrobial peptide BhSGAMP-1-S (Bradysia hygida) in Escherichia coli and characterization of its activity. [J]. Acta Microbiologica Sinica, 2010, 50(9): 1185-1193

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History
  • Received:March 24,2010
  • Revised:April 20,2010
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