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Isolation, evaluation and purification of antibacterial peptides from rhinoceros beetle, Oryctes rhinoceros

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Author(s): M. Rabeeth, Muthukumar M., Balaji Rajkumar M and and Kesava Kumar H.

Journal: GERF Bulletin of Biosciences
ISSN 2229-6433

Volume: 3;
Issue: 2;
Start page: 16;
Date: 2012;
Original page

Keywords: Antibacterial peptide | Oryctes rhinoceros | Tricine SDS-PAGE | Antibacterial activity | MIC | Gel filtration | RP-HPLC

ABSTRACT
Antibacterial peptides were isolated from immune haemolymph of Rhinoceros beetle (Oryctes rhinoceros L.) larvae and evaluated for antibacterial activity against bacterial pathogens, viz., Klebsiella pneumoniae, Micrococcus luteus, Bacillus subtilis, Paenebacillus macerans and Pseudomonas sp. The peptides were found to be stable upto 37°C and antibacterial activity observed at pH 7.0. The peptide were active against all the pathogens with minimum inhibitory concentrations varying between 1.56 to 6.25 μM. Protein profiling on 16% Tricine sodium dodecyl sulphate polyacrylamide gel revealed that the molecular size of two antimicrobial peptides of 17 and 9 kDa, which was also observed as two peak fractions from reverse phase high pressure liquid chromatography (RP-HPLC) exhibiting antibacterial activity. These peptides were purified by Gel filtration chromatography and RP-HPLC upto 15 and 37 fold purifications, respectively.
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