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KMID : 0613820090190070994
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Journal of Life Science
2009 Volume.19 No. 7 p.994 ~ p.1002
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Characterization and Purification of the Bacteriocin Produced by Bacillus licheniformis Isolated from Soybean Sauce
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Jung Sung-Sub
Choi Jung-I
Joo Woo-Hong
Suh Hyun-Hyo
Na Ae-Sil
Cho Yong-Kweon
Moon Ja-Young
Ha Kwon-Chul
Paik Do-Hyun
Kang Dae-Ook
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Abstract
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A bacteriocin-producing bacterium identified as Bacillus licheniformis was isolated from soybean sauce. Antibacterial activity was confirmed by paper disc diffusion method, using Micrococcus luteus as a test organism. The bacteriocin also showed antibacterial activities against Bacillus sphaericus, Lactobacillus bulgaricus, Lactobacillus plantarum, Paenibacillus polymyxa, and Pediococcus dextrinicus. Optimal culture conditions for the production of bacteriocin was attained by growing the cells in an MRS medium at a pH of 6.5～7.0 and a temperature of 37℃ for 36～48 hr. Solvents such as chloroform, ethanol, acetone, and acetonitrile had little effect on bacteriocin activity. However, about 50% of bacteriocin activity diminished with treatment of methanol and isopropanol at the final concentration of 50% at 25℃ for 1 hr. It was stable against a pH variation range from 3.0 and 7.0, but the activity reduced to 50% at a pH range from 9.0 to 11.0. It’s activity was not affected by heat treatment at 100oC for 30 min and 50% of activity was retained after heat treatment at 100℃ for 60 min, showing high thermostability. The bacteriocin was purified to a homogeneity through ammonium sulfate precipitation, SP-Sepharose ion-exchange chromatography, and reverse-phase high-performance liquid chromatography (HPLC). The entire purification protocol led to a 75-fold increase in specific activity and a 13.5% yield of bacteriocin activity. The molecular weight of purified bacteriocin was estimated to be about 2.5 kDa by tricine-SDS-PAGE.
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KEYWORD
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Bacteriocin, dic diffusion method, antibacterial spectrum, purification, stability, soybean sauce
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