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KMID : 0613820080180060789
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Journal of Life Science
2008 Volume.18 No. 6 p.789 ~ p.795
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Purification of Vibrio anguillarum Growth Inhibition Factor Produced by Bacillus amyloliquefaciens H41
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Shin Hyun-Chul
Chung Kyung-Tae
Kim Kwang-Hyeon
Kim Byung-Woo
Kwon Hyun-Ju
Lee Eun-Woo
Yum Jong-Hwa
Rhu Eun-Ju
Jeong Yu-Jeong
Kim Young-Hee
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Abstract
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To study the possible use of probiotics in fish farming, we evaluated antagonism of antibacterial strain Bacillus amyloliquefaciens H41 against the fish pathogenic bacterium Vibrio anguillarum NCMB1. The purification of growth inhibition factor produced by B. amyloliquefaciens H41 was achieved by obtaining supernatant of this bacterium. The growth inhibition factor was purified to homogeneity by 70% ammonium sulfate precipitation, DEAE-sephadex A-50 ion exchange chromatography, sephadex G-200 gel filtration column chromatography, and sephadex G-50 gel filtration column chromatography with 40.8 fold of purification and 2.9% yield. The molecular weight of the purified growth inhibition factor was 48 kDa as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The optimum pH and temperature for the growth inhibition factor were pH 7.5 and 30℃, respectively. The activity of growth inhibition factor was enhanced slightly by some metal ions, such as Mg?², Mn?², but was inhibited by the addition of Co?², Hg?², Zn?² and Ag?². NaCl stability of the growth inhibition factor was observed with 50% residual activity at 3% NaCl concentration. Toxicity test showed that the purified B. amyloliquefaciens H41 growth inhibition factor did not affect the live of Japanese flounder (Paralichthys olivaceus) and the effectiveness was 78% of residual lethality compared to commercial antibacterial agents.
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KEYWORD
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B.amyloliquefaciensH41, growthinhibitionfactor, Vibrioanguillarum
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