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KMID : 1094720080130050584
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Biotechnology and Bioprocess Engineering
2008 Volume.13 No. 5 p.584 ~ p.591
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Purification, characterization and application of a novel extracellular agarase from a marine Bacillus megaterium
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Khambhaty Yasmin
Mody Kalpana
Jha Bhavanath
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Abstract
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A marine, gram positive, aerobic, spore forming, and non flagellated bacterium which degrades low melting point (LMP) ? agarose was isolated from the west coast of India and identified as Bacillus megaterium based on its morphological, biochemical, and molecular characterization. This bacterium produced clear haloes or zone of clearance on agar containing plates which was a clear indication of its agarolytic property. The extracellular agarase thus obtained was purified 8.8 and 78 fold from the culture supernatant by ammonium sulfate precipitation and gel filtration, respectively. Molecular mass by gel filtration and SDS-PAGE gave values of 15 and 12 kDa, respectively. The optimum temperature and pH for maximum agarase activity were 40°C and 6.6. The activity of agarase was drastically reduced by addition of metal ions in the assay system. This agarase, gave a K m and V max value of 4 mg/mL and 2.75 μmol/min/mg. The isolation of protoplast from agarophyte like Gelidiella acerosa using indegenous agarase is reported for the first time.
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KEYWORD
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agarase, Bacillus megaterium, Gelidiella acerosa, low melting point agarose, red algal protoplast
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