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KMID : 1094720070120060668
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Biotechnology and Bioprocess Engineering
2007 Volume.12 No. 6 p.668 ~ p.675
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Purification and characterization of κ-carrageenase from a novel γ-proteobacterium, Pseudomonas elongata (MTCC 5261) syn. Microbulbifer elongatus comb. Nov.
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Khambhaty Yasmin
Mody Kalpana
Jha Bhavanath
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Abstract
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The phenotypic and carrageenolytic features of a novel halo tolerant marine bacterium, isolated from decayed red algal samples collected along the west coast of India were studied. This gram-negative strain was identified asPseudomonas elongala (MTCC 5261) syn.Microbulbifer elongalus comb. nov according to its morphological, physiological and molecular characterization. The extracellular κ-carrageenase was purified 106.54-fold by a combination of ammonium sulfate precipitation (40∼60%) and successive gel filtration chromatography. The purified protein fraction yielded significantly high activity of 426.19 units/mg protein and migrated as a single band on a sodium dodecyl sulfate polyacrylamide gel electrophoresis with a molecular weight of ∼128 kDa. For κ-carrageenase activity, optimum temperature was 40°C whereas two pH optimai.e. 5.6 and 7.7 were observed. For κ-carrageenan, the enzyme gave aK m value of 6.66 mg/mL and aV max value of 4 μmol/min/mg when the reaction was carried out at 40°C and pH 5.6. Isolated κ-carrageenase could successfully generate protoplasts ofKappaphycus alvarezii. This is the first report on the production of κ-carrageenase by this bacterium isolated from west coast of India. Molecular mass and various characteristics showed that the carrageenase fromP. elongata was much different from those previously reported.
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KEYWORD
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κ-carrageenan, κ-carrageenase, Kappaphycus alvarezii, protoplast isolation, Pseudomonas elongata
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