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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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Abstract

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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Authors and Affiliations

  1. Marine Biotechnology and Ecology Discipline, Central Salt and Marine Chemicals Research Institute, 364 002, Bhavnagar, Gujarat, India

    Yasmin Khambhaty, Kalpana Mody & B. Jha

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  1. Yasmin Khambhaty
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  2. Kalpana Mody
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  3. B. Jha
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Correspondence to Kalpana Mody.

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Khambhaty, Y., Mody, K. & Jha, B. Purification and characterization of κ-carrageenase from a novel γ-proteobacterium,Pseudomonas elongata (MTCC 5261) syn.Microbulbifer elongatus comb. Nov.. Biotechnol. Bioprocess Eng. 12, 668–675 (2007). https://doi.org/10.1007/BF02931084

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