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Production of Extremely Alkaliphilic, Halotolerent, Detergent, and Thermostable Mannanase by the Free and Immobilized Cells of Bacillus halodurans PPKS-2. Purification and Characterization

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Abstract

The alkaliphilic Bacillus halodurans strain PPKS-2 was shown to produce extracellular extreme alkaliphilic, halotolerent, detergent, and thermostable mannanase activity. The cultural conditions for the maximum enzyme production were optimized with respect to pH, temperature, NaCl, and inexpensive agro wastes as substrates. Mannanase production was enhanced more than 4-fold in the presence of 1 % defatted copra meal and 0.5 % peptone or feather hydrolysate at pH 11 and 40 °C. Mannanase was purified to 10.3-fold with 34.6 % yield by ion exchange and gel filtration chromatography methods. Its molecular mass was estimated to be 22 kDa by SDS-PAGE. The mannanase had maximal activity at pH 11 and 70 °C. This enzyme was active over a broad range of NaCl (0–16 %) and thermostable retaining 100 % of the original activity at 70 °C for 3 h. Immobilization of whole cells proved to be effective for continuous production of mannanase. Since the strain PPKS-2 grows on cheaper agro wastes such as defatted copra meal, corn husk, jowar bagasse, and wheat bran, these can be exploited for mannanase production on an industrial scale.

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Acknowledgments

We thank UGC, New Delhi, for providing grants for JRF under UGC-SAP (DSR-I) scheme to VS.

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Authors and Affiliations

  1. Department of Biochemistry, Gulbarga University, Gulbarga, 585106, Karnataka, India

    S. Vijayalaxmi, V. H. Mulimani & K. Sreeramulu

  2. Department of Biochemistry, The Oxford College of Science, Bangalore, 560102, Karnataka, India

    P. Prakash

  3. Agricultural Research Station, University of Agricultural Sciences, Gulbarga, 585103, Karnataka, India

    S. K. Jayalakshmi

Authors
  1. S. Vijayalaxmi
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  2. P. Prakash
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  3. S. K. Jayalakshmi
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  4. V. H. Mulimani
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  5. K. Sreeramulu
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Correspondence to K. Sreeramulu.

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Vijayalaxmi, S., Prakash, P., Jayalakshmi, S.K. et al. Production of Extremely Alkaliphilic, Halotolerent, Detergent, and Thermostable Mannanase by the Free and Immobilized Cells of Bacillus halodurans PPKS-2. Purification and Characterization. Appl Biochem Biotechnol 171, 382–395 (2013). https://doi.org/10.1007/s12010-013-0333-9

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