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Production of alkaliphilic, halotolerent, thermostable cellulase free xylanase by Bacillus halodurans PPKS-2 using agro waste: single step purification and characterization

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Abstract

The alkaliphilic Bacillus halodurans strain PPKS-2 was shown to produce extracellular alkaliphilic, thermostable and halotolerent xylanase. The culture conditions for xylanase production were optimized with respect to pH, temperature, NaCl and inexpensive agro waste as substrates. Xylanase yield was enhanced more than four fold in the presence of 1% corn husk and 0.5% peptone or feather hydrolysate at pH 11 and 37°C. Xylanase was purified to 11.8-fold with 8.7% yield by using traditional chromatographic methods whereas the same enzyme purified to 20-fold with 72% yield by using corn husk as ligand. Its molecular mass was estimated to be 24 kDa by SDS–PAGE. The xylanase had maximal activity at pH 11 and 70°C. The enzyme was active over broad range, 0–20% sodium chloride. The enzyme was thermostable retaining 100% of the original activity at 70°C for 3 h. The apparent K m values for oat spelt xylan and brichwood xylan were 4.1 and 4.4 mg/ml respectively. The deduced internal amino acid sequence of PPKS-2 xylanase resembled the sequence of β-1,4-endoxylanase, which is member of glycoside hydrolase family 11.

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Acknowledgments

This research was supported by a grant to K.S. from the University Grants Commission and UGC-SAP (DRS-I), New Delhi.

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Authors and Affiliations

  1. Agricultural Research Station, University of Agricultural Sciences, Raichur, Gulbarga, 585103, India

    P. Prakash & S. K. Jayalakshmi

  2. Department of Biological Sciences, Tata Institution of Fundamental Research (TIFR), Mumbai, 400005, India

    B. Prakash & M. Rubul

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

    P. Prakash & K. Sreeramulu

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  1. P. Prakash
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  2. S. K. Jayalakshmi
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  3. B. Prakash
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  5. K. Sreeramulu
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Correspondence to K. Sreeramulu.

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Prakash, P., Jayalakshmi, S.K., Prakash, B. et al. Production of alkaliphilic, halotolerent, thermostable cellulase free xylanase by Bacillus halodurans PPKS-2 using agro waste: single step purification and characterization. World J Microbiol Biotechnol 28, 183–192 (2012). https://doi.org/10.1007/s11274-011-0807-2

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