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Purification and characterization of cellulase-free low molecular weight endo β-1,4 xylanase from an alkalophilic Cellulosimicrobium cellulans CKMX1 isolated from mushroom compost

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Abstract

Alkalophilic Cellulosimicrobium cellulans CKMX1 isolated from mushroom compost is first report on actinomycetes that has the ability to produce thermostable cellulase-free xylanase, which is an important industrial enzyme used in the pulp and paper industry. Strain CKMX1 was characterized by metabolic fingerprinting, whole-cell fatty acids methyl ester analysis and 16Sr DNA and found to be C. cellulans CKMX1.The enzyme was purified by gel permeation and anion exchange chromatography and had a molecular mass of 29 kDa. Xylanase activity was optimum at pH 8.0 and 55 °C. The enzyme was somewhat thermostable, retaining 50 % of the original activity after incubation at 50 °C for 30 min. The xylanase had K m and V max values of 2.64 mg/ml and 2,000 µmol/min/mg protein in oat spelt xylan, respectively. All metal ions except HgCl2, CoCl2 as well as CdCl2 were well tolerated and did not adversely affect xylanase activity. The deduced internal amino acid sequence of C. cellulans CKMX1 xylanase by matrix assisted laser desorption ionization-time of flight mass spectrometry resembled the sequence of β-1,4-endoxylanase, which is a member of glycoside hydrolase family 11. Some of the novel characteristics that make this enzyme potentially effective in xylan biodegradation could be useful for pulp and paper biobleaching are discussed in this manuscript.

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Acknowledgments

We thank Department of Science and Technology, Ministry of Science and Technology, Govt. of India, for providing contingency grant through Inspire Fellowship.

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Authors and Affiliations

  1. Department of Basic Sciences (Microbiology Section), Dr. Y. S. Parmar University of Horticulture and Forestry, Nauni, Solan, 173230, H.P., India

    Abhishek Walia, Anjali Chauhan & C. K. Shirkot

  2. Department of Biotechnology, National Institute of Plant Genome Research, Govt. of India, New Delhi, 110067, India

    Preeti Mehta

  3. School of Biotechnology, Shoolini University of Biotechnology and Management Sciences, Solan, 173212, H.P., India

    Saurabh Kulshrestha

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  1. Abhishek Walia
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Correspondence to C. K. Shirkot.

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Walia, A., Mehta, P., Chauhan, A. et al. Purification and characterization of cellulase-free low molecular weight endo β-1,4 xylanase from an alkalophilic Cellulosimicrobium cellulans CKMX1 isolated from mushroom compost. World J Microbiol Biotechnol 30, 2597–2608 (2014). https://doi.org/10.1007/s11274-014-1683-3

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