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Biochemical and Thermodynamic Characterization of a Novel, Low Molecular Weight Xylanase from Bacillus Methylotrophicus CSB40 Isolated from Traditional Korean Food

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Abstract

A low molecular weight xylanase from Bacillus strain CSB40, isolated from traditional Korean food and produced in beechwood xylan, was biochemically and thermodynamically characterized. It was purified 8.12-fold with a 15.88 % yield using DEAE sepharose fast flow, and it was determined to have a mass of ∼27 kDa via sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and xylan zymography. The purified xylanase was optimally active at 50 °C and pH 6 and stable over a wide range of pH (4.5–12.5). The N-terminal amino acid sequence of xylanase was GIQQGDDGKL. The activation energy for beechwood xylan hydrolysis was 29.39 kJmol−1 with k cat value of 927.582 × 102 s−1. K m and V max were 0.080 mg/ml and 794.63 mmol min−1 mg−1. The analysis of other thermodynamic parameters like ∆H, ∆G, ∆S, Q10, ∆GE-S, and ∆GE-T also supported the spontaneous formation of products, greater hydrolytic efficiency, and feasibility of enzymatic reaction, which also ratifies the novelty of this xylanase. The enzyme was strongly activated by Zn2+ and inhibited by Cu2+. The principal hydrolyzed end-products of this xylanase are xylobiose, xylotriose, and xylotetrose, which can be used in the pharmaceutical industry and as prebiotic in food.

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Acknowledgments

This work was supported by research fund from Chosun University, 2015.

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    Authors and Affiliations

    1. Department of Pharmacy, Chosun University, Gwangju, Korea

      Sandesh Panthi, Yoon Seok Choi, Yun Hee Choi, MiRi Kim & Jin Cheol Yoo

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    1. Sandesh Panthi
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    2. Yoon Seok Choi
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    3. Yun Hee Choi
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    Correspondence to Jin Cheol Yoo.

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    Sandesh Panthi and Yoon Seok Choi contributed equally to this work.

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    Panthi, S., Choi, Y.S., Choi, Y.H. et al. Biochemical and Thermodynamic Characterization of a Novel, Low Molecular Weight Xylanase from Bacillus Methylotrophicus CSB40 Isolated from Traditional Korean Food. Appl Biochem Biotechnol 179, 126–142 (2016). https://doi.org/10.1007/s12010-016-1983-1

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