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Purification and characterisation of two extremely halotolerant xylanases from a novel halophilic bacterium

Abstract

The present work reports for the first time the purification and characterisation of two extremely halotolerant endo-xylanases from a novel halophilic bacterium, strain CL8. Purification of the two xylanases, Xyl 1 and 2, was achieved by anion exchange and hydrophobic interaction chromatography. The enzymes had relative molecular masses of 43 kDa and 62 kDa and pI of 5.0 and 3.4 respectively. Stimulation of activity by Ca2+, Mn2+, Mg2+, Ba2+, Li2+, NaN3 and isopropanol was observed. The K m and V max values determined for Xyl 1 with 4-O-methyl-d-glucuronoxylan are 5 mg/ml and 125,000 nkat/mg respectively. The corresponding values for Xyl 2 were 1 mg/ml and 143,000 nkat/mg protein. Xylobiose and xylotriose were the major end products for both endoxylanases. The xylanases were stable at pH 4–11 showing pH optima around pH 6. Xyl 1 shows maximal activity at 60°C, Xyl 2 at 65°C (at 4 M NaCl). The xylanases showed high temperature stability with half-lives at 60°C of 97 min and 192 min respectively. Both xylanases showed optimal activity at 1 M NaCl, but substantial activity remained for both enzymes at 5 M NaCl.

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Acknowledgments

We thank Dr. Jürgen Puls for generous help with product analysis and Jens Frisbæk Sørensen at Danisco A/S for performing the isoelectric focusing analysis. Rene Trinderup is also acknowledged of his information on the characteristics of strain CL8.

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Correspondence to Kjeld Ingvorsen.

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Communicated by W.D. Grant

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Wejse, P.L., Ingvorsen, K. & Mortensen, K.K. Purification and characterisation of two extremely halotolerant xylanases from a novel halophilic bacterium. Extremophiles 7, 423–431 (2003). https://doi.org/10.1007/s00792-003-0342-7

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Keywords

  • Endo-xylanase
  • Enzyme purification
  • Halophilic
  • Halotolerance
  • Transglycosidation