Annals of Microbiology

, Volume 62, Issue 2, pp 825–834 | Cite as

Production, purification and properties of a cellulase-free thermostable endoxylanase from newly isolated Paenibacillus sp. ASCD2

  • Digantkumar Chapla
  • Harshvadan Patel
  • Atmika Singh
  • Datta Madamwar
  • Amita Shah
Original Article

Abstract

A newly isolated thermophillic strain of Paenibacillus sp. ASCD2 was studied for xylanase production under solid state fermentation. The influence of lignocellulosic substrates, moistening media, moisture ratio and pH was analysed; maximum xylanase production (343.52 U/g) was achieved on wheat straw with organic nitrogen rich medium (pH 7.0) at a moisture ratio of 1:5 (w/v) in 72 h at 50°C. After optimization, there was a 3.7-fold increase in xylanase yield. Ammonium sulphate fractionation followed by gel permeation chromatography yielded 16.59-fold purified xylanase with 38.33% recovery. The molecular weight of purified xylanase was found to be 20.2 kDa as estimated by SDS-PAGE. The optimum temperature and pH for xylanase activity of purified xylanase were found to be 60°C and 7.0, respectively. Kinetic parameters Km and Vmax were 2.5 mg/ml and 571 μmol mg−1 min−1, respectively. The enzyme was reasonably thermostable within the temperature range of 55 to 70°C. The half-life of xylanase was 415.12 min and 102.8 min at 65 and 70°C, respectively. Xylanase activity was significantly enhanced by Na+, Mg++ and K+ ions, strongly decreased by EDTA, Hg++, Cu++, Ag++ and Mn++ and unaffected by Fe++, Zn++, and Tween 80. The purified xylanase was found to be an endoxylanase as it produced only xylooligosaccharides from birchwood xylan.

Keywords

Xylanase Thermostable Solid state fermentation Paenibacillus sp. 

Notes

Acknowledgement

The authors gratefully acknowledge the University Grants Commission (UGC), New Delhi, India for providing the research grants to support this work.

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Copyright information

© Springer-Verlag and the University of Milan 2011

Authors and Affiliations

  • Digantkumar Chapla
    • 1
  • Harshvadan Patel
    • 1
  • Atmika Singh
    • 1
  • Datta Madamwar
    • 1
  • Amita Shah
    • 1
  1. 1.BRD School of BiosciencesSardar Patel UniversityVallabh VidyanagarIndia

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