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Bioprocess and Biosystems Engineering

, Volume 40, Issue 5, pp 651–662 | Cite as

Production and characteristics of the recombinant extracellular bifunctional endoglucanase of the polyextremophilic bacterium Bacillus halodurans and its applicability in saccharifying agro-residues

  • Ranjitha R. Prabhu
  • Deepak Parashar
  • T. SatyanarayanaEmail author
Research Paper

Abstract

The recombinant alkalistable and moderately thermostable bifunctional endoglucanase gene (BhCell-Xyl) of polyextremophilic bacterium Bacillus halodurans TSLV1 has been expressed in Pichia pastoris under constitutive GAP as well as inducible AOX promoters. A higher titre of recombinant BhCell-Xyl was attained after induction (4.8 U mL−1) as compared to that of the constitutive production (2.1 U mL−1). The recombinant P. pastoris strains integrated two copies of BhCell-Xyl under AOX and GAP promoters. The pure recombinant BhCell-Xyl is a glycoprotein of 66 kDa, which is optimally active at 60 °C and pH 6.0 and 8.0. Glycosylated BhCell-Xyl exhibits higher thermostability than that of the native enzyme. The analysis of amino acids of BhCell-Xyl revealed that multiple factors are responsible for its thermostability. Kinetics and in silico analysis of the enzyme suggested that BhCell-Xyl has one active site for both endocellulase and endoxylanase activities. The BhCell-Xyl possesses a carbohydrate binding domain and saccharifies lignocellulosic agro-residues to xylo-oligosaccharides and cello-oligosaccharides, suggesting its potential application in generating fermentable sugars from renewable agro-residues for biofuel and fine chemical industries.

Keywords

Bacillus halodurans Bifunctional enzyme Endoglucanase P. pastoris Polyextremophilic 

Notes

Acknowledgements

Authors gratefully acknowledge financial assistance from the Indo-US Science and Technology Forum and Department of Biotechnology, Govt. of India, New Delhi and University Grants Commission, New Delhi while carrying out the work presented in this manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interests.

Supplementary material

449_2016_1730_MOESM1_ESM.pdf (294 kb)
Supplementary material 1 (PDF 294 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Ranjitha R. Prabhu
    • 1
  • Deepak Parashar
    • 1
  • T. Satyanarayana
    • 1
    Email author
  1. 1.Department of MicrobiologyUniversity of Delhi South CampusNew DelhiIndia

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