Applied Biochemistry and Biotechnology

, Volume 173, Issue 8, pp 2099–2115 | Cite as

Production of Cellobiose Dehydrogenase from a Newly Isolated White Rot Fungus Termitomyces sp. OE147

  • Gupteshwar Gupta
  • Rishabh Gangwar
  • Ashwani Gautam
  • Lalit Kumar
  • Anuj Dhariwal
  • Vikram Sahai
  • Saroj Mishra
Article
First Online:
Received:
Accepted:

Abstract

Class I cellobiose dehydrogenases (CDHs) are extracellular hemoflavo enzymes produced at low levels by the Basidiomycetes (white rot fungi). In presence of suitable electron acceptors, e.g., cytochrome c, 2,6-dichlorophenol-indophenol, or metal ions, it oxidizes cellobiose to cellobionolactone. A stringent requirement for disaccharides makes CDH also useful for conversion of lactose to lactobionic acid, an important ingredient in pharma and detergent industry. In this work, class I CDH was produced using a newly identified white rot fungus Termitomyces sp. OE147. Four media were evaluated for CDH production, and maximum enzyme activity of 0.92 international unit (IU)/ml was obtained on Ludwig medium under submerged conditions. Statistical optimization of N source, which had significant effect on CDH production, using Box–Behnken design followed by optimization of inoculum size and age resulted in an increase in activity to 2.9 IU/ml and a productivity of ~25 IU/l/h. The nearly purified CDH exhibited high activity of 26.4 IU/mg protein on lactose indicating this enzyme to be useful for lactobionic acid synthesis. Some of the internal peptide sequences bore 100 % homology to the CDH produced in Myceliophthora thermophila. The fungal isolate was amenable to scale up, and an overall productivity of ~18 IU/l/h was obtained at 14-l level.

Keywords

Termitomyces sp. OE 147 Cellobiose dehydrogenase Lactobionic acid Optimized Ludwig medium 
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Notes

Acknowledgments

Authors gratefully acknowledge the financial assistance received from the Department of Biotechnology (Government of India) for carrying out this study. Further, we are thankful to Dr. R. C. Upadhyay from the National Research Centre for Mushroom (NRCM) Solan, India, for providing the cultures. The authors would also like to acknowledge the help of Mr. M.V.R.K. Sarma for the analysis of the statistical data.

Supplementary material

12010_2014_1010_MOESM1_ESM.doc (52 kb)
Supplementary Fig. 1 (DOC 52 kb)
12010_2014_1010_MOESM2_ESM.doc (37 kb)
Supplementary Fig. 2 (DOC 37 kb)

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Gupteshwar Gupta
    • 1
  • Rishabh Gangwar
    • 1
  • Ashwani Gautam
    • 1
  • Lalit Kumar
    • 1
  • Anuj Dhariwal
    • 2
  • Vikram Sahai
    • 1
  • Saroj Mishra
    • 1
  1. 1.Department of Biochemical Engineering and BiotechnologyIndian Institute of Technology DelhiNew DelhiIndia
  2. 2.JAIV Technologies Pvt., Ltd.New DelhiIndia

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