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Characterization and Application of a Robust Glucose Dehydrogenase from Paenibacillus pini for Cofactor Regeneration in Biocatalysis

  • Shikha Shah
  • Avinash Vellore Sunder
  • Pooja Singh
  • Pramod P. WangikarEmail author
Original research article
  • 20 Downloads

Abstract

Glucose dehydrogenases are important auxiliary enzymes in biocatalysis, employed in the regeneration of reduced nicotinamide cofactors for oxidoreductase catalysed reactions. Here we report the identification and characterization of a novel glucose-1-dehydrogenase (GDH) from Paenibacillus pini that prefers NAD+ as cofactor over NADP+. The purified recombinant P. pini GDH displayed a specific activity of 247.5 U/mg. The enzyme was stable in the pH range 4–8.5 and exhibited excellent thermostability till 50 °C for 24 h, even in the absence of NaCl or glycerol. Paenibacillus pini GDH was also tolerant to organic solvents, demonstrating its potential for recycling cofactors for biotransformation. The potential application of the enzyme was evaluated by coupling with a NAD+-dependent alcohol dehydrogenase for the reduction of acetophenone and ethyl-4-chloro-3-oxo-butanoate. Conversions higher than 95% were achieved within 2 h with low enzyme loading using lyophilized cell lysate, suggesting that P. pini GDH could be highly effective for recycling NADH in redox biocatalysis.

Keywords

Cofactor regeneration Oxidoreductase Dehydrogenase Biocatalysis Paenibacillus pini 

Notes

Acknowledgements

AVS acknowledges the award of National Postdoctoral Fellowship by the Science and Engineering Research Board (SERB) (Grant No. PDF_2016_003685), Government of India. The authors acknowledge financial support from the Wadhwani Research Center for Bioengineering at Indian Institute of Technology Bombay.

Author’s Contribution

AVS and PW designed the research. SS, PS and AVS performed the research and analysed the data. SS, AVS and PW wrote the paper. All authors read and approved the final manuscript.

Supplementary material

12088_2019_834_MOESM1_ESM.docx (509 kb)
Supplementary material 1 (DOCX 508 kb)

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Copyright information

© Association of Microbiologists of India 2019

Authors and Affiliations

  • Shikha Shah
    • 1
  • Avinash Vellore Sunder
    • 1
  • Pooja Singh
    • 1
    • 2
  • Pramod P. Wangikar
    • 1
    Email author
  1. 1.Department of Chemical EngineeringIndian Institute of Technology BombayPowai, MumbaiIndia
  2. 2.Department of BiochemistrySavitribai Phule Pune UniversityPuneIndia

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