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Biotechnology Letters

, Volume 32, Issue 7, pp 921–927 | Cite as

Site-directed mutagenesis enhances the activity of NADH-FMN oxidoreductase (DszD) activity of Rhodococcus erythropolis

  • Nasrin Kamali
  • Mahmood Tavallaie
  • Bijan Bambai
  • Ali Asghar Karkhane
  • Mandana Miri
Original Research Paper

Abstract

Microbial desulfurization is potentially an alternative process to chemical desulfurization of fossil fuels and their refined products. The dibenzothiophene desulfurizing system of Rhodococcus erythropolis includes DszD which is an NADH-dependent FMN oxidoreductase with 192 residues that is responsible for supplying reducing equivalents in the form of FMNH2 to monooxygenases, DszA and DszC. We performed amino acid sequence comparisons and structural predictions based on the crystal structure of available pdb files for three flavin reductases PheA2, HpaCTt and HpaCSt with the closest structural homology to IGTS8 DszD. The Thr62 residue in DszD was substituted with Asn and Ala by site-directed single amino acid mutagenesis. Variants T62N and T62A showed 5 and 7 fold increase in activities based on the recombinant wild type DszD, respectively. This study revealed the critical role of position 62 in enzyme activity. These results represent the first experimental report on flavin reductase mutation in R. erythropolis and will pave the way for further optimization of the biodesulfurization process.

Keywords

Biodesulfurization DszD FMN oxidoreductase Rhodococcus erythropolis IGTS8 Site-directed mutagenesis 

Notes

Acknowledgements

This work was supported by a research grant from the Malek Ashtar Industrial University. Authors appreciate the technical assistance of S. Zeidabadi and acknowledge Dr. P. Shariati for reading this manuscript.

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Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Nasrin Kamali
    • 1
  • Mahmood Tavallaie
    • 2
  • Bijan Bambai
    • 1
    • 4
  • Ali Asghar Karkhane
    • 1
  • Mandana Miri
    • 3
  1. 1.National Institute for Genetic Engineering and Biotechnology (NIGEB)TehranIran
  2. 2.Bagyatallah Medical Sciences UniversityHuman Genetic Research CenterTehranIran
  3. 3.Science and Research Branch of TehranIslamic Azad UniversityTehranIran
  4. 4.Faculty of Biological SciencesShahid Beheshti UniversityTehranIran

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