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Cloning and characterization of a Flavin-free oxygen-insensitive azoreductase from Klebsiella oxytoca GS-4-08

  • Jing-qiu Hua
  • Lei YuEmail author
Original Research Paper

Abstract

Objectives

This study reports the identification of a new bacterial azoreductase (AzoR) from Klebsiella oxytoca GS-4-08, its heterologous production in Escherichia coli and the decolorization of azo dyes.

Results

The AzoR, as a flavin-free, oxygen-insensitive enzyme, has a molecular mass of 22 kDa and a high substrate specificity for methyl red (MR). In the presence of 5 mM of NADH, the enzyme activity for decolorization of 50 mg l−1 of MR reached 5.61 μmol min−1 mg protein−1. Double-reciprocal plots indicated that the NADH and MR reductions proceed by ping-pong mechanism. The calculated Vm was 8.17 μM min−1 mg protein−1, and the AzoR retained over 75% of relative activity under temperature range from 20 to 50 °C.

Conclusion

This study for the first time identified an AzoR from Klebsiella oxytoca strain, which enabled efficient degradation of azo dyes by strain GS-4-08.

Keywords

Azoreductase Flavin Klebsiella oxytoca Oxygen-insensitive 

Notes

Acknowledgements

This research was supported by the National Natural Science Foundation of China (No. 51778298) and the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD).

Compliance with ethical standards

Conflict of interest

The Authors declare that there is no conflict of interest.

Supplementary material

10529_2019_2647_MOESM1_ESM.docx (298 kb)
Supplementary material 1 (DOCX 297 kb)

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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Department of Environmental EngineeringNanjing Forestry UniversityNanjingChina
  2. 2.College of Biology and the Environment, Co-Innovation Center for Sustainable Forestry in Southern ChinaNanjing Forestry UniversityNanjingChina

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