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Expression, purification and characterization of flavin reductase from Citrobacter freundii A1

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Abstract

Flavin reductase plays an important biological role in catalyzing the reduction of flavin by NAD(P)H oxidation. The gene that codes for flavin reductase from Citrobacter freundii A1 was cloned and expressed in Escherichia coli BL21(DE3)pLysS. In this study, we aimed to characterize the purified recombinant flavin reductase of C. freundii A1. The recombinant enzyme was purified to homogeneity and the biochemical profiles, including the effect of pH, temperature, metal ions and anions on flavin reductase activity and stability, were determined. This enzyme exhibited optimum activity at 45 °C in a 10-min reaction at pH 7.5 and was stable at temperatures up to 30 °C. At 0.1 mM concentration of metal ions, flavin reductase activity was stimulated by divalent cations including Mn2+, Sr2+, Ni2+, Sn2+, Ba2+, Co2+, Mg2+, Ca2+ and Pb2+. Ag+ was noticeably the strongest inhibitor of recombinant flavin reductase of C. freundii A1. This enzyme should not be defined as a standard flavoprotein. This is the first attempt to characterize flavin reductase of C. freundii origin.

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Acknowledgments

We would like to acknowledge the Ministry of Science, Technology and Innovation Malaysia (MOSTI) for research grant under Vote No. 74199 and National Science Fellowship (NSF) scholarship awarded to G.F. Chan.

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Authors and Affiliations

  1. Nanoporous Materials for Biological Application Research Group (NAMBAR), Sustainability Research Alliance, Universiti Teknologi Malaysia, 81310 UTM, Johor Bahru, Johor, Malaysia

    Giek Far Chan & Noor Aini Abdul Rashid

  2. Department of Chemistry, Faculty of Science, Universiti Teknologi Malaysia, 81310 UTM, Johor Bahru, Johor, Malaysia

    Abdull Rahim Mohd Yusoff

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  1. Giek Far Chan
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  2. Noor Aini Abdul Rashid
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  3. Abdull Rahim Mohd Yusoff
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Correspondence to Noor Aini Abdul Rashid.

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Chan, G.F., Rashid, N.A.A. & Yusoff, A.R.M. Expression, purification and characterization of flavin reductase from Citrobacter freundii A1. Ann Microbiol 63, 343–351 (2013). https://doi.org/10.1007/s13213-012-0480-1

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  • DOI: https://doi.org/10.1007/s13213-012-0480-1

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