Expression, Purification, and Characterization of the Recombinant Putative Periplasmic Hemin-Binding Protein (HutB) of Photobacterium damselae subsp. piscicida

Francesca ANDREONI; Romina BOIANI; Giordano SERAFINI; Irene BIANCONI; Sabrina DOMINICI; Francesca GORINI; Mauro MAGNANI

doi:10.1271/bbb.80732

Biochemistry & Molecular Biology Notes

Expression, Purification, and Characterization of the Recombinant Putative Periplasmic Hemin-Binding Protein (HutB) of Photobacterium damselae subsp. piscicida

Francesca ANDREONI, Romina BOIANI, Giordano SERAFINI, Irene BIANCONI, Sabrina DOMINICI, Francesca GORINI, Mauro MAGNANI

Author information

Francesca ANDREONI
Department of Biomolecular Science, Section of Biotechnology, University of Urbino “Carlo Bo,” via T. Campanella
Romina BOIANI
Department of Biomolecular Science, Section of Biotechnology, University of Urbino “Carlo Bo,” via T. Campanella
Giordano SERAFINI
Department of Biomolecular Science, Section of Biochemistry and Molecular Biology, University of Urbino “Carlo Bo,” via Saffi
Irene BIANCONI
Department of Biomolecular Science, Section of Biotechnology, University of Urbino “Carlo Bo,” via T. Campanella
Sabrina DOMINICI
Department of Biomolecular Science, Section of Biochemistry and Molecular Biology, University of Urbino “Carlo Bo,” via Saffi
Francesca GORINI
Department of Biomolecular Science, Section of Biotechnology, University of Urbino “Carlo Bo,” via T. Campanella
Mauro MAGNANI
Department of Biomolecular Science, Section of Biotechnology, University of Urbino “Carlo Bo,” via T. Campanella
Department of Biomolecular Science, Section of Biochemistry and Molecular Biology, University of Urbino “Carlo Bo,” via Saffi

Keywords: Photobacterium damselae, periplasmic hemin-binding protein (HutB), hemin binding protein, hemin, recombinant protein

JOURNAL FREE ACCESS

2009 Volume 73 Issue 5 Pages 1180-1183

DOI https://doi.org/10.1271/bbb.80732

View "Advance Publication" version (May 7, 2009).

Details

Abstract

HutB, the periplasmic hemin binding protein of Photobacterium damselae subsp. piscicida, was produced as a recombinant protein. UV-Vis spectrophotometrical analysis showed absorption spectral changes in hemin upon mixing it with the recombinant protein, indicating complex formation. Spectrophotometric titration of HutB with hemin showed saturation at a heme/HutB ratio of 1:1 and a binding affinity (K_d) of 10 μM.

Corresponding author

Register with J-STAGE for free!