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

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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).

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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.

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