Resonance Raman spectroscopic studies of cellobiose dehydrogenase from Phanerochaete chrysosporium

Archives of Biochemistry and Biophysics
J D CohenT M Loehr

Abstract

Cellobiose dehydrogenase (CDH), an extracellular hemoflavoenzyme produced by cellulose-degrading cultures of Phanerochaete chrysosporium, oxidizes cellobiose to cellobionolactone. The enzyme contains one 6-coordinate, low-spin b-type heme and one FAD cofactor per monomeric protein. In this work, resonance Raman (RR) spectra are reported for the oxidized, reduced, and deflavo forms of CDH as well as the individual flavin and heme domains of the enzyme obtained by peptide proteolysis. The RR spectra of the flavin and heme groups of CDH were assigned by comparison to the spectra of other hemoflavoenzymes and model compounds. Proteolytic cleavage of the CDH domains had only a minimal spectroscopic effect on the vibrational modes of the heme and FAD cofactors. Excitation of the oxidized CDH holoenzyme at 413 or 442 nm resulted in photoreduction of the heme. However, the same excitation wavelength used on the deflavo form of the enzyme or on the heme domain alone did not cause photoreduction, indicating that photoinitiated electron transfer requires the FAD cofactor. These observations suggest an enzymatic mechanism whereby reducing equivalents obtained from the oxidation of cellobiose are transferred from the FAD to the heme. A simi...Continue Reading

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Citations

May 8, 2004·The Journal of Biological Chemistry·Hao ZhuH Franklin Bunn
Aug 9, 2005·Applied and Environmental Microbiology·Makoto YoshidaMasahiro Samejima
Feb 13, 2001·Enzyme and Microbial Technology·M D. Cameron, S D. Aust
Jul 28, 2010·Chemphyschem : a European Journal of Chemical Physics and Physical Chemistry·Roland LudwigLo Gorton
Dec 21, 2002·The Journal of Biological Chemistry·B Martin HallbergChristina Divne
Jun 11, 2003·The Journal of Biological Chemistry·Frederik A J RotsaertMichael H Gold
Apr 25, 2015·Chemphyschem : a European Journal of Chemical Physics and Physical Chemistry·Patrycja KielbInez M Weidinger
Nov 9, 2019·ChemElectroChem·Marta MeneghelloPhilip N Bartlett
Mar 22, 2000·Journal of Biotechnology·G HenrikssonG Pettersson
Jan 12, 2002·Journal of Molecular Biology·B M HallbergChristina Divne

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