Abstract
Cellobiose dehydrogenase (CDH; EC 1.1.99.18) is an extracellular glycosylated protein composed of two distinct domains, a C-terminal catalytic flavin domain and an N-terminal cytochrome-b-type heme domain, which transfers electrons from the flavin domain to external electron acceptors. The soluble flavin domain of the Phanerochaete chrysosporium CDH was successfully expressed in Escherichia coli. The enzyme showed dye-mediated CDH activity higher than that of the complete CDH, composed of flavin domain and heme domain, prepared using Pichia pastoris as the host microorganism. The ability to conveniently express the recombinant CDH flavin domain in E. coli provides great opportunities for the molecular engineering of the catalytic properties of CDH.
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References
Cameron MD, Aust SD (2001) Cellobiose dehydrogenase an extracellular fungal flavocytochrome. Enzym Microb Technol 28:129–138
Desriani, Ferri S, Sode K (2010) Amino acid substitution at the substrate binding subsite alters the specificity of the Phanerochaete chrysosporium cellobiose dehydrogenase. Biochem Biophy Res Commun (in press)
Hallberg BM, Henriksson G, Pettersson G, Divne C (2002) Crystal structure of the flavoprotein domain of the extracellular flavocytochrome cellobiose dehydrogenase. J Mol Biol 315:421–434
Henriksson G, Pettersson G, Johansson G, Ruiz A, Uzcategui E (1991) Cellobiose oxidase from Phanerochaete chrysosporium can be cleaved by papain into two domains. Eur J Biochem 196:101–106
Henriksson G, Sild V, Szabo IJ, Pettersson G, Johansson G (1998) Substrate specificity of cellobiose dehydrogenase from Phanerochaete chrysosporium. Biochim Biophys Acta 1383:48–54
Hilden L, Eng L, Johansson G, Lindqvist SE, Pettersson G (2001) An amperometric cellobiose dehydrogenase based biosensor can be used for measurement of cellulose activity. Anal Biochem 290:245–250
Larsson T, Lindgren A, Ruzgas T, Linquist SE, Gorton L (2000) Bioelectrochemical characterization of cellobiose dehydrogenase modified graphite electrodes: ionic strength and pH dependences. J Electroanal Chem 482:1–10
Nikilla H, Gennis RB, Sligar SG (1991) Cloning and expression of the gene encoding the soluble cytochrome b562 of Escherichia coli. Eur J Biochem 202:309–313
Rostaert FAJ, Renganathan V, Gold MH (2003) Role of the flavin domain residues, his689 and asn732, in the catalytic mechanism of cellobiose dehydrogenase from Phanerochaete chrysosporium. Biochemistry 42:4049–4056
Sargent F (2007) The twin-arginine transport system: moving folded proteins across membranes. Biochem Soc Trans 35:835–847
Studier FW (2005) Protein production by auto-induction in high density shaking cultures. Protein Expr Purif 41:207–234
Tasca F, Gorton L, Harreither W, Haltrich D, Ludwig R, Noll G (2009) Comparison of direct and mediated electron transfer for cellobiose dehydrogenase from Phanerochaete chrysosporium. Anal Chem 81:2791–2798
Yoshida M, Ohira T, Igarashi K, Nagasawa H, Aida K, Hallberg BM, Divne C, Nishino T, Samejima M (2001) Production and characterization of recombinant Phanerochaete chrysosporium cellobiose dehydrogenase in the methylotrophic yeast Pichia pastoris. Biosci Biotechnol Biochem 65:2050–2057
Zamocky M, Hallber M, Ludwig R, Divne C, Haltrich D (2004) Ancestral gene fusion in cellobiose dehydrogenase a specific evolution of GMC oxidoreductase in fungi. Gene 338:1–14
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Desriani, Ferri, S. & Sode, K. Functional expression of Phanerochaete chrysosporium cellobiose dehydrogenase flavin domain in Escherichia coli . Biotechnol Lett 32, 855–859 (2010). https://doi.org/10.1007/s10529-010-0215-y
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DOI: https://doi.org/10.1007/s10529-010-0215-y