Purification and Characterization of Cellobiose Dehydrogenase, a Novel Extracellular Hemoflavoenzyme from the White-Rot Fungus Phanerochaete chrysosporium

Bao, Wenjun; Usha, S. N.; Renganathan, V.

doi:10.1006/abbi.1993.1098

Cited by 143 publications

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“…9) have not yielded positive results. An enzymatic precedent for the oxidation of the anomeric carbon at the reducing end of an oligosaccharide has been reported in the case of cellobiose dehydrogenase (30).…”

Section: Fig 6 Subcellular Localization Of Enzymes That Act On (Kdomentioning

confidence: 99%

Lipopolysaccharide Biosynthesis in Rhizobium leguminosarum

Brozek

Kadrmas

Raetz

1996

Journal of Biological Chemistry

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The lipopolysaccharide of Rhizobium leguminosarum differs from that of other Gram-negative organisms. R. leguminosarum lipid A lacks phosphate groups, but it contains a galacturonic acid residue at the 4-position and an aminogluconate moiety in place of the usual glucosamine 1-phosphate unit. R. leguminosarum lipid A is esterified with a peculiar long chain fatty acid, 27-hydroxyoctacosanoate, not found in enteric Gramnegative bacteria, and the inner core of R. leguminosarum contains mannose and galactose in place of heptose. Despite these differences, the biosynthesis of R. leguminosarum In addition, at least two hydrophobic metabolites were generated from (Kdo) 2 -[4-32 P]-lipid IV A in a manner that was dependent upon both membranes and a cytosolic factor from R. leguminosarum. These compounds are attributed to novel acylations of (Kdo) 2 -[4-32 P]-lipid IV A . E. coli membranes and cytosol did not catalyze any of the unique reactions detected in R. leguminosarum extracts. Our findings establish the conservation and versatility of (Kdo) 2 -lipid IV A as a lipid A precursor in bacteria.

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Section: Fig 6 Subcellular Localization Of Enzymes That Act On (Kdomentioning

confidence: 99%

Lipopolysaccharide Biosynthesis in Rhizobium leguminosarum

Brozek

Kadrmas

Raetz

1996

Journal of Biological Chemistry

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“…Besides cellobiose, soluble cellodextrins, mannobiose and lactose, are good or acceptable substrates for CDH, whereas monosaccharides are poor substrates (7). CDH can use a large number of electron acceptors; however, reduction of oxygen is slow (11).…”

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confidence: 99%

Mechanism of the Reductive Half-reaction in Cellobiose Dehydrogenase

Hällberg¹,

Henriksson²,

Pettersson³

et al. 2003

Journal of Biological Chemistry

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The extracellular flavocytochrome cellobiose dehydrogenase (CDH; EC 1.1.99.18) participates in lignocellulose degradation by white-rot fungi with a proposed role in the early events of wood degradation. The complete hemoflavoenzyme consists of a catalytically active dehydrogenase fragment (DH cdh ) connected to a b-type cytochrome domain via a linker peptide. In the reductive half-reaction, DH cdh catalyzes the oxidation of cellobiose to yield cellobiono-1,5-lactone. The active site of DH cdh is structurally similar to that of glucose oxidase and cholesterol oxidase, with a conserved histidine residue positioned at the re face of the flavin ring close to the N5 atom. The mechanisms of oxidation in glucose oxidase and cholesterol oxidase are still poorly understood, partly because of lack of experimental structure data or difficulties in interpreting existing data for enzyme-ligand complexes. Here we report the crystal structure of the Phanerochaete chrysosporium DH cdh with a bound inhibitor, cellobiono-1,5-lactam, at 1.8-Å resolution. The distance between the lactam C1 and the flavin N5 is only 2.9 Å, implying that in an approximately planar transition state, the maximum distance for the axial 1-hydrogen to travel for covalent addition to N5 is 0.8 -0.9 Å. The lactam O1 interacts intimately with the side chains of His-689 and Asn-732. Our data lend substantial structural support to a reaction mechanism where His-689 acts as a general base by abstracting the O1 hydroxyl proton in concert with transfer of the C1 hydrogen as hydride to the re face of the flavin N5.Cellobiose dehydrogenases (CDHs 1 ; EC 1.1.99.18) are extracellular fungal flavocytochromes that are believed to participate in lignocellulose degradation by fungi. They are oxidoreductases carrying protoheme and FAD cofactors bound to separate domains. In vitro, CDH from the white-rot Basidiomycete Phanerochaete chrysosporium depolymerizes cellulose, hemicelluloses, and lignin (Refs. 1-3; for review, see Ref. 4) as well as other polymers (5). The exact biological function of CDH has been a subject of lively debate, but recent results suggest that the enzyme is important for invasion and colonization of wood (6).The catalytic site is located in the flavoprotein domain, where the reductive half-reaction proceeds by oxidation of ␤-cellobiose (apparent k cat 15.7 s Ϫ1 and K m 0.11 mM, see Ref. 7) to yield cellobiono-1,5-lactone (Fig.

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“…Cellobiose dehydrogenase has been isolated and characterized from a variety of white-rot fungi (2,6,21,38,43,53), soft-rot fungi (13,15,20,50,52), and one brown-rot fungus (49). Typically, CDH is a monomeric protein consisting of two domains, one containing a hexacoordinate heme b (14) and one containing a noncovalently bound flavin adenine dinucleotide (FAD) (6,43) or 6-hydroxy FAD (33).…”

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Purification and Characterization of Cellobiose Dehydrogenase from the Plant PathogenSclerotium(Athelia)rolfsii

Baminger

Subramaniam

Renganathan

et al. 2001

Appl Environ Microbiol

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Cellobiose dehydrogenase (CDH) is an extracellular hemoflavoenzyme produced by several wood-degrading fungi. In the presence of a suitable electron acceptor, e.g., 2,6-dichloro-indophenol (DCIP), cytochrome c, or metal ions, CDH oxidizes cellobiose to cellobionolactone. The phytopathogenic fungus Sclerotium rolfsii (teleomorph: Athelia rolfsii) strain CBS 191.62 produces remarkably high levels of CDH activity when grown on a cellulose-containing medium. Of the 7,500 U of extracellular enzyme activity formed per liter, less than 10% can be attributed to the proteolytic product cellobiose:quinone oxidoreductase. As with CDH from wood-rotting fungi, the intact, monomeric enzyme from S. rolfsii contains one heme b and one flavin adenine dinucleotide cofactor per molecule. It has a molecular size of 101 kDa, of which 15% is glycosylation, and a pI value of 4.2. The preferred substrates are cellobiose and cellooligosaccharides; additionally, ␤-lactose, thiocellobiose, and xylobiose are efficiently oxidized. Cytochrome c (equine) and the azino-di-(3-ethyl-benzthiazolin-6-sulfonic acid) cation radical were the best electron acceptors, while DCIP, 1,4-benzoquinone, phenothiazine dyes such as methylene blue, phenoxazine dyes such as Meldola's blue, and ferricyanide were also excellent acceptors. In addition, electrons can be transferred to oxygen. Limited in vitro proteolysis with papain resulted in the formation of several protein fragments that are active with DCIP but not with cytochrome c. Such a flavin-containing fragment, with a mass of 75 kDa and a pI of 5.1 and lacking the heme domain, was isolated and partially characterized.

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Purification and Characterization of Cellobiose Dehydrogenase, a Novel Extracellular Hemoflavoenzyme from the White-Rot Fungus Phanerochaete chrysosporium

Cited by 143 publications

References 0 publications

Lipopolysaccharide Biosynthesis in Rhizobium leguminosarum

Lipopolysaccharide Biosynthesis in Rhizobium leguminosarum

Mechanism of the Reductive Half-reaction in Cellobiose Dehydrogenase

Purification and Characterization of Cellobiose Dehydrogenase from the Plant PathogenSclerotium(Athelia)rolfsii

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