Discovery of a Xylooligosaccharide Oxidase from Myceliophthora thermophila C1

By inspection of the predicted proteome of the fungus Myceliophthora thermophila C1 for vanillyl-alcohol oxidase (VAO)-type flavoprotein oxidases, a putative oligosaccharide oxidase was identified. By homologous expression and subsequent purification, the respective protein could be obtained. The pr...

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Bibliographic Details
Published in:The Journal of biological chemistry 2016-11, Vol.291 (45), p.23709-23718
Main Authors: Ferrari, Alessandro R., Rozeboom, Henriëtte J., Dobruchowska, Justyna M., van Leeuwen, Sander S., Vugts, Aniek S.C., Koetsier, Martijn J., Visser, Jaap, Fraaije, Marco W.
Format: Article
Language:English
Subjects:
Amino Acid Sequence
carbohydrate
carbohydrate processing
CAZy
crystal structure
Crystallography, X-Ray
Disaccharides - metabolism
Enzymology
flavin adenine dinucleotide (FAD)
Flavin-Adenine Dinucleotide - metabolism
Glucuronates - metabolism
Models, Molecular
oligosaccharide
Oligosaccharides - metabolism
oxidase
Oxidation-Reduction
Oxidoreductases - chemistry
Oxidoreductases - metabolism
Protein Conformation
Sequence Alignment
Sordariales - chemistry
Sordariales - enzymology
Sordariales - metabolism
Substrate Specificity
xylan
Xylans - metabolism
xylose
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Summary:By inspection of the predicted proteome of the fungus Myceliophthora thermophila C1 for vanillyl-alcohol oxidase (VAO)-type flavoprotein oxidases, a putative oligosaccharide oxidase was identified. By homologous expression and subsequent purification, the respective protein could be obtained. The protein was found to contain a bicovalently bound FAD cofactor. By screening a large number of carbohydrates, several mono- and oligosaccharides could be identified as substrates. The enzyme exhibits a strong substrate preference toward xylooligosaccharides; hence it is named xylooligosaccharide oxidase (XylO). Chemical analyses of the product formed upon oxidation of xylobiose revealed that the oxidation occurs at C1, yielding xylobionate as product. By elucidation of several XylO crystal structures (in complex with a substrate mimic, xylose, and xylobiose), the residues that tune the unique substrate specificity and regioselectivity could be identified. The discovery of this novel oligosaccharide oxidase reveals that the VAO-type flavoprotein family harbors oxidases tuned for specific oligosaccharides. The unique substrate profile of XylO hints at a role in the degradation of xylan-derived oligosaccharides by the fungus M. thermophila C1.
ISSN:0021-9258
1083-351X
1083-351X
DOI:10.1074/jbc.M116.741173