Abstract
The cel1 gene of Agaricus bisporus encodes a protein (CEL1) that has an architecture resembling the multi-domain fungal cellulases, although the sequence of its putative catalytic core is not matched by any other in the protein and nucleic acid data bases. The N-terminal half of the putative catalytic domain of CEL1 was expressed in Escherichia coli as a fusion protein with glutathione-S-transferase. The fusion protein was used to raise a CEL1-specific antibody. CEL1 was detected as an extracellular 49.8 kDa protein in A. bisporus cellulose-grown cultures, where it bound strongly to cellulose. CEL1 was neither an endoglucanase, a cellobiohydrolase able to hydrolyze fluorogenic cellobiosides, a beta-glucosidase, a xylanase, nor a cellobiose: quinone oxidoreductase. CEL1 was present in some fractions of culture fluid separated by electrophoresis which released soluble sugars from crystalline cellulose.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Agaricus / genetics
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Agaricus / growth & development
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Agaricus / metabolism*
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Amino Acid Sequence
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Antibodies, Fungal / biosynthesis
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Base Sequence
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Carrier Proteins / biosynthesis
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Carrier Proteins / genetics
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Carrier Proteins / immunology
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Cellulase / biosynthesis*
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Cellulase / genetics
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Cellulase / immunology
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Cellulose / metabolism*
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Cellulose 1,4-beta-Cellobiosidase
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Cloning, Molecular
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DNA, Fungal / genetics
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Escherichia coli / genetics
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Fungal Proteins / biosynthesis*
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Fungal Proteins / genetics
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Fungal Proteins / immunology
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Genes, Fungal
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Genetic Vectors
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Glutathione Transferase / biosynthesis
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Glutathione Transferase / genetics
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Molecular Sequence Data
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Recombinant Fusion Proteins / biosynthesis
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / immunology
Substances
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Antibodies, Fungal
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Carrier Proteins
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DNA, Fungal
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Fungal Proteins
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Recombinant Fusion Proteins
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Cellulose
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Glutathione Transferase
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Cellulase
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Cellulose 1,4-beta-Cellobiosidase