Abstract
To engineer a multifunctional xylan-degrading enzyme, a chimera was created by fusing the xylanase domain of the Clostridium thermocellum xylanase (xynZ) and a dual functional arabinofuranosidase/xylosidase (DeAFc; from a compost starter mixture) through a flexible peptide linker. The xylanase domain of xynZ possesses previously unreported endoglucanase activity. The chimera, possessing the activities of xylanase, endoglucanase, arabinofuranosidase and xylosidase, was expressed in E. coli and purified. The chimera closely resembled the parental enzymes in pH, temperature optima and kinetics, and was more active than the parental enzyme mixture in the hydrolysis of natural xylans and corn stover.
Publication types
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Research Support, U.S. Gov't, Non-P.H.S.
MeSH terms
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Clostridium thermocellum / enzymology
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Clostridium thermocellum / genetics
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Endo-1,4-beta Xylanases / genetics
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Endo-1,4-beta Xylanases / metabolism
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Enzyme Stability
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Escherichia coli / genetics
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Gene Expression
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Glycoside Hydrolases / chemistry
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Glycoside Hydrolases / genetics*
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Glycoside Hydrolases / isolation & purification
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Glycoside Hydrolases / metabolism*
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Hydrogen-Ion Concentration
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Kinetics
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Models, Molecular
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Recombinant Fusion Proteins / chemistry
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / isolation & purification
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Recombinant Fusion Proteins / metabolism
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Temperature
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Xylans / metabolism*
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Xylosidases / genetics
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Xylosidases / metabolism
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Zea mays / metabolism
Substances
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Recombinant Fusion Proteins
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Xylans
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Glycoside Hydrolases
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Xylosidases
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hemicellulase
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alpha-N-arabinofuranosidase
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Endo-1,4-beta Xylanases