The genes for three xylan-degrading activities from Bacteroides ovatus are clustered in a 3.8-kilobase region.
Genes coding for three xylan-degrading activities, xylanase, xylosidase, and arabinosidase, were simultaneously cloned from the colonic anaerobic organism Bacteriodes ovatus. The genes for the three enzymes were located on a 3.8-kilobase EcoRI genomic insert and were cloned by using plasmid pUC18. All three activities were expressed in Escherichia coli JM83, and all were cell associated. Expression of the xylanase gene was independent from expression of the xylosidase and arabinosidase genes, whereas expression of the latter two genes appeared to be coordinated. Restriction endonuclease analysis of the arabinosidase and xylosidase genes and partial purification of these enzyme activities from E. coli suggested that these activities were catalyzed by a bifunctional protein or two proteins of very similar molecular weight. All three enzyme activities were regulated in B. ovatus in response to the carbon source used for growth. This is the first report of the cloning and expression of B. ovatus genes.
Whitehead
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Northern Regional Research Center, Department of Agriculture, Peoria, Illinois 61604.
Hespell
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D001439:Bacteroides / Q000201:enzymology / Q000235:genetics*; D002855:Chromatography, Thin Layer; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004790:Enzyme Induction; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D006026:Glycoside Hydrolases / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D008970:Molecular Weight; D005810:Multigene Family; D009844:Oligosaccharides / Q000302:isolation & purification; D010957:Plasmids; D011134:Polysaccharides / Q000378:metabolism*; D015183:Restriction Mapping; D013379:Substrate Specificity; D043325:Xylan Endo-1,3-beta-Xylosidase; D014990:Xylans / Q000378:metabolism*; D014995:Xylosidases / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism
D009844:Oligosaccharides; D011134:Polysaccharides; D014990:Xylans; D006026:Glycoside Hydrolases; D014995:Xylosidases; D043325:Xylan Endo-1,3-beta-Xylosidase; C019713:alpha-N-arabinofuranosidase