Heterologous expression of the Bacteroides ruminicola xylanase gene in Bacteroides fragilis and Bacteroides uniformis.
A cloned xylanase gene from the ruminal bacterium Bacteroides ruminicola 23 was transferred by conjugation into the colonic species Bacteroides fragilis and Bacteroides uniformis by using the Escherichia coli-Bacteroides shuttle vector pVAL-1. The cloned gene was expressed in both species, and xylanase specific activity in crude extracts was found to be at least 1400-fold greater than that found in the B. ruminicola strain. Analysis of crude extract proteins from the recombinant B. fragilis by SDS-PAGE demonstrated a new 60,000 molecular weight protein. The xylanase activity expressed in both E. coli and B. fragilis was capable of degrading xylan to xylooligosaccharides in vitro. This is the first demonstration that colonic Bacteroides species can express a gene from a ruminal Bacteroides species.
Whitehead
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Agricultural Research Service, U.S. Department of Agriculture, Peoria, IL 61604.
Hespell
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D001439:Bacteroides / Q000201:enzymology / Q000235:genetics*; D001441:Bacteroides fragilis / Q000201:enzymology / Q000235:genetics; D003001:Cloning, Molecular; D003227:Conjugation, Genetic; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D005798:Genes, Bacterial; D006026:Glycoside Hydrolases / Q000235:genetics*; D010957:Plasmids; D013045:Species Specificity; D043325:Xylan Endo-1,3-beta-Xylosidase
D006026:Glycoside Hydrolases; D043325:Xylan Endo-1,3-beta-Xylosidase