Isolation of genes encoding beta-D-xylanase, beta-D-xylosidase and alpha-L-arabinofuranosidase activities from the rumen bacterium Prevotella ruminicola B1(4)

Prevotella ruminicola B1(4) is a strictly anaerobic, Gram-negative, polysaccharide-degrading rumen bacterium. Xylanase activity in this strain was found to be inducible, the specific activity of cells grown on xylan being increased at least 20-fold by comparison with cells grown on glucose. Ten bact...

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Bibliographic Details
Published in:FEMS microbiology letters 1995-01, Vol.125 (2-3), p.135-141
Main Authors: Gasparic, A, Marinsek-Logar, R, Martin, J, Wallace, R J, Nekrep, F V, Flint, H J
Format: Article
Language:English
Subjects:
Animals
Blotting, Southern
DNA, Bacterial - analysis
DNA, Bacterial - metabolism
Endo-1,4-beta Xylanases
Gene Library
Genes, Fungal
Glycoside Hydrolases - genetics
Glycoside Hydrolases - metabolism
Kinetics
Prevotella - enzymology
Prevotella - genetics
Prevotella - isolation & purification
Recombinant Proteins - metabolism
Restriction Mapping
Rumen - microbiology
Substrate Specificity
Xylosidases - genetics
Xylosidases - metabolism
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Summary:Prevotella ruminicola B1(4) is a strictly anaerobic, Gram-negative, polysaccharide-degrading rumen bacterium. Xylanase activity in this strain was found to be inducible, the specific activity of cells grown on xylan being increased at least 20-fold by comparison with cells grown on glucose. Ten bacteriophage clones expressing xylanase activity were isolated from a lambda EMBL3 genomic DNA library of P. ruminicola B1(4). These clones were shown to represent four distinct chromosomal regions, based on restriction enzyme analysis and DNA hybridisation. Three groups of clones encoded activity against oat spelt xylan but not carboxymethylcellulose (CMC). In one of these groups, represented by clone 5, activities against pNP-arabinofuranoside and pNP-xyloside were found to be encoded separately from endoxylanase activity. The fourth region encoded activity against CM cellulose and lichen, in addition to xylan, and contains an endoglucanase/xylanase gene isolated previously.
ISSN:0378-1097