Isolation of genes encoding β-D-xylanase, β-D-xylosidase and α-L-arabinofuranosidase activities from the rumen bacterium Prevotella ruminicola B(1)4

A Gasparic, R Marinsek-Logar, J Martin, R J Wallace, F V Nekrep, H J Flint, J C Martin

Research output: Contribution to journalArticle

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Abstract

Prevotella ruminicola B14 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 A EMBL3 genomic DNA library of P. ruminicola B14. 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 lichenan, in addition to xylan, and contains an endoglucanase/xylanase gene isolated previously.
Original languageEnglish
Pages (from-to)135-141
Number of pages7
JournalFEMS Microbiology Letters
Volume125
Issue number2-3
DOIs
Publication statusPublished - 15 Jan 1995

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Prevotella ruminicola
Xylosidases
Endo-1,4-beta Xylanases
Rumen
Xylans
Clone Cells
Bacteria
Genes
Restriction Mapping
Carboxymethylcellulose Sodium
Genomic Library
Cellulase
Gene Library
Cellulose
Bacteriophages
Polysaccharides
Glucose
DNA

Keywords

  • animals
  • blotting, southern
  • DNA, bacterial
  • endo-1,4-beta xylanases
  • gene library
  • genes, fungal
  • glycoside hydrolases
  • kinetics
  • prevotella
  • recombinant proteins
  • restriction mapping
  • rumen
  • substrate specificity
  • xylosidases

Cite this

Isolation of genes encoding β-D-xylanase, β-D-xylosidase and α-L-arabinofuranosidase activities from the rumen bacterium Prevotella ruminicola B(1)4. / Gasparic, A; Marinsek-Logar, R; Martin, J; Wallace, R J; Nekrep, F V; Flint, H J; Martin, J C.

In: FEMS Microbiology Letters, Vol. 125, No. 2-3, 15.01.1995, p. 135-141.

Research output: Contribution to journalArticle

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abstract = "Prevotella ruminicola B14 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 A EMBL3 genomic DNA library of P. ruminicola B14. 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 lichenan, in addition to xylan, and contains an endoglucanase/xylanase gene isolated previously.",
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AU - Nekrep, F V

AU - Flint, H J

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AB - Prevotella ruminicola B14 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 A EMBL3 genomic DNA library of P. ruminicola B14. 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 lichenan, in addition to xylan, and contains an endoglucanase/xylanase gene isolated previously.

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KW - restriction mapping

KW - rumen

KW - substrate specificity

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