• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of aemPermissionsJournals.ASM.orgJournalAEM ArticleJournal InfoAuthorsReviewers
Appl Environ Microbiol. Sep 1995; 61(9): 3288–3292.
PMCID: PMC167609

The cellular location of Prevotella ruminicola beta-1,4-D-endoglucanase and its occurrence in other strains of ruminal bacteria.


Prevotella ruminicola B(1)4, TC1-1, TF1-3, and TS1-5 all produced immunologically cross-reacting 88- and 82-kDa carboxymethyl cellulases (CMCases). P. ruminicola 23, 118B, 20-63, and 20-78 had much lower CMCase activities, and Western blots (immunoblots) showed no cross-reaction with the B(1)4 CMCase antiserum. Fibrobacter succinogenes S85 and Selenomonas ruminantium HD4 and D produced CMCase, but these enzymes were smaller and did not cross-react with the B(1)4 CMCase antiserum. The B(1)4 CMCase antiserum inhibited the B(1)4, TC1-1, TF1-3, and TS1-5 CMCase activities and agglutinated these cells, but it had no effect on the other strains or species. On the basis of these results, the B(1)4 CMCase is a strain-specific enzyme that is located on the outside surface of the cells. P. ruminicola B(1)4 cultures, grown on sucrose, did not have significant CMCase activity, but these cells could bind purified 88- and 82-kDa CMCase but not 40.5-kDa CMCase. Because the 40.5-kDa CMCase is a fully active, truncated form of the CMCase, it appears that the N-terminal domain of the 88-kDa B(1)4 CMCase anchors the CMCase to the cells. Cells grown on cellobiose produced at least 10-fold more CMCase than the sucrose-grown cells, and the cellobiose-grown cells could only bind 15% as much CMCase as sucrose-grown cells. Virtually all of the CMCase activity of exponentially growing cultures was cell associated, but CMCase activity was eventually detected in the culture supernatant. On the basis of the observation that the 88-kDa CMCase was gradually converted to the 82-kDa CMCase when cultures reached the stationary phase without a change in specific activity, it appears that the 82-kDa protein is probably a proteolytic degradation product of the 88-kDa CMCase.

Full Text

The Full Text of this article is available as a PDF (467K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Avgustin G, Flint HJ, Whitehead TR. Distribution of xylanase genes and enzymes among strains of Prevotella (Bacteroides) ruminicola from the rumen. FEMS Microbiol Lett. 1992 Dec 1;78(2-3):137–143. [PubMed]
  • Avgustin G, Wright F, Flint HJ. Genetic diversity and phylogenetic relationships among strains of Prevotella (Bacteroides) ruminicola from the rumen. Int J Syst Bacteriol. 1994 Apr;44(2):246–255. [PubMed]
  • Engelman DM, Steitz TA. The spontaneous insertion of proteins into and across membranes: the helical hairpin hypothesis. Cell. 1981 Feb;23(2):411–422. [PubMed]
  • Gardner RG, Wells JE, Russell JB, Wilson DB. The effect of carbohydrates on the expression of the Prevotella ruminicola 1,4-beta-D-endoglucanase. FEMS Microbiol Lett. 1995 Jan 15;125(2-3):305–310. [PubMed]
  • Henderson R, Unwin PN. Three-dimensional model of purple membrane obtained by electron microscopy. Nature. 1975 Sep 4;257(5521):28–32. [PubMed]
  • Kyte J, Doolittle RF. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Maglione G, Matsushita O, Russell JB, Wilson DB. Properties of a genetically reconstructed Prevotella ruminicola endoglucanase. Appl Environ Microbiol. 1992 Nov;58(11):3593–3597. [PMC free article] [PubMed]
  • Mannarelli BM, Ericsson LD, Lee D, Stack RJ. Taxonomic relationships among strains of the anaerobic bacterium Bacteroides ruminicola determined by DNA and extracellular polysaccharide analysis. Appl Environ Microbiol. 1991 Oct;57(10):2975–2980. [PMC free article] [PubMed]
  • Matsushita O, Russell JB, Wilson DB. Cloning and sequencing of a Bacteroides ruminicola B(1)4 endoglucanase gene. J Bacteriol. 1990 Jul;172(7):3620–3630. [PMC free article] [PubMed]
  • Matsushita O, Russell JB, Wilson DB. A Bacteroides ruminicola 1,4-beta-D-endoglucanase is encoded in two reading frames. J Bacteriol. 1991 Nov;173(21):6919–6926. [PMC free article] [PubMed]
  • Ozols J, Gerard C. Covalent structure of the membranous segment of horse cytochrome b5. Chemical cleavage of the native hemoprotein. J Biol Chem. 1977 Dec 10;252(23):8549–8553. [PubMed]
  • Russell JB. Fermentation of cellodextrins by cellulolytic and noncellulolytic rumen bacteria. Appl Environ Microbiol. 1985 Mar;49(3):572–576. [PMC free article] [PubMed]
  • Vercoe PE, Gregg K. DNA sequence and transcription of an endoglucanase gene from Prevotella (Bacteroides) ruminicola AR20. Mol Gen Genet. 1992 May;233(1-2):284–292. [PubMed]
  • Whitehead TR. Analyses of the gene and amino acid sequence of the Prevotella (Bacteroides) ruminicola 23 xylanase reveals unexpected homology with endoglucanases from other genera of bacteria. Curr Microbiol. 1993 Jul;27(1):27–33. [PubMed]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)


Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...