Assessment of the endo-1,4-beta-glucanase components of Ruminococcus flavefaciens FD-1

Doerner, Kinchel C.; White, Bryan A.

doi:10.1128/aem.56.6.1844-1850.1990

Cited by 51 publications

(28 citation statements)

References 39 publications

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“…Recently Cavedon et al [6] have shown that a free-living, mesophilic Clostridiurn (strain C7), secretes an extracellular 700 000-M r multiprotein cellulolytic complex into the culture supernatant. R. flaeefaciens FD-1 secretes two multiprotein complexes in which all the endoglucanases had xylanase activity [5].…”

Section: Resultsmentioning

confidence: 99%

“…Other bacteria appear to secrete their cellulases into the culture fluid. Two enzyme complexes containing 18 endoglucanases have been found in the culture fluid of the rumen anaerobe, R. flavefaciens FD-1 [5], while Clostridium strain C7 and Cellulomonas fimi produce extracellular cellulase complexes [6,7]. Naturally occurring cellulose is usually associated with hemicellulose.…”

Section: Introductionmentioning

confidence: 99%

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An analysis of the extracellular xylanases and cellulases of Butyrivibrio fibrisolvens H17c

Lin

1991

FEMS Microbiology Letters

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The extracellular xylanase and cellulase components of Butyrivibrio fibrisolvens H17c were investigated. Two major peaks of enzyme activity were eluted by hydroxylapatite chromatography and designated complex A (CA), having cellulase activity, and complex B (CB) having predominantly xylanase activity but with some activity on carboxymethyl cellulose (CMC). CB was further purified on a DE-52 column and subjected to gel filtration. The xylanase and CMCase activities eluted in a single peak with an apparent molecular mass greater than thyroglobulin (Mr 669,000). CMC xymograms of polyacrylamide gels electrophoresed under non-denaturing conditions indicated the presence of five bands with CMCase activity from CA and eight from CB. Xylan xymograms under the same conditions indicated the presence of four bands of activity in CB. Under mild denaturing conditions the xylanase activity in CB was found in 11 bands with molecular mass ranging from 45 to 180 and the CMCase activity in three bands with molecular mass ranging from 45 kDa to 60 kDa. This indicates that CB exists as a multi-subunit protein aggregate of xylanases, some of which also have cellulase activity.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

An analysis of the extracellular xylanases and cellulases of Butyrivibrio fibrisolvens H17c

Lin

1991

FEMS Microbiology Letters

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“…If CMCase negative mutants grew as rapidly on cellulose as wild-type strains and were unable to grow rapidly on L-glu-cans, the claim that non-cellulose-binding CMCases are involved in true cellulose digestion would be dif-¢cult to justify. However, F. succinogenes S85 [32] and cellulolytic ruminococci [2,4,9,10,17] produce a variety of di¡erent CMCases, and this characteristic would confound straightforward mutant selection schemes.…”

Section: Diet Cmcase+ Cmcase3mentioning

confidence: 99%

The role of ruminal carboxymethylcellulases in the degradation of β-glucans from cereal grain

Fields¹

1998

FEMS Microbiology Ecology

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The non-cellulolytic ruminal bacterium, Prevotella bryantii, grew rapidly on water soluble L-glucans, but a mutant deficient in carboxymethylcellulase (CMCase) activity could not. Native activity gels prepared with CMC and L-glucan indicated that the CMCase and L-glucanase activities migrated at similar rates. When a plasmid carrying the P. bryantii CMCase was transferred to Escherichia coli, the clone had CMCase and L-glucanase activities. P. bryantii grew on mixed L-1,3-1,4 glucans, but not on L-1,3 glucan, and similar results were obtained with the cellulolytic ruminal bacteria (Fibrobacter succinogenes S85, Ruminococcus flavefaciens FD1 and Ruminococcus albus B199). Mixed ruminal bacteria from cattle fed hay had twice as much CMCase activity as bacteria from cattle fed 90% cereal grain (P 6 0.05), and the CMCase and L-glucanase activities were highly correlated (r P = 0.93) for the 22 samples tested. The CMCase and L-glucanase activities of mixed ruminal bacteria migrated slowly through polyacrylamide gels, but the migration distances were approximately the same. When L-glucan-utilizing ruminal bacteria were isolated from cattle fed hay or 90% cereal grain, 70 and 38% of the strains, respectively, had CMCase activity. A similar trend was observed with cellobiose-utilizing isolates (70 and 35%, respectively, were CMCase positive). All CMCase positive, cellobiose-utilizing ruminal bacteria could grow on L-glucan. CMCase activity was not strongly correlated with cellulose utilization, and less than 15% of the CMCase positive isolates grew on ball-milled cellulose. Based on these results, the cell-associated CMCases of ruminal bacteria provide a mechanism for utilizing water soluble L-glucans from cereal grains. z

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“…Ruminococcus flavefaciens is a ruminant bacterium highly active on plant cell walls. Two of its strains, R. flavefaciens 17 and R. flavefaciens FD-1, have been extensively studied (Doerner et al, 1990;Doerner et al, 1992;Flint et al, 1993;Vercoe et al, 1995). Both R.…”

Section: Introductionmentioning

confidence: 99%

“…The GH44 EG in this paper, CelB from R. flavefaciens FD-1, has been partially studied (Doerner et al, 1990); it is active on CMC (10 U/mg), oat spelt xylan (3.5 U/mg), and [ 14 C] cellulose (0.0036 U/mg). Temperature and pH optima for CelB have not been published.…”

Section: Introductionmentioning

confidence: 99%

Structures, kinetics, and modeling of glycoside hydrolase family 44 endoglucanases from Clostridium acetobutylicum and Ruminococcus flavefaciens FD-1

Warner

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A gene encoding a glycoside hydrolase family 44 (GH44) protein from Clostridium acetobutylicum ATCC 824 was synthesized and transformed into Escherichia coli. The previously uncharacterized protein was expressed with a C-terminal His-tag and purified by nickel-nitrilotriacetic acid affinity chromatography. Crystallization and x-ray diffraction to 2.2-Å resolution revealed a TIM barrel-like structure with additional Greek key and β-sandwich folds, similar to other GH44 crystal structures. The enzyme hydrolyzes cellotetraose and larger cellooligosaccharides, yielding an unbalanced product distribution including some glucose. It attacks carboxymethylcellulose and xylan at approximately the same rates. Its activity on carboxymethylcellulose is much higher than that of the isolated C. acetobutylicum cellulosome. It also extensively converts lichenan to oligosaccharides of intermediate size and attacks Avicel to a limited extent. The enzyme has an optimal temperature in a 10-min assay of 55°C and an optimal pH of 5.0.

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Assessment of the endo-1,4-beta-glucanase components of Ruminococcus flavefaciens FD-1

Cited by 51 publications

References 39 publications

An analysis of the extracellular xylanases and cellulases of Butyrivibrio fibrisolvens H17c

An analysis of the extracellular xylanases and cellulases of Butyrivibrio fibrisolvens H17c

The role of ruminal carboxymethylcellulases in the degradation of β-glucans from cereal grain

Structures, kinetics, and modeling of glycoside hydrolase family 44 endoglucanases from Clostridium acetobutylicum and Ruminococcus flavefaciens FD-1

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