• Asian-Australasian Journal of Animal Sciences
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• v.12 no.1
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• pp.139-147
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• 1999

Peptides are formed in the rumen as the result of microbial proteinase activity. The predominant type of activity is cysteine ptoteinase, but others, such as serine proteinases, are also present. Many species of protozoa, bacteria and fungi are involved in ptoteolysis; large animal-to-animal variability is found when proteinase activities in different animals are compared. The peptides formed from proteolysis are broken down to amino acids by peptidases. Different peptides are broken down at different rates, depending on their chemical composition and particularly their N-terminal structure. Indeed, chemical addition to the N-terminus of small peptides, such as by acetylation, causes the peptides to become stable to breakdown by the rumen microbial population; the microorganisms do not appear to adapt to hydrolyse acetylated peptides even after several weeks exposure to dietary acetylated peptides, and the amino acids present in acetylated peptides are absorbed from the small intestine. The amino acids present in some acetylated peptides remain available in nutritional trials with rats, but the nutritive value of the whole amino acid mixture is decreased by acetylation. The genus Prevotella is responsible for most of the catabolic peptidase activity in the rumen, via its dipeptidyl peptidase activities, which release dipeptides rather than free amino acids from the N-terminus of oligopeptides. Studies with dipeptidyl peptidase mutants of Prevotella suggest that it may be possible to slow the rate of peptide hydrolysis by the mixed rumen microbial population by inhibiting dipeptidyl peptidase activity of Prevotella or the rate of peptide uptake by this genus. Peptides and amino acids also stimulate the growth of rumen microorganisms, and are necessary for optimal growth rates of many species growing on tapidly fermented substrates; in rich medium, most bacteria use pre-formed amino acids for more than 90% of their amino acid requirements. Cellulolytic species are exceptional in this respect, but they still incorporate about half of their cell N from pre-formed amino acids in rich medium. However, the extent to which bacteria use ammonia vs. peptides and amino acids for protein synthesis also depends on the concentrations of each, such that preformed amino acids and peptides are probably used to a much lesser extent in vivo than many in vitro experiments might suggest.

• Asian-Australasian Journal of Animal Sciences
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• v.28 no.10
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• pp.1433-1441
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• 2015

This study examined changes of rumen fermentation, ruminal bacteria biodiversity and abundance caused by nitrate addition with Ion Torrent sequencing and real-time polymerase chain reaction. Three rumen-fistulated steers were fed diets supplemented with 0%, 1%, and 2% nitrate (dry matter %) in succession. Nitrate supplementation linearly increased total volatile fatty acids and acetate concentration obviously (p = 0.02; p = 0.02; p<0.01), butyrate and isovalerate concentration numerically (p = 0.07). The alpha (p>0.05) and beta biodiversityof ruminal bacteria were not affected by nitrate. Nitrate increased typical efficient cellulolytic bacteria species (Ruminococcus flavefaciens, Ruminococcus ablus, and Fibrobacter succinogenes) (p<0.01; p = 0.06; p = 0.02). Ruminobactr, Sphaerochaeta, CF231, and BF311 genus were increased by 1% nitrate. Campylobacter fetus, Selenomonas ruminantium, and Mannheimia succiniciproducens were core nitrate reducing bacteria in steers and their abundance increased linearly along with nitrate addition level (p<0.01; p = 0.02; p = 0.04). Potential nitrate reducers in the rumen, Campylobacter genus and Cyanobacteria phyla were significantly increased by nitrate (p<0.01; p = 0.01).To the best of our knowledge, this was the first detailed view of changes in ruminal microbiota by nitrate. This finding would provide useful information on nitrate utilization and nitrate reducer exploration in the rumen.

• Applied Biological Chemistry
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• v.37 no.4
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• pp.226-233
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• 1994

Three hundred and one cellulolytic bacterial were isolated from the 148 screening sources such as decomposed wood, soil, compost and leaf mold. Among them, strain KL-6 was found to have the highest of cellulase activity, and identified as species belonged to the genus Cellulomonas. Strain KL-6 was decompose up to 90% of the filter paper (whatman No. 1) substrate within 50 hours, and showed the colony halo formation (11 cm). The activities of CMCase (67 unit/ml), FPase (70 unit/ml) and ${\beta}-glucosidase$ (0.68 unit/ml) were obtained when this strain was cultured for 50 hrs at $30^{\circ}C$. Glucose was not found in detectable amounts at the FP medium. The optimum composition of nutrient medium for the cell growth by strain KL-6 was sucrose 0.5%, yeast extract 0.1%, $(NH_4)_2HPO_4\;0.1%$, $K_2HPO_4\;0.1%$, $MgSO_4{\cdot}7H_2O\;0.01%$, $CaCl_2\; 0.01%$, NaCl 0.6%, $CaCO_3\;0.1%$ and the optimum pH and temperature were 7.0 and $30^{\circ}C$, respectively.

• Applied Biological Chemistry
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• v.28 no.2
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• pp.98-105
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• 1985

In the sheep and cattle's rumen, facultative anaerobic cellulolytic bacteria were isolated by using Hungate's roll tube technique. In the 21 isolated species, one was screened by its strong cellulolytic activity and identified as Cellulomonas fimi C-14 by investigate morphological, cultural, physiological characteristics and electron microgram. Optimum conditions of the cell growth and enzyme production were pH 6.5 an $30^{\circ}C$, Thiamine and biotin support a good growth of C. fimi C-14. In the enzyme activities, Crystalline cellulose hydrolyzing activity, CMCase activity and ${\beta}-glucosidase$ activity were 20.6, 226.6 and 0.56$(unit{\times}10^3/ml)$ at pH 6.0, $40^{\circ}C$. By addition of fungal cellulase, enzyme activity was increased. Simultaneous Saccharification Fermentation is better than two step fermentation in ethanol yield with Saccharomyces cerevisiae DY2.

• Journal of Microbiology and Biotechnology
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• v.17 no.8
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• pp.1291-1299
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• 2007

Two ${\beta}$-1,4-glucanases (DI and DIII fractions) were purified to homogeneity from the culture filtrate of a cellulolytic bacteria, Cellulomonas sp. CS 1-1, which was classified as a novel species belonging to Cellulomonas uda based on chemotaxanomic and phylogenetic analyses. The molecular mass was estimated as 50,000 Da and 52,000 Da for DI and DIII, respectively. Moreover, DIII was identified as a glycoprotein with a pI of 3.8, and DI was identified as a non-glycoprotein with a pI of 5.3. When comparing the ratio of the CMC-saccharifying activity and CMC-liquefying activity, DI exhibited a steep slope, characteristic of an endoglucanase, whereas DIII exhibited a low slope, characteristic of an exoglucanase. The substrate specificity of the purified enzymes revealed that DI efficiently hydrolyzed CMC as well as xylan, whereas DIII exhibited a high activity on microcrystalline celluloses, such as Sigmacells. A comparison of the hydrolysis patterns for pNP-glucosides (DP 2-5) using an HPLC analysis demonstrated that the halosidic bond 3 from the nonreducing end was the preferential cleavage site for DI, whereas bond 2, from which the cellobiose unit is split off, was the preferential cleavage site for DIII. The partial N-terminal amino acid sequences for the purified enzymes were $^1Ala-Gly-Ser-Thr-Leu-Gln-Ala-Ala-Ala-Ser-Glu-Ser-Gly-Arg-Tyr^{15}$-for DI and $^1Ala-Asp-Ser-Asp-Phe-Asn-Leu-Tyr-Val-Ala-Glu-Asn-Ala-Met-Lys^{15}$-for DIII. The apparent sequences exhibited high sequence similarities with other bacterial ${\beta}$-1,4-glucanases as well as ${\beta}$-1,4-xylanases.