• The Korean Journal of Mycology
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• v.21 no.1
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• pp.28-37
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• 1993

An endo-type cellulase, carboxymethyl cellulase(CMCase) IV, was purified from culture filtrate of cellulolytic fungus Penicillium verruculosum. The CMCase IV was acidic glycoprotein having carbohydrate of 13% as glucose and pI value of 4.0. The CMCase IV was 52 KDa of molecular weight in SDS-polyacrylamide gel electrophoresis and have optimum temperature and pH of $50^{\circ}C$ and 5.0 for enzyme activity. The CMCase IV liberated glucose and cellobiose as major products of the enzyme against carboxymethyl cellulose(CMC) and seemed to has transglycosylation activity simultaneously. Cellulase activity staining(zymogram) showed that the cellulase components of P. verruculosum were not aggregated in the medium. P. vrttuculosum mRNA was translated in vitro and translation product by the mRNA coding for CMCase IV was identified by immunoprecipitation.

• Korean Journal of Organic Agriculture
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• v.20 no.3
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• pp.327-343
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• 2012

In order to develop a high cellulolytic direct-fed microorganism (DFM) for ruminant productivity improvement, this study isolated cellulolytic bacteria from the rumen of Holstein dairy cows, and compared their cellulolytic abilities via DM degradability, gas production and cellulolytic enzyme activities. Twenty six bacteria were isolated from colonies grown in Dehority's artificial (DA) medium with 2% agar and cultured in DA medium containing filter paper at $39^{\circ}C$ for 24h. 16s rDNA gene sequencing of four strains from isolated bacteria showed that H8, H20 and H25 strains identified as Ruminococcus flavefaciens, and H23 strain identified as Fibrobacter succinogenes. H20 strain had higher degradability of filter paper compared with others during the incubation. H8 (R. flavefaciens), H20 (R. flavefaciens), H23 (F. succinogenes), H25 (R. flavefaciens) and RF (R. flavefaciens sijpesteijn, ATCC 19208) were cultured in DA medium with filter paper as a single carbon source for 0, 1, 2, 3, 4 and 6 days without shaking at $39^{\circ}C$, respectively. Dry matter degradability rates of H20, H23 and H25 were relatively higher than those of H8 and RF since 2 d incubation. The cumulative gas production of isolated cellulolytic bacteria increased with incubation time. At every incubation time, the gas production was highest in H20 strain. The activities of carboxymethylcellulase (CMCase) and Avicelase in the culture supernatant were significantly higher in H20 strain compared with others at every incubation time (p<0.05). Therefore, although further researches are required, the present results suggest that H20 strain could be a candidate of DFM in animal feed due to high cellulolytic ability.

• Korean Journal of Microbiology
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• v.14 no.2
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• pp.65-74
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• 1976

Atternaria sp. was isolated from soil and crude cellulases were prepared from wheat bran culture of the fungus. The activities of the crude enzyme were studied on five different subvstrates and some phsical properties were also examined, crude enzymes were purified by column chromatography on DEAE Sephadex and Sephadex, Isozymes were separated some of which were active specifically on DEAE cellulose and some were primarily active on cellulose and CM-cellulose. The optimal points of pH and temperature for the crude enzyme were varied depending on the substrates ; On cellulose they were at pH 6.0 and 40.deg.C, on CM-cellulose at pH's 4.0 and 6.0 and 60.deg.C, and on DEAW-cellulose at pH 5.0 and 50.deg.C. Two active fractions, F-1 and F-II on Na-CMC was used as substrate the Km values of crude enzyme, F-I and F-II were calculated to be $4{\times}10^{-5}$ , 1.1 * 10$^{-4}$ , and $1.25{\times}10^{-4}mN$ resepctively. The Ki value of $Cu^{++}$ for crude enzyme was$4{\times}^{-4}mN$ , while that of $Nm^{++}$ while in the same concentration of $Mn^{++}$ it reached to 91%. Some 57% activity of F-1 was inhibited in s mN $Cu^{++}$, whereas it was inhibited as much as 81% in the same concentration above the concentration of 0.3 mM with tis activity reaching up to 137% in 2 mM. On the other hand the F-11 was inhibited by the presence of M $n^{++}$ and some 67% activity was inhibited at 2mM.

• KSBB Journal
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• v.28 no.1
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• pp.42-47
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• 2013

This study was performed to find cellulolytic strain of enzymatic saccharification for bioethanol production. Cellulolytic strains were isolated from 59 different feces of herbivores from Seoul Grand Park located in Gwacheon Gyeonggi-Do. The celluloytic strain was selected by congo red staining and DNS method. Among the isolated strains, H9-1 strain isolated from the feces of rabbit has the highest CMCase activity. H9-1 strain was identified as Bacillus sp. based on 16S rDNA gene sequencing. The optimal conditions for CMCase activity by Bacillus sp. H9-1 were at $40^{\circ}C$ and at initial pH 8.

• Applied Biological Chemistry
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• v.38 no.6
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• pp.490-495
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• 1995

Among the cellulases by Cellulomonas sp. KL-6. CMCase and filter paperase, which were produced as the out enzymes of cell, had been much produced, but very small amounts of ${\beta}-glucosidase $, the enzyme of which is cell bound form, was produced by this organism. The optimal culture times for CMCase and filter paperase productions were 5 days, while that of ${\beta}-glucosidase$ was 4 days. When this strain was cultured under the optimal medium for enzyme production, CMCase, FPase and ${\beta}-glucosidase$ were $82\;units/m{\ell},\;80\;units/m{\ell}\;and\;1.2\;units/m{\ell}$, respectively. Thus these results were showed to increase enzyme productivities as about $60{\sim}70%$ than those produced in basal medium. $CaCO_3$ injected to the medium as the ratio of 0.1% was not only enhanced cellulase activities but also effective as acid neutralizing agent. The production effects of lignase and lactase by this bacterium in filter paper medium was not appeared.

• Korean Journal of Microbiology
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• v.22 no.4
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• pp.207-214
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• 1984

Improved methods for the isolation and purification of conidial protoplast were investigated and conidial proplast from auxotrophic mutants were fused. The reaction time for isolation of protoplasts from the swollen condiospores preincubated in liquid minimal medium supplimented with 2-deoxy-D-glucose was shorten by reaction with mixture of 2% driselase and 2% ${\beta}-glucuronidase$ (1:1). The conidial protoplast could be highly purified by using 5% Ficoll 400 as a centrifugation medium. Nucleus of the conidial protoplast was stained with Giemsa stain and the conidial protoplast had one nucleus. It was also confirmed that the conidial protoplast was true protoplast with no cell wall remnant at the outside of plasma menbrane. Fusion frequencies of the conidial protoplast from auxotrophic mutants ranged from $3.4{\times}10^{-1}\;to\;4.9{\times}10^{-1}$. These values were higher than those of mycelial protoplast by a factor of 5 to 28.

• Korean Journal of Soil Science and Fertilizer
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• v.45 no.4
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• pp.515-523
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• 2012

The study investigated the biochemical methane potential (BMP) assay of cellulose supplementing with mixed methanogens and cellulolytic bacteria to improve anaerobic digestion for methane production. For the BMP assay, 7 different microbial supplementation groups were consisted of the cultures of mixed methanogens (M), Fibrobacter succinogenes (FS), Ruminococcus flavefaciensn (RF), R. albus (RA), RA+FS and M+RA+FS including control. The cultures were added in the batch reactors with the increasing dose levels of 1% (0.5 mL), 3% (1.5 mL) and 5% (2.5 mL). Incubation for the BMP assay was carried out for 40 days at $38^{\circ}C$ and anaerobic digestate obtained from an anaerobic digester with pig slurry as inoculum was used. In results, 5% FS increased total biogas and methane production up to 10.4~22.7% and 17.4~27.5%, respectively, compared to other groups (p<0.05). Total solid (TS) digestion efficiency showed a similar trend to the total biogas and methane productions. Generally the TS digestion efficiency of the FS group was higher than that of other groups showing at the highest value of 64.2% in the 5% FS group. Volatile solid (VS) digestion efficiencies of 68.4 and 71.0% in the 5% FS and the 5% RF were higher than other groups. After incubation, pH values in all treatment groups were over 6.4 indicating that methanogensis was not inhibited during the incubation. In conclusion, the results indicated that the hydrolysis stage for methane production in anaerobic batch reactors was the late-limiting stage compared with the methanogenesis stage, and especially, as the supplementation levels of F. succinogenes supplementation increased, the methane production was increased in the BMP assay compared with other microbial culture addition.

• Journal of The Korean Society of Grassland and Forage Science
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• v.33 no.4
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• pp.304-312
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• 2013

This study was conducted to investigate the effect of total neutral detergent fiber (NDF) and forage-derived NDF (fNDF) on nutrient intake and milk production in Holstein dairy cows. Thirty-six Holstein dairy cows at weeks 4 to 20 of lactation were included in this study. The cows were arranged in a randomized complete block design and assigned to one of six diets: 1) 38% NDF with 18% fNDF (HN-HF); 2) 38% NDF with 15% fNDF (HN-MF); 3) 38% with 12% fNDF (HN-LF); 4) 34% NDF with 18% fNDF (LN-HF); 5) 34% NDF with 15% fNDF (LN-MF); and 6) 34% with 12% fNDF (LN-LF). The cows were fed a total mixed ration twice daily (07:00 and 18:00 h) and given ad libitum access to feed and water throughout the experiment. Data on the intake of nutrients (DM, NEl, CP, NDF, fNDF, NFC), milk yield, composition, fat-corrected milk (FCM), fat and protein corrected milk (FPCM), and energy corrected milk (ECM) were analyzed. Lower dietary concentrations of total NDF or fNDF resulted in an increase in DM, NEl, and NFC intake (p<0.01), milk yield (p<0.05) and a lower milk fat concentration (p<0.01). The LN treatment showed no significant differences in milk fat concentration compared to the HN however, further reduction in fNDF decreased milk fat content (p<0.01). The lowest level of milk fat was observed in the LN-LF diet group. Changes in the dietary concentrations of NDF or fNDF did not affect the concentrations of milk protein or SNF (solid not fat), although the yield of milk protein and SNF increased with higher milk yield (p<0.01). The level of fNDF influenced the level of FCM (p=0.07), as well as FPCM and ECM yield (p<0.05). A highly positive correlation between fNDF intake and milk fat concentration was observed in animals with low NDF compared to those offered high NDF. A strong correlation was also observed between milk yield and low NDF intake compared to high NDF intake.

• The Korean Journal of Mycology
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• v.13 no.4
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• pp.213-219
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• 1985

Among the eight strains, Pleurotus sajor-caju JAFM 1017 was selected as most potent producer of cellulolytic enzymes. The avicelase and CMCase activity reached maximum levels after 10 days, and ${\beta}-glucosidase$ activity reached a maximum level after 19 days. Among the various carbon sources, cellulose powder was most effective for the production of avicelase and ${\beta}-glucosidase$, and Na-CMC (sodium carboxymethyl cellulose) was good for the production of CMCase. The optimum concentration of cellulose powder was 1.0% (w/v), and glucose (1.0%) completely depressed the production of enzymes. Nitrates were effective for the production of enzymes, but nitrites did not support growth. The production of cellulolytic enzymes increased as the concentration of urea increased. The appropriate concentration of urea was 0.054% (w/v).

• Journal of Animal Environmental Science
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• v.16 no.1
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• pp.81-92
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• 2010

This study was conducted to evaluate the effects of fermented diets including liquid by-products on nutrient digestibility and nitrogen balance in growing pigs. Treatments were 1) CON (basal diet), 2) F (fermented diet with basal diet), 3) KF (fermented diet with basal diet including 30% kale pomace), 4) AF (fermented diet with basal diet including 30% angelica keiskei pomace), 5) CF (fermented diet with basal diet including 30% carrot pomace) and 6) OF (fermented diet with basal diet including 30% grape pomace). A total of 24 pigs (41.74kg average initial body weight, Landrace $\times$ Yorkshire $\times$ Duroc), were assigned to 6 treatments, 4 replicates and 1 pig per metabolic cage in a randomized complete block (RCB) design. Pigs were housed in $0.5\times1.3m$ metabolic cage in a 17d digestibility trial. During the entire experimental period, Digestibility of dry matter (p<0.05) of treatment CON, F and CF were higher than other treatments. In crude protein digestibility, treatment F was higher than treatment AF and GF (p<0.05). Treatment GF showed the lowest digestibility of crude fiber among all treatments (p<0.05). In ether extract digestibility, treatment AF and CF showed higher than other treatments (p<0.05) except KF treatment. CF treatment showed the best digestibility of ash among all treatments (p<0.05). Whereas, For Ca and P digestibility, CF and OF treatments were improved than other treatments (p<0.05). Energy digestibility (p<0.05) of CON, F and CF treatments were higher than KF, AF and GF treatments. In total essential amino acid digestibility, F treatment was improved than AF, CF and GF treatments (p<0.05). In total non-essential amino acid digestibility, F treatment was higher than CON, AF and GF treatments (p<0.05). In total amino acid digestibility, F treatment was higher than AF and CF treatments (p<0.05) and GF treatment showed the lowest digestibility (p<0.05). In fecal nitrogen excretion ratio, GF treatment was greatest among all treatments (p<0.05) and F treatment was decreased than other treatments (p<0.05). In urinary nitrogen excretion ratio, CON and GF treatments showed the lowest among all treatments (p<0.05). In nitrogen retention ratio, CON treatment showed the high and KF treatment showed the lost among all treatments (p<0.05). Therefore, this experiment suggested that fermented diet could improve nutrient and amino acid digestibilities of growing pigs.