• Korean Journal of Agricultural Science
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• v.24 no.2
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• pp.275-282
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• 1997

The prcx.iuction of extracellular 1,4-${\beta}$-glucanase by Cellulomonas sp. CS1-1 on microcrystalline cellulose, sigmacell was maximal after 5-day cultivation as 280 units/mL, which was three times higher than the level produced on carboxymethyl cellulose. A carboxymethyl cellulase containing the carbohydrate of 8.2% was purified from the culture filtrate by successive procedures of column chromatographies. Purification factor was calculated as 22-folds with the specific carboxymethyl cellulase activity of 31.9 units/mg. The molecular weight and isoelectric point of the purified enzyme were 54,000 and pI 5.4, respectively. The optimal pH and temperature were 6.0 and $45^{\circ}C$, and the enzyme was stable between pH 6.5 and 7.5 and below $50^{\circ}C$. The estimated Km and Vmax were 10 mg/mL and $6.25{\mu}mol/min$ for carboxymethyl cellulose and 30.3 mg/mL and $2.85{\mu}mol/min$ for sigmacell, respectively. The enzyme was partially inhibited by $Ag^+$, $Zn^{+{+}}$, $Fe^{+{+}}$ and EDTA, while completely inhibited by $Cd^{+{+}}$ and $Hg^{+{+}}$ at 1 mM concentration.

• Journal of Animal Science and Technology
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• v.48 no.3
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• pp.415-424
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• 2006

The purpose of this study was to investigate the fungal growth and enzyme production under different carbohydrate substrate conditions. The anaerobic fungus Neocallimastix sp. NLRI-3 isolated from the rumen of Korean native goat was incubated with different carbohydrate media containing 0.2% of glucose, starch, rice straw, filter paper, carboxymethyl cellulose(CMC), Sigmacell cellulose, xylan or xylose, respectively. The culture head gas production was the highest in the culture of filter paper medium, and the lowest in the culture of CMC medium at 96h incubation (P<0.05). The fungal zoospore production reached peak at 72h incubation, and its number was the highest in rice straw medium among the treatments (P<0.05). At 96h incubation, carboxymethyl cellulase(CMCase) activity was the highest in the culture of filter paper medium and the lowest in the culture of starch medium (P<0.05). While xylanase activity was the highest in the culture of rice straw medium and the lowest in the culture of xylose medium(P<0.05) at 72h incubation. There were no differences in culture supernatant protein expression among the treatments. However, the patterns of enzyme expression were different among the treatments with zymogram analysis. Six CMCases and 4 xylanase were detected from the results of zymogram analysis. Therefore the present study indicating that the fungal enzyme expression could be stimulated with insoluble substrates in the culture medium.

• Journal of Chitin and Chitosan
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• v.23 no.4
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• pp.228-233
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• 2018

We isolated microorganisms from soil, which is sampled at forest, Kyeonbuk, Korea, as cellulolytic microorganisms. The isolated strains were identified by analysis of 16S rRNA gene from the starins. The result, four kinds of Bacillus subtilis, one kind of Bacillus amyloliquefaciens, and one kind of Bacillus cereus were identified. Among these strains, Bacillus subtilis was selected due to its high cellulase activity and this strain was named as Bacillus subtilis CNS. The optimum pH and temperature of the cellulase from Bacillus subtilis CNS was pH 5.0 and $40^{\circ}C$, respectively. In the investigation of pH and temperature stability, the cellulase from Bacillus subtilis NSC stabled pH 4.0~6.0 range and until $40^{\circ}C$ for 30 min perfectly. In the enzyme activity for various cellulosic substrate, cellulase from Bacillus subtilis CNS showed the highest activity for CM-cellulose. And, the enzyme activities for alkali swollen cellulose, Alpha-cellulose, Sigmacell-cellulose, and Avicel were approximately 31%, 8%, 8% and 4% of activity for CM-cellulose, respectively. In the degradation of CM-cellulose, the 0.26 U/ml and 0.52 U/ml of cellulase showed 0.43 and 0.76 U/ml activity for CM-cellulose after the reaction of 120 min, respectively.

• Korean Journal of Agricultural Science
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• v.10 no.2
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• pp.365-370
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• 1983

Cellooligosaccharides were prepared from microcrystalline cellulose by acetolysis, deacetylation, and chromatographic fractionation, to determine the optimum time of treatment with acetolysis mixture. The fractionation was accomplished by ethanol-water gradient elution on chromatographic column composed of stearic acid-treated mixture of charcoal and Celite. The amount of initial acetolysis product was increased with the reaction period, however, the acetylated mixture of longer reaction period appeared to contain the greater bulk of methanol-insoluble material. Better yields of most of oligosaccharides were obtained with 3-hr acetolysis. When the reaction time was extended to 4-hr, yields of shorter chain oligosaccharides increased at the expense of the The yields obtained with 3-hr acetclys is from 68.2g of Sigmacell Type 19 were : cellotriose, 1.36g(2.0%) ; -tetraose, 1.64g(2.4%) ; -pentaose, 1.16g(1.7%) ; -hexaose, 0.82g(1.2%) ; and -heptaose, 0.21g(0.3%).

• Applied Biological Chemistry
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• v.51 no.4
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• pp.299-304
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• 2008

A ${\beta}$-1,4-endoglucanase gene from Bacillus subtilis H12 was cloned into Escherichia coli JM109 (pBC8) and sequenced. The endoglucanase gene with an insert DNA of 2.5 kb possessed an open reading frame of 1,500 bp encoding a mature protein of 499 amino acids with a calculated molecular mass of 55 kDa. The deduced amino acid sequence showed similarity to those of the known neutral cellulase genes of B. subtilis PAP115 (99.2%) and BSE616 (97.8%), as well as the alkaline gene of Bacillus sp. N4 (55.1%). The endoglucanase activity expressed by E. coli (pBC8) was localized in the periplasmic fraction (80%) and the cytoplasmic fraction (20%). An endoglucanase was purified from the periplasmic fraction by performing gel filtration and anion exchange chromatography. The molecular weight of the purified enzyme was estimated to be 31 kDa by SDS-PAGE, and the maximum activity occurred at pH 7 and $40^{\circ}C$. The enzyme easily hydrolyzed soluble substrates such as carboxymethyl cellulose and barely ${\beta}$-glucan, whereas the sigmacell and xylan, the known insoluble substrates, were not entirely hydrolyzed.