• Mycobiology
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• v.37 no.2
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• pp.121-127
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• 2009

Purification and characterization of intracellular cellulase produced by A. oryzae ITCC-4857.01 are reported. The enzyme was purified by ion-exchange chromatography using DEAE-cellulose followed by Gel filtration. The purification achieved was 41 fold from the crude extract with yield of 27%. The purified enzyme showed single band on poly acrylamide gel. The molecular weight as determined by SDS-PAGE and gel filtration was 38 KDa and 38.6 KDa respectively and contained only one subunit. The enzyme is glycoprotien as nature and contained 0.67% neutral sugar. The apparent Km value of the enzyme against cellulose was 0.83%. The enzyme showed the highest relative ativities on CMC followed by avicel, salicin and filter paper. The optimum pH of activity was 5.5 and very slight activity was observed at or above pH 7.5 as well as bellow pH 3.5. The optimum tempreture of the activity was $45^{\circ}C$ and the highest activity was exhibited in 35 to $45^{\circ}C$. The enzyme lost their activities almost completely (95${\sim}$100%) at $80^{\circ}C$ or above and as well as bellow $25^{\circ}C$.

• Journal of Plant Biology
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• v.36 no.4
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• pp.327-335
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• 1993

The active sites, intracellular transport, function of cellulase in association with the disintegration of the storage materials of the endosperm cells during seed maturation and after-ripening of Panax ginseng C.A. Meyer seeds were studied by electron microscopy. Cytochemical activities of the cellulase occurred in protein bodies and vesicles of endosperm cells in seed with red seed coat. In after-ripening seed, the activities were strongly found in the cell wall of endosperm near the umbiliform layer and on neighbouring vesicles, so it is assumed that these cells begin to be decomposed. Cellulase activities were initiated before the decomposition of storage materials. But, no activity was observed in the umbiliform layer, so it is suggested that cellulase lose its activity after the completion of lysis process.

• The Korean Journal of Mycology
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• v.18 no.4
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• pp.209-217
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• 1990

The influence of cellulosic and hemicellulosic substrates on the production of cellulase and xylanase complexes in Aspergillus niger was investigated. The culture conditions with different substrates exhibited profound effects on the level of endoglucanase (CMCase), ${\beta}-glucosidase$, endoxylanase and ${\beta}-xylosidase$, and on their isozyme patterns. However, intracellular and extracellular isozyme patterns of cellulase and xylanase complexes were qualitatively identical and appeared to be simultaneous in the early growth phase. Prolonged incubation led to the increase in the concentrations of isozymes with a little changes in the relative proportions of those isozymes. These results suggest that the biosynthesis of cellulase and xylanase complexes in A. niger is coordinately regulated at the level of induction. Moreover, multiple forms of extracellular cellulase and xylanase complexes seem to be the outcome of specific gene expression and should not be considered solely as the consequence of post-secretional modification of synthesized enzymes.

• Journal of Ginseng Research
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• v.25 no.1
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• pp.53-58
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• 2001

To clarify a significant difference between red-colored phenomena (RCP) and microbes isolated from rhizosphere soil of healthy ginseng (HES) and red-colored ginseng (RCS), we have examined growth and cellulase activities of the microbes according to pH variation and iron status. The soil microbes could not grow at pH 3.0 on the YEB medium. The growth of bacterium isolated from RCG at pH from 5.0 to 9.0 showed small differences and the growth of bacterium HES was lower than that of others. The growth of bacteria from RCS and surface soil (SUS) at pH 5.0 were also lower than that of pH 7.0 and pH 9.0. However, the bacteria isolated from red-colored ginseng (RCG) and RCS are able to grow on the medium contained 2 mM Fe$\^$3+/ at pH 3.0. Furthermore, the growth of bacterium from RCG increased about two times in the medium contained iron at pH 7.0 compared with minus iron. The cellulase activity of isolated bacteria increased two times in the medium contained 2 mM Fe$\^$3+/ compared with minus iron. The activity of extra-cellular cellulase was higher by one hundred times than that of intracellular level. The cellulase activity of the bacterium from RCS at pH 5.0 was higher by two times than that of pH 7.0. Especially, intracellular activity of the bacterium from RCS on the medium contained 2mM Fe$\^$3+/ increased about six to seven times compared with control (minus iron). Also, extra-cellular activity increased about eleven to twelve times compared with control. These results indicate that the soil microbes seem to be related iron redoxidation by proton extrusion and with cell wall digestion by secreted cellulase.

• Microbiology and Biotechnology Letters
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• v.14 no.5
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• pp.377-383
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• 1986

About 300 cellulolytic actinomycetes isolated from soils were tested for their cellulase activities estimated by means of filter paper swelling and carboxymethyl cellulose saccharifying activity. Then, 16 isolates which had shown relatively high levels of CMCase activity were selected and examined for their abilities of $\beta$-glucosidase production. Among them strain No. 109 was found to have highest level of intracellular $\beta$-glucosidase, and selected for the further studies. In this paper, the cultural, morphological and physiological properties, and cell wall composition of strain No. 109 were described in relation to the taxonomic status of this actinomycete. Based on the results obtained in these experiments strain No. 109 was identified to be a similar species to Streptomyces tanashiensis.

• Korean Journal of Microbiology
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• v.23 no.2
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• pp.77-83
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• 1985

The aim of this investigation was to resolve the contradiction between the results of Sternberg and Mandels (1980, 1982)and those of Nisizawa et al., (1971) in cellulase induction by sophorose, and furthermore to study the conditional effects in sophorose-induced cellulase induction in Trichoderma reesei QM 9414. Sophorose could induce the synthesis of CMCase and ${\beta}-glucosidase$ simultaneously. Optimal induction medium by sophorose had the potassium citrate buffer solution of pH 3.0-4.0 for CMCase, but one of pH 5.0-6.0 for ${\beta}-glucosidase$. At this time, two different types of ${\beta}-glucosidase$ could be induced by sophorose: one was extracellular and had maximum at pH 5.0, the other was intracellular and had maximum activity at pH6.5. Induction study showed that $methyl-{\beta}-glucoside$ was not a true inducer of ${\beta}-glucosidase$ and that large ${\beta}-glucosidase$ induction could be obtained only by the addition of sophorose into the induction medium. Glucose repressed the induction of cellulase by sophorose. The repression of glucose could not be overcome by the addition of cyclic AMP into the induction medium.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 1986.12a
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• pp.529.1-529
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• 1986

For isolation of the CMCase gene of the alkalophilic Bacillus sp. strain N-4 to analyze their genetic information for the multicomponents of the cellulase, Bscherichia coli K12 and plasmid DNA pBR322 was used as host-vector system. After the digestion of purified chromosomal DNA and plasmid DNA pBR322 with HindIII, these were ligated. The ligated DND were transformed into Escherichia coli, and recombinant plasmid 107 carried the gene coding for CMCase was constructed. The CMCase produced by Escherichia coli cells containing plasmid DNA pYBC107 was found in the cells as intracellular enzyme and nearly 60% of the total CMCase activity was localized in cellular fraction. Also, the optimum pH for the reaction of CMCase produced by Escherichia coli was appeared at pH .8.0 and the enzyme was stable between pH 7.0 and pH 8.0.