• Journal of Microbiology and Biotechnology
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• v.21 no.5
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• pp.503-508
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• 2011

A ${\beta}$-glucosidase from Phoma sp. KCTC11825BP isolated from rotten mandarin peel was purified 8.5-fold with a specific activity of 84.5 U/mg protein. The purified enzyme had a molecular mass of 440 kDa with a subunit of 110 kDa. The partial amino acid sequence of the purified ${\beta}$-glucosidase evidenced high homology with the fungal ${\beta}$- glucosidases belonging to glycosyl hydrolase family 3. Its optimal activity was detected at pH 4.5 and $60^{\circ}C$, and the enzyme had a half-life of 53 h at $60^{\circ}C$. The $K_m$ values for p-nitrophenyl-${\beta}$-D-glucopyranoside and cellobiose were 0.3 mM and 3.2 mM, respectively. The enzyme was competitively inhibited by both glucose ($K_i$=1.7 mM) and glucono-${\delta}$-lactone ($K_i$=0.1 mM) when pNPG was used as the substrate. Its activity was inhibited by 41% by 10 mM $Cu^{2+}$ and stimulated by 20% by 10 mM $Mg^{2+}$.

• The Korean Journal of Mycology
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• v.17 no.2
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• pp.57-65
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• 1989

The effect of various cellulosic and hemicellulosic substrates on the induction of cellulase and xylanase complexes in Aapergillus nidulans was investigated. The most efficient substrates for the induction of cellulase and xylanase complexes were carboxymethylcellulose for endoglucanase, cellobiose for ${\beta}-glucosidase$, and xylan for endoxylanase and ${\beta}-xylosidase$, respectively. However, the mixtures of these substrates, especially CMC-xylan and CMC-xylan-laminarin mixture, were much more effective not only for the enhancement of the biosynthesis of all the cellulase and xylanase complexes but also for the balanced production of these enzyme components than individual substrate. The polyacrylamide gel electrophoresis followed by activity staining showed the variation in the patterns and relative intensity of ${\beta}-glucosidase$, endoglucanase and endoxylanase components in individual enzyme preparations from A. nidulans cultures grown on different substrates. These results suggest that the biosynthesis is of cellulase and xylanase systems in A. nidulans is regulated in coordination at the level of induction.

• KSBB Journal
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• v.16 no.2
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• pp.174-178
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• 2001

Soybeans contain the phytoestrogens genistein and daidzein, their glucosides genistin and daidzin and coumesterol. These isoflavonoid compounds are capable of producing an estrogenic response in a number of diverse species. This study determined optimum conditions for the extraction of the main isoflavones(daidzin, genistin, daidzein, genistein) in defatted soybean meal using high-performance liquid chromatography. The best optimum extraction was achieved at 75% ethanol, $80^{circ}C$, pH4 and a three hour contact time. In addition, isoflavones with high purity were separated by adding up to 4%(w/v) of calcium chloride dihydrate. Most soybean extracts were composed of $beta$-glucosidic conjugate(daidzin, genistin) which is difficult to adsorb in body. Therefore, $beta$-glucosidase was used to convert as conjugate to aglycone form (daidzein, genistein) which is easy to adsorb. The optimal conditions of enzyme reaction involved to be 8.4 units of enzyme concentration, pH5.0, $40^{circ}C$ and 40 minutes.

• Korean Journal of Ecology and Environment
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• v.35 no.4 s.100
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• pp.326-330
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• 2002

A wetland soil was sterilised by two methods and changes in microbial enzyme activities were assessed. The short-term effects were determined by toluene addition, while the longer-term effects of elimination was monitored by ${\gamma}$-radiation. The changes in ${\beta}$- glucosidase, ${\beta}$-xylosidase, cellobiohydrolase, phosphatase, arylsulphatase, and N-acetylglucosaminidase activities were determined by using methylumbelliferyl model substrates and comparing with the activities of control samples. Toluene addition induced different responses of enzymes. For example, phosphatase activity increased by the treatment while ${\beta}$-glucosidase and arylsulphatase activities decreased. In contrast, ${\gamma}$-radiation decreased all enzyme activities compared to control by 40-80%. The overall results of the toluene and ${\gamma}$-radiation experiments indicate that the large amounts of enzymes are stabilised outside of living cells, at least in the short term, but that the persistence of enzymes is maintained by de-novo synthesis of microbes.

• KSBB Journal
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• v.27 no.6
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• pp.341-346
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• 2012

Indican (indoxyl-${\beta}$-D-glucoside) is a colorless natural compound and can be used as a precursor for the production of indigo. This production step only require an enzyme, ${\beta}$-glucosidase, that readily screened from microbial resource by using selective media supplemented with indican as a sole carbon source. Agrobacterium tumefaciens was well grown in this media and thus presumed to produce a related enzyme. The corresponding gene, encoding a protein with a calculated molecular mass of 51 kDa, was cloned and overexpressed as MBP fusion proteins. The purified enzyme was determined to be a dimer and showed the maximum activity for indican at pH 7.0 and $40^{\circ}C$. The kinetic parameters for indican, Km and Vmax, were determined to be 1.4 mM and 373.8 ${\mu}M/min/mg$, respectively. The conversion yield of indican into indigo using this enzyme was about 1.7-1.8 folds higher than that of previously isolated enzyme from Sinorhizobium meliloti. Additionally, this enzyme was able to hydrolyze various ${\beta}$-1,4 glycoside substrates.

• Journal of Life Science
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• v.20 no.11
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• pp.1589-1594
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• 2010

Glutaraldehyde was used to cross-link chitosan beads to immobilize the crude enzyme $\beta$-glucosidase from Exiguobacterium sp. DAU5. The conditions for preparing cross-linking chitosan beads and immobilization such as concentration of glutaradehyde, cross-linking time, immobilization pH and time were optimized. The chitosan beads were cross-linked with 1.5% glutaraldehyde for 1.5 hr. The immobilized $\beta$-glucosidase had an overall yield of 20% and specific activity of 5.22 U/g. The optimized pH and temperature were 9.0 and $55^{\circ}C$, respectively. More than 80% of its activity at pH 7.0-10.0, 80% at $40^{\circ}C$ for 2 hr and 48% at $50^{\circ}C$ for 1 hr, were retained. However, the immobilization product showed higher pH and thermal stabilities than free enzymes. It also showed high hydrolyzing activity on soybean isoflavone glycoside linkage. These results suggest the broad application prospects of immobilization enzymes.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.6
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• pp.582-586
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• 2005

In this Study, we have attempted to determine the effects of dietary fructose polymers (fructan), high molecular-weight ${\beta}-(2,6)-linked$ levan, and low-molecular-weight ${\beta}-(2,1)-linked$ inulin, on two intestinal enzymes $({\beta}-glucuronidase\;and\;{\beta}-glucosidase)$. As a preliminary experiment, when intestinal microflora were cultured in anaerobic media harboring levan or its oligosaccharides, bacterial cell growth was observed in the levanoligosaccharide-supplemented media, but not in the levan-supplemented media, indicating that levan's size is important for the utilization by intestinal bacteria of levan as an energy Source. In our animal study, the intake of a levan-rich diet was determined to significantly attenuate the activity of the harmful enzyme $({\beta}-glucuronidase$, but d id not affect the activity of ${\beta}-glucosidase$.

• Applied Biological Chemistry
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• v.12
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• pp.99-105
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• 1969

The yield of cellulase derived from Trichoderma $(O_2-1)$ was remarkably varied with various concentration of ethanol and acetone in purification of the enzyme. In the purification with ethanol of ${\beta}-glucosidase$, the best result was obtained in the concentration of 60% and, of CMCase and of filter paper disintegrating enzyme 80%. And in the purification with acetone of ${\beta}-glucosidase$, filter paper disintegrating enzyme, and CMCase, in the concentration of 60%, 80%, and 90% respectively, was shown the best yield. The activities of crude Cellulase preparation could be seperated into few of fractions by column chromatography with Silica gel, Cellulose powder, and gauze. Most of CMCase, avicelase, and ${\beta}-glucosidase$ were eluted, but most of filter paper disintegrating enzyme and the rest of enzymes mentioned the above were absorbed, and were eluted with water. Therefore, it was considered that CMCase is different from filter paper disintegrating enzyme in properties. The relative activity of CMCase was different from that of avicelase in the peak of elusion part. And it was considered that filter paper disintegrating enzyme and cellulose powder saccharifying enzyme was seperated respectively as absorption part and non absorption part. The auther came to the conclusion that at least there were more than three sorts of cellulase in Trichoderma $(O_2-1)$ cellulase preparation.

• Journal of Life Science
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• v.27 no.2
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• pp.172-179
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• 2017

The fermentation of Panax ginseng can yield many compounds from ginsenosides that have a wide variety of biological functions. Lactic acid bacteria (LAB) strains are capable of converting ginsenosides. The purposes of this study were to: (1) characterize Enterococcus faecium SA5, an isolated LAB from Mongolian mare milk, (2) identify the existence of extracellular ${\beta}$-glucosidase activity in the milk, and (3) ascertain if the ${\beta}$-glucosidase has the capacity of converting ginsenoside in Korean ginseng. The results revealed that E. faecium SA5 was acid-resistant, bile salt-resistant, and has antibiotic activities against 4 pathogenic microorganisms (Salmonella typhimurium KCTC 3216, Listeria monocytogenes KCTC 3710, Bacillus cereus KCTC 1012, Staphylococcus aureus KCTC 1621). In addition, E. faecium SA5 had tolerance against some antibiotics such as colistin, gentamycin and neomycin. It was also found that E. faecium SA5 possessed bile salt hydrolase activity, which could lower blood cholesterol level. When incubated in 10% (w/v) skim milk as a yogurt starter, E. faecium SA5 caused to decrease pH of the medium as well as increase in viable cell counts. Using TLC and HPLC analysis on the samples incubated in MRS broth, our study confirmed that E. faecium SA5 can produce ${\beta}$-glucosidase, which was capable of converting ginsenoside $Rb_1$ into new ginsenosides $Rg_3-s$ and $Rg_3-r$. It was concluded that E. faecium SA5 possessed a potential of probiotic activity, which could be applied to yogurt manufacture as well as ginsenoside conversion in ginseng.

• Food Engineering Progress
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• v.14 no.4
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• pp.292-298
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• 2010

Wheat germ contains the glycosylated forms of 2-methoxy-p-benzoquinone (2-MBQ) and 2,6-dimethoxy-p-benzoquinone (2,6-DMBQ), both of which have antimicrobial and immunostimulatory effects. Conversion of glycosylated 2-MBQ and 2,6-DMBQ to their more functional unglycosylated forms requires enzymatic action of $\beta$-glucosidase. We investigated the applications of lactic acid bacteria and yeast that produce $\beta$-glucosidase as starters for production of unglycosylated 2-MBQ and 2,6-DMBQ from wheat germ. Lactobacillus zeae and Pichia pijperi were selected through $\beta$-glucosidase enzyme assays for 37 yeast strains and five strains of lactic acid bacteria. Lb. zeae was more efficient than P. pijperi at producing 2-MBQ and 2,6-DMBQ from wheat germ. After 48 hr of fermentation with a mixed culture of Lb. zeae and P. pijperi, the concentration of 2-MBQ was 0.46${\pm}$0.07 mg/g, indicating an approximately 1.6-fold higher concentration than that obtained by pure culture of Lb. zeae. However, the concentration of 2,6-DMBQ was not significantly enhanced by fermentation with a mixed culture of Lb. zeae and P. pijperi.