• Archives of Pharmacal Research
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• v.21 no.1
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• pp.17-23
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• 1998

Flavonoid glycosides were metabolized to phenolic acids via aglycones by human intestinal microflora producing ${\alpha}$-rhamnosidase, exo-${\beta}$-glucosidase, endo- ${\beta}$-glucosidase and/or ${\beta}$-glucuronidase. Rutin, hesperidin, naringin and poncirin were transformed to their aglycones by the bacteria producing ${\alpha}$-rhamnosidase and ${\beta}$-glucosidase or endo- ${\beta}$-glucosidase, and baicatin, puerarin and daidzin were transformed to their aglycones by the bacteria producing ${\beta}$glucuronidase, C-glycosidase and ${\beta}$-glycosidase, respectively. Anti-platelet activity and cytotoxicity of the metabolites of flavonoid glycosides by human intestinal bacteria were more effective than those of the parental compounds. 3,4-Dihydroxyphenylacetic acid and 4-hydroxyl-phenylacetic acid were more effective than rutin and quercetin on anti-platelet aggregation activity. 2,4,6-Trihydroxybenzaidehyde, quercetin and ponciretin were more effective than rutin and ponciretin on the cytotoxicity for tumor cell lines. We insist that these flavonoid glycosides should be natural prodrugs.

• Journal of Applied Biological Chemistry
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• v.57 no.3
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• pp.197-203
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• 2014

Various parameters such as solvent selection, concentration, solid/liquid ratio, soaking time, temperature, stationary, shaking conditions, and repeated extractions were investigated in order to determine the optimum extraction conditions of ${\beta}$-glucosidase from bagasse fermented by mixed culture of Aspergillus niger NRC 7A and Aspergillus oryzae NRRL 447. Among various solvents tested, non ionic detergents gave the best results than the inorganic or organic salt solutions and distilled water. The optimum conditions for extraction of ${\beta}$-glucosidase were 30 min soaking time at $40^{\circ}C$ under shaking condition at 150 rpm, with solid/liquid ratio 1:15 (w/v), which yielded $2882.74{\pm}95.52U/g$ fermented culture (g fc) of enzyme activity. With repeated washes under the above optimum conditions, the results showed that enzyme extracted in the $1^{st}$ and $2^{nd}$ washes represents about 90% of the total activity.

• Journal of Microbiology and Biotechnology
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• v.22 no.12
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• pp.1714-1723
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• 2012

${\beta}$-Glucosidase is necessary for the bioconversion of glycosidic phytochemicals in food. Two Bifidobacterium strains (Bifidobacterium animalis subsp. lactis SH5 and B. animalis subsp. lactis RD68) with relatively high ${\beta}$-glucosidase activities were selected among 46 lactic acid bacteria. A ${\beta}$-glucosidase gene (bbg572) from B. lactis was shotgun cloned, fully sequenced, and analyzed for its transcription start site, structural gene, and deduced transcriptional terminator. The structural gene of bbg572 was 1,383 bp. Based on amino sequence similarities, bbg572 was assigned to family 1 of the glycosyl hydrolases. To overexpress bbg572 in Bifidobacterium, several bifidobacteria expression vectors were constructed by combining several promoters and a terminator sequence from different bifidobacteria. The maximum activity of recombinant Bbg572 was achieved when it was expressed under its own promoter and terminator. Its enzyme activity increased 31-fold compared with those of its parental strains. The optimal pH for Bbg572 was pH 6.0. Bbg572 was stable at $37-40^{\circ}C$. It hydrolyzed isoflavones, quercetins, and disaccharides with various ${\beta}$-glucoside linkages. Bbg572 also converted the ginsenosides Rb1 and Rb2. These results suggest that this new ${\beta}$-glucosidase-positive Bifidobacterium transformant can be utilized for the production of specific aglycone products.

• Journal of the Korean Wood Science and Technology
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• v.35 no.5
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• pp.100-108
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• 2007

$\beta$-Glucosidase from Laetiporus sulphureus among the enzymes related to lignocellulosic biomass degradation to sugars for using alternative bioethanol production was characterized. The highest activity of $\beta$-glucosidase was obtained on cellobiose at shaking culture. For the characterization and purification of $\beta$-glucosidase culture solution was concentrated and then purified by FPLC using ion exchange and size exclusion column. According to the results of SDS-PAGE, native PAGE and microfluidic system of purified enzyme, protein band was observed at about 132 kDa. Optimal pH and temperature of purified $\beta$-glucosi-dase were 5.0 and $60^{\circ}C$, respectively. In the kinetic properties of $\beta$-glucosidase on various substrates such as sophorose, gentiobiose and cellobiose, $K_m$ was 0.81, 1.07 and 1.70 mM, respectively.

• Journal of Microbiology and Biotechnology
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• v.17 no.5
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• pp.745-752
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• 2007

A $\beta$-glucosidase from the algal lytic bacterium Sinorhizobium kostiense AFK-13, grown in complex media containing cellobiose, was purified to homogeneity by successive ammonium sulfate precipitation, and anion-exchange and gel-filtration chromatographies. The enzyme was shown to be a monomeric protein with an apparent molecular mass of 52 kDa and isoelectric point of approximately 5.4. It was optimally active at pH 6.0 and $40^{\circ}C$ and possessed a specific activity of 260.4 U/mg of protein against $4-nitrophenyl-\beta-D-glucopyranoside$(pNPG). A temperature-stability analysis demonstrated that the enzyme was unstable at $50^{\circ}C$ and above. The enzyme did not require divalent cations for activity, and its activity was significantly suppressed by $Hg^{+2}\;and\;Ag^+$, whereas sodium dodecyl sulfate(SDS) and Triton X-100 moderately inhibited the enzyme to under 70% of its initial activity. In an algal lytic activity analysis, the growth of cyanobacteria, such as Anabaena flos-aquae, A. cylindrica, A. macrospora, Oscillatoria sancta, and Microcystis aeruginosa, was strongly inhibited by a treatment of 20 ppm/disc or 30 ppm/disc concentration of the enzyme.

• Journal of Life Science
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• v.21 no.11
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• pp.1579-1585
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• 2011

Natural anti-diabetic semipurified ethyl acetate fraction was isolated from the submerged-liquid culture of Agaricus blaze Murill (AB) in a medium containing soybean flakes. Hot-water extract of AB (HEAB) was prepared by extraction at $121^{\circ}C$ for 60 min, followed by filtering through a filter presser filled with diatomate. The ${\beta}$-glucan-free HEAB, which was a supernatant fraction from HEAB by precipitation in an 80% ethanol solution, was fractionated into hexane, chloroform, ethyl acetate, and butanol fractions. The inhibition of the ${\alpha}$-glucosidase activity by fractions was 59.0, 17.0, 61.6, and 37.9%, respectively, suggesting that ethyl acetate fraction was the most active. A subfraction having a strong ${\alpha}$-glucosidase inhibitory activity (80.4%) was isolated from the ethyl acetate fraction. This subfraction contained isoflavones (genistin and daidzin) and their conjugates with sugars as potent inhibiters of ${\alpha}$-glucosidase activity. These results suggest that the ethyl acetate fraction or HEAB containing isoflavones and their sugars conjugates could be useful sources for controlling blood sugar levels in humans.

• The Korean Journal of Food And Nutrition
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• v.25 no.3
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• pp.620-628
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• 2012

We examined the effects of biological activity Phellinus linteus mycelium culture with cassiae semen extract. Firstly, the optimal temperature, initial pH and culture period for mycelial growth in a liquid culture of P. linteus were determined, and they were $30^{\circ}C$, pH 5.0 and 8 days respectively. The five herbal materials were examined against several health functional efficacies, and, as a result, Cassiae semen was chosen, with its superior inhibitory effects in ${\beta}$-glucuronidase inhibitory activity, electron donating activity, ACE inhibitory, and ${\alpha}$-glucosidase inhibitory activities(95.3%, 80.9%, 96.1 and 24.2%, respectively). P. linteus fruit body was investigated on ${\beta}$-glucuronidase inhibitory activity, electron donating activity, ACE inhibitory, and ${\alpha}$-glucosidase inhibitory activities, and they were 54.7%, 81.9%, 30.0% and 20.1%, respectively. Accordingly, C. semen was used in the following experiment, to give an additive functional effect on the P. linteus. As the amount of C. semen in the cultural media increased, mycelial weight and ${\beta}$-glucan contents also increased, but final pH was not influenced. In addition, the ${\beta}$-glucuronidase inhibitory activity, electron donating activity, and ${\alpha}$-glucosidase inhibitory activity increased. P. linteus mycelium culture showed higher activities in the other three tests above, except for electron donating activity, when C. semen was added to the medium before cultivation.

• The Korean Journal of Mycology
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• v.12 no.4
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• pp.133-140
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• 1984

Some properties of cellulolytic enzymes produced by Pleurotus sajor-caju JAFM 1017 during its growth in synthetic medium were investigated. The optimum pH of avicelase, CMCase, and ${\beta}-glucosidase$ was pH 5.5, pH 4.5 and pH 6.0, respectively. Avicelase and CMCase were stable within pH 5.0 to 6.0 and 4.0 to 6.0, respectively, and ,${\beta}-glucosidase$ was within pH 5.5 to 6.5. The optimum temperature of avicelase, CMCase and ${\beta}-glucosidase$ was the same of $40^{\circ}C$. The enzymes were stable below the optimum temperature, but the enzymes were unstable over the temperature of $50^{\circ}C$, and avicelase was losing about 91.7% of activity at $70^{\circ}C$ for 10 min. The enzyme activity of avicelase and CMCase was increased in proportion to the substrate concentration within 1% and 0.7%, respectively, and ${\beta}-glucosidase$ was within 0.1%. The Michaelis constants (Km) of avicelase and CMCase were 30.77mg avicel/ml and 14.64m Na-CMC/ml, respectively and ${\beta}-glucosidase$ was 5. 13mg salicin/ml. The reducing sugar production of avicelase was proportionaly increased until 120 min. and CMCase and ${\beta}-glucosidase$ were until 60min. The activity of three cellulolytic enzymes were increased by $Ca^{2+}$ at the concentration of $10^{-2}M$, but were inhibited by $Hg^{2+}$, $Ag^+$.

• Journal of Microbiology and Biotechnology
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• v.22 no.12
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• pp.1705-1713
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• 2012

A gene encoding ${\beta}$-glucosidase was cloned from Weissella cibaria 37, an isolate from human feces. Sequence analysis showed that the gene could encode a protein of 415 amino acids in length, and the translated amino acid sequence showed homology (34-31%) with glycosyl hydrolase family 1 ${\beta}$-glucosidases. The gene was overexpressed in E. coli BL21(DE3) using pET26b(+) and a 50 kDa protein was overproduced, which matched well with the calculated size of the enzyme, 49,950.87 Da. Recombinant ${\beta}$-glucosidase was purified by using a his-tag affinity column. The purified ${\beta}$-glucosidase had an optimum pH and a temperature of 5.5 and $45^{\circ}C$, respectively. Among the metal ions (5mM concentration), $Ca^{2+}$ slightly increased the activity (108.2%) whereas $Cu^{2+}$ (46.1%) and $Zn^{2+}$ (56.7%) reduced the activity. Among the enzyme inhibitors (1 mM concentration), SDS was the strongest inhibitor (16.9%), followed by pepstatin A (45.2%). The $K_m$ and $V_{max}$ values of purified enzyme were 4.04 mM and 0.92 ${\mu}mol/min$, respectively, when assayed using pNPG (p-nitrophenyl-${\beta}$-D-glucopyranoside) as the substrate. The enzyme liberated reducing sugars from carboxymethyl cellulose (CMC).

• The Korean Journal of Mycology
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• v.42 no.1
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• pp.86-90
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• 2014

Button mushroom (Agaricus bisporus) strains from diverse origins were compared in this study to obtain basic information on their growth and biochemical properties that are helpful for breeding. Among 31 dikaryotic strains tested, most strains showed better mycelial growth on oatmeal agar than on MEA and PDA. Mycelia of the mushroom strains revealed ${\beta}$-glucosidase activity the most clearly among the seven extracellular enzymes tested. All the strains showed protease activity, but ${\beta}$-glucosidase activity was found in 27 strains and xylanase activity was found in 30 strains. The activity of avicelase, CM-cellulase, amylase, and pectinase was detected in less than 20 strains. These results implied that enzymatic characteristics could be used as a criterion of strain selection for breeding study.