• Journal of Plant Biology
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• v.39 no.2
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• pp.93-98
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• 1996

In order to enhance the system of potato transformation and further regeneration, potato was transformed using the Agrobacterium tumefaciens harboring $\beta$-glucuronidase (GUS) gene. We found that a series fo modified medium ttained 100% shoot regeneration within 5 weeks after the preincubated explants on stage I medium were infected with Agrobacterium. Callus appeared at the cut edges of stem segments on stage II medium, mainly at the basal parts. Some explants started to form shoots after two to three weeks on stage III medium containing kanamycin (50 mg/L). When transferred to MS medium containing 200 mg/L kanamycin, 81% of the transformed shoots formed roots at the cut edge of the plantlets. In contrast, untrasformed shoots never rooted and became yellowish after few weeks under the same conditions. Southern and northern analysis indicated in vitro shoot regeneration on the callus derived from the potato explants, which were incubated with Agrobacteria. The regeneration cycle was shortened after the transformatin and finally the transformation efficiency was highly enhanced.

• Journal of Microbiology and Biotechnology
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• v.23 no.10
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• pp.1422-1427
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• 2013

Scutellaria baicalensis (SB), a traditional herb with high pharmacological value, contains more than 10% flavone by weight. To improve the biological activity of flavones in SB, we aimed to enhance the bioconversion of baicalin (BG) to baicalein (B) and wogonoside (WG) to wogonin (W) in SB during fermentation using beta-glucuronidase produced from Lactobacillus brevis RO1. After activation, L. brevis RO1 was cultured in milk containing SB root extract with various carbon or nitrogen sources at $37^{\circ}C$ for 72 h. During fermentation, the growth patterns of L. brevis RO1 and changes in the flavone content were assessed using thin-layer chromatography and high-performance liquid chromatography. After 72 h of fermentation, the concentrations of B and W in the control group increased by only 0.15 and 0.12 mM, respectively, whereas they increased by 0.57 and 0.24 mM in the fish peptone group. The production of B and W was enhanced by the addition of 0.4% fish peptone, which not only improved the growth of L. brevis RO1 (p < 0.001) but also enhanced the bioconversion of flavones. In conclusion, the bioconversion of flavones in SB may provide a potential application for the enhancement of the functional components in SB.

• The Korean Journal of Mycology
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• v.22 no.2
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• pp.138-144
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• 1994

Conditions for isolation of protoplasts from spores and mycelia of Rhizopus nigricans were studied. Larger amount of protoplasts was obtained from swollen spores in liquid medium contained with 5% of 2-deoxy-D-glucose for 4 hours than from mycelia. Enzyme mixture of Novozym 234(2%) and ${\beta}-glucuronidase(5000\;unit/ml)$ was most effective for the isolation of protoplasts from swollen spores and from mycelia. The solution of 0.6 M $MgSO_4$ or mannitol and pH 6.0 showed good results as the osmotic stabilizer and the optimal condition of pH of the enzyme solution for the isolation of protoplast from the swollen spores, respectively. At this condition, $8.0{\times}10^6\;cells/ml$ of protoplasts was obtained from swollen spores by digestion with lytic enzyme mixture for 2 hours.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.46 no.5
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• pp.375-379
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• 2001

Transgenic plants from hypocotyl segments of buckwheat were produced with the Agrobacterium strain LBA4404 harboring the binary vector pBI121 containing chimeric genes of neomycin phosphotransferase II (npt II) and $\beta$-glucuronidase (gus). Two weeks after co-cultivation with Agrobacterium, most of the hypocotyl segments gradually became brown and died on the selection medium containing 100mg/$\ell$ of kanamycin. Plants regenerated from the hypocotyl explants grown on selection medium were GUS-positive in the leaf, stem and vascular tissues by histochemical assay, and varied in gus activity (440-2568 pmol, 4-MU/mg protein) by fluorimetry. The plants showing GUS activity were confirmed of containing GUS and NPT-II genes by polymerase chain reaction (PCR). Within 3 months, transgenic buckwheat plants were able to obtained from the hypocotyl segments.

• The Journal of Internal Korean Medicine
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• v.31 no.4
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• pp.731-751
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• 2010

Objectives : The present study aimed to find out the effect of Sagunja-tang on the prevention and treatment of inflammatory bowel disease using mice with TNBS-induced inflammatory bowel disease. Methods : Mice with TNBS-induced inflammatory bowel disease were medicated with Sagunja-tang, and the weight changes, colon length, lipid peroxidation, and myeloperoxidase activity were observed. Levels of the inflammatory markers interleukin (IL)-$1{\beta}$ and cyclooxygenase-2 (COX-2), its transcription factor activation, phospho-NF-${\kappa}$B (pp65), in the colon by enzyme-linked immunosorbent assay and immunoblot analysis were also measured. Finally, the activation of fecal bacterial enzyme, ${\beta}$-glucuronidase and degradation activation of fecal glycosaminoglycan (GAG) and hyaluronic acid were observed. Results : We found that oral administration of Sagunja-tang inhibited TNBS-induced colon shortening and also inhibited myeloperoxidase activity in the colon of mice as well as IL-$1{\beta}$ and COX-2 expression. Sagunja-tang also inhibited TNBSinduced lipid peroxidation and pp65 activation in the colon of mice. In addition, Sagunja-tang inhibited ${\beta}$-glucuronidase activation and fecal hyaluronic acid degradation activation. Conclusions : It is supposed that Sagunja-tang has a potential therapeutic effect on inflammatory bowel disease through the inhibition of both NF-${\kappa}$B activation and lipid peroxidation, and the improvement of intestinal conditions.

• The Korean Journal of Mycology
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• v.16 no.4
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• pp.214-219
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• 1988

Factors affecting protoplast formation and reversion were investigated in Pleurotus sapidus kalchbr. For release of protoplast, enzyme mixture of Novozyme 234, ${\beta}-D-glucanase$ and ${\beta}-glucuronidase$ was most effective, when mycelium of 0.6 M sucrose solution as osmotic stabilizer without addition of buffer solution. The yield of protoplast was highest with mycelium cultured for 4 days on mushroom complete agar medium at ${30}^{\circ}C$. Protoplasts of Pleurotus sapidus were reverted to normal hyphal growth with maximum reversion frequency of 2% on Mushroom complete agar medium stabilized with 0.6 M sucrose solution and covered by 0.75% agar layer.

• The Journal of Korean Medicine
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• v.29 no.3
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• pp.50-62
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• 2008

Objectives: This study was aimed to find the effect of Samiunkyungtang on inflammation and microflora in an ulcerative colitis animal model. Methods: We established four groups of normal, control, test 1, and test 2 and assigned 6 rats toeach group. The normal group was not treated by any process and fed by normal saline. The control & test groups were provided with 4% dextran sodium sulfate (DSS) treatment for 7 days. Samiunkyungtang extract was orally administered to test groups (test 1=25mg/kg, test 2 100mg/kg) 3 days after DSS treatment for 10 days. After DSS treatment finished, we sacrificed the mice and measured colon length and enzyme activities such as myeloperoxidase (MPO), alkine phosphatase (ALP), ${\beta}$-glucuronidase, ${\beta}$-glucosidase, chondroitinase, and tryptophanase. Results: The colon lengths of test 1 and 2 groups were longer than the control group (p<0.05). Histologically, the crypts and superficial epitheliums of test 1 and 2 groups were regenerated. Goblet cells from all test groups were retrieved. The inflammatory biochemical marker, MPO and ALP activities in all test groups were highly reduced (p<0.01) compared to the control group. The activities of fecal bacterial enzymes in test groups such as ${\beta}$-glucuronidase, ${\beta}$-glucosidase, chondroitinase, and tryptophanase were reduced compared to the control group (p<0.01). Conclusions: As a result of this experiment, Samiunkyungtang is considered to have an inhibitory effect on inflammation and fecal enzyme activity in DSS-induced colitis animal model. Our results indicate that Samiunkyungtang may possess therapeutic effect on the development of DSS-induced colitis.

• Korean Journal of Veterinary Research
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• v.39 no.4
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• pp.765-771
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• 1999

Unlike most Escherichia coli strains, E coli O157 : H7 didn't ferment sorbitol within 24h of incubation and showed a negative reaction for $\beta$-glucuronidase. We developed a new medium for the rapid isolation of E coli O157 : H7 using sorbitol MacConkey agar with cefixime, potassium tellurite and 4-methylumbelliferyl-${\beta}$-D-glucuronide (MUG) as a primary plating medium. The addition of $20{\mu}g/ml$ of vancomycin in enrichment broth for E coli O157 : H7 inhibited lots of Gram positive bacteria. Three strains (10.3%) of 29 non-O157 E coli strains and 3 strains (8.3%) of 36 Salmonella spp were inhibited at the $0.05{\mu}g/ml$ of cefixime and 23 strains (79.3%) of 29 non-O157 E coli strains and 12 strains (33.3%) of 36 Salmonella spp were inhibited at the $2.0{\mu}g/ml$ of potassium tellurite. But none of the E coli O157 : H7 was affected at these concentration. The addition of MUG at $100{\mu}g/ml$ level to sorbitol MacConkey agar with cefixime and potassium tellurite (CTM-SMAC) aided in the rapid isolation of E coli O157 : H7 from samples by checking sorbitol-negative and $\beta$-glucuronidase negative phenotypes simultaneously. In conclusion, inoculation of a positive in the O157 screening test from enrichment broth on CTM-SMAC appeared to be a rapid, cost-effective and sensitive method for the isolation of E coli O157 : H7.

• Korean Journal of Food Science and Technology
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• v.28 no.1
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• pp.53-57
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• 1996

As the production of chonma became increased by the farmer's cultivation, developments of the processed food such as tea using chonma have been actively pursued. In the present study, the components of chonma and its beneficial effects on health using SD (Sprague-Dawley) rat model were analyzed. The contents of moisture, protein, ash, fat and fiber in dried-chonma were 11.8, 7.6, 3.2, 0.5, and 3.9%(w/w), respectively. The contents of calcium, sodium, iron, phosphorus, magnesium and potassium were 121, 83, 6.2, 170, 69 and 1,278 mg%. When chow diets containing 0, 0.15, 1.5 or 5.0% chonma powder were fed to SD rats for 4 weeks, no significant differences were observed in the composition of the large-intestinal flora, ${\beta}-glucuronidase$ level of the large-intestinal contents and the weight gains of rats. The level of ${\beta}-glucosidase$ was higher and the serum cholesterol level was lower in 5.0% chonma group compared with control group. The highest sedative effect was shown in 0.15% chonma group.

• Bulletin of the Korean Chemical Society
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• v.4 no.3
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• pp.139-143
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• 1983

${\beta}$-N-Acetyl-D-glucosaminidase B was highly purified with the following sequence of steps; DEAE-cellulose, CM-cellulose, and Sephadex G-200 gel filtration chromatograpies. The specific activity of the purified ${\beta}$ -N-acetyl-D-glucosaminidase B was 2.2 units/mg protein with 12.9 % yield and 196.2 fold purity. The purified ${\beta}$-N-acetyl-D-glucosaminidase B showed single band on polyacrylamide gel electrophoresis. The final preparation of ${\beta}$ -N-acetyl-D-glucosaminidase B was completely free friom arylsulfatase and ${\beta}$-glucuronidase. ${\beta}$ -N-Acetyl-D-glucosaminidase B had pH optimum of 4.5 in 0.5 M sodium citrate buffer. The molecular weight of ${\beta}$-N-acetyl-D-glucosaminidase B was 133,000 by Sephadex G-200 gel filtration. The Km value of ${\beta}$-N-acetyl-D-glucosaminidase B using p-nitrophenyl-N-acetyl-${\beta}$-D-glucosaminide as substrate was 1.0 mM and $V_{max}$ was 0.014 ${\mu}$ mole/min. ${\beta}$-N-Acetyl-D-glucosaminidase B was stable at $55^{circ}C$ for 70 minutes. The crude ${\beta}$ -N-acetyl-D-glucosamiinidase in 70 % ammonium sulfate retained 93 % activity after 7 months storage at -$55^{circ}C$. Bovine serum albumin, sodium chloride, and phosphate activated ${\beta}$ -N-Acetyl-D-glucosaminidase B. N-Acetyl-D-glucosamine, ${\alpha}$-methyl-D-mannoside, and acetate inhibited ${\beta}$ -N-acetyl-D-glucosaminidase B.