• Korean Journal of Environmental Biology
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• v.25 no.4
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• pp.342-348
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• 2007

In a previous study, a new antibacterial activity was found from Paenibacillus polymyxa DY1 isolated from Korean soil, which showed a good growth inhibition against various multidrug resistant enteric bacteria. Thus the effect of nutritional factors and physicochemical conditions on the growth of P. polymyxa DY1 cells and the production of antibacterial activity were evaluated in this study. For the growth of P. polymyxa DY1 cells the optimum condition reaches by incubating at $35^{\circ}C$ for 48 hours by shaking at 180 rpm in the basal medium containing 0.6% dextrose, 1.4% ammonium phosphate dibasic, 0.9% peptone, 2.4% glutamine, and 2 mM sodium molybdate with initial pH 8.0. A maximum level of antibacterial activity was obtained when P. polymyxa DY1 was incubated at $35^{\circ}C$ for 48 hours by shaking at 180 rpm in the basal medium with initial pH 8.0 containing 1% dextrose, 1.5% ammonium sulfate, 1.1% casamino acid, 1.9% aspartic acid, and 2 mM ferrous sulfate.

• Natural Product Sciences
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• v.11 no.2
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• pp.67-74
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• 2005

The ethanolic extracts of the leaves and bulbs of Allium victorialis var. platyphyllum (Liliaceae) collected from Daegwallyoung (D) and Ullung Island (U) in Korea were obtained using three different extracting methods. The first extracts, DL-1 DB-1, UL-1 and UB-1, were obtained from leaves (L) and bulbs (B) dried at $90^{\circ}C$, respectively, and the second extracts, DL-2, DB-2, UL-2 and UB-2, were obtained by extracting the leaves and bulbs of fresh plant parts. The third extracts DL-3, DB-3, UL-3 and UB-3 were obtained by incubating leaves and bulbs at $36^{\circ}C$. The six extracts obtained from A. victorialis var. platyphyllum at Daegwanllyoung (cultivated site) were orally administered to examine for a possible antihepatotoxic effect in $CCl_4-induced$ rats. DL-1 exhibited the most pronounced effect. The extracts inhibited serum ALT, AST, SDH, ${\gamma}-GT$, ALP and LDH activities elevated by $CCl_4$ injection and attenuated decreased glutathione S-transferase, glutatione reductase and ${\gamma}-glutamylcysteine$ synthetase activities and a decreased hepatic glutathione. However, the extracts obtained from Ullung Is. (native site) were less active than the extracts from Daegwallyoung, suggesting that A. victorialis var. platyphyllum from the cultivated site is more useful for functional food than of native site. These results also suggest that the antihepatotoxic effect is due to a higher content of hepatic glutathione. Gas chromatography of the twelve extracts showed significantly different sulfides, disulfides or trisulfides contents belonging to volatile sulfur substances (VSS). Nine components were identified on the basis of their mass spectra, namely, dimethyl disulfide, dimethyl trisulfide, diallyl disulfide, dipropyl disulfide, allyl methyl sulfide, allyl methyl trisulfide, 2-vinyl-4H-1,3-dithiin, 3,4-dihydro-3-vinyl-1,2-dithiin, and allithiamine. Extract DL-1 had the highest VSS content. Dried plant materials contained larger amounts of the VSSs than other extracts, and the leaves contained larger amount than the bulbs. These results suggest that heat treatment increases the antiheaptotoxic ability of A. victorialis var. platyphyllum by increasing the proportion of VSSs.

• Journal of Life Science
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• v.24 no.12
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• pp.1316-1324
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• 2014

Most beta-glucans obtained from various fruit bodies of mushrooms and mushroom mycelial cultures have high-molecular weight glycoproteins, conjugated with beta-glucans. We report that isoflavone-conjugated glycolproteins (designated as gluvone) were isolated and exhibited stronger anticarcinogenic activities. Agaricus blazei mycelia (ABM) was cultured in a liquid medium containing soybean flakes for 14 days. The liquid culture was autolyzed by incubating at $53^{\circ}C$ (pH 5.5) for 3 h. A crude glycoprotein (CGP) fraction with a cytotoxic effect on a mouse ascite cancer cell line (S-180) and a human breast cancer cell line (MCF-7) was isolated from the autolyzed ABM cultures by 80% ethanol treatment. Gluvone was isolated from the CGP with Sephadex G-75 column chromatography. It exhibited a stronger anticancer effect than CGP against the S-180 cell-induced female ICR mouse ascites carcinogenesis. Gluvone with 9,400 daltons was identified as a glycoprotein conjugated with isoflavone. According to HPLC and GC analysis, in conjunction with $^1H$-NMR spectral analysis, it contained 60% carbohydrates (glucose, fructose, and ribose), 31% protein, and 2% isoflavone (daidzein and genistein), which is a novel material. These results indicate that a strong anticarcinogenic gluvone was isolated from the autolyzed product of a submerged liquid culture of ABM, suggesting that autolysis could be a useful tool to produce antitumor agents.

• Journal of The Korean Society of Grassland and Forage Science
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• v.33 no.2
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• pp.100-104
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• 2013

Zoysiagrass (Zoysia japonica Steud.) is an important forage and turfgrass that spreads by stolons and rhizomes. Zoysiagrass stolon can be used directly for Agrobacterium-mediated genetic transformation by exploiting the potential of direct shoot formation. However, surface sterilization of field-grown stolons is difficult and remains to be explored. We developed an effective surface sterilization and culture method using the stolon explant for infection with Agrobacterium tumefaciens. Among various treatments, sequential disinfection in 30% bleach for 15 min followed by 0.1% mercuric chloride for 25 min resulted in the highest number of clean stolons. The efficacy of mercuric chloride was increased under vacuum conditions by incubating at 800 mbar for 5 min. The inclusion of 2.5 mg/l amphotericin B further prevents fungal growth in in vitro cultures. This protocol would speed up the development of transgenic plants by utilizing field-grown stolon nodes.

• The Korean Journal of Pharmacology
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• v.32 no.2
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• pp.243-257
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• 1996

CCK and cholinergic agonist stimulate enzyme release from the pancreatic acini via G-protein-mediated activation of phospholipase C, In contrast secretin and related peptides increase the level of cAMP and activate cAMP-dependent protein kinase. Camostat, a synthetic protease inhibitor, causes pancreatic hypertrophy and hyperplasia by increasing the CCK release. In this study, the secretagogue-induced changes of intracellular proteins were examined in the dispersed pancreatic acini of rats with or without camostat treatment. Camostat(FOY-305, 200 mg/kg, p.o.) was given for 4 days twice daily and the dispersed acini were prepared at 12 bouts after last treatment. The profiles of Intracellular phosphoproteins were analyzed by two-dimensional gel electrophoresis after incubating the acini with $^{32}P$. The amylase release from the dispersed acini was measured. The pancreatic weight was increased to 126% of control, while amylase activity per mg acinar protein decreased to 41% of control, The maximum response of amylase release from dispersed acini to CCK-8 or carbachol was markedly decreased(65% or 46% of control, respectively). The group of intracellular proteins(24 kD, pI $4.5{\sim}8.5$) was increased in quantity by camostat. CCK-8 or secretin increased phosphorylation of a protein(34 kD, pI 4.7) in camostat-treated as well as control rats. CCK-8 increased tyrosine phosphoryiation in the acini of control rats. However, in camostat-treated rats, the basal level of tyrosine phosphorylation was increased and it was rather decreased by CCK-8. Secretin had no effect on the level of tyrosine phosphorylation in acini. These results indicate that both phospholipase C and adenylate cyclase induce phosphorylation of an intracellular acinar protein(34 kD, pI 4.7) and camostat treatment increases the basal level of tyrosine phosphorylation in acinar cells. And these results suggest that not only serine/threonine protein kinase but also protein tyrosine kinase/phosphatase are involved in the process of CCK receptor mediated stimulation-secrelion coupling.

• Korean Journal of Food Science and Technology
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• v.22 no.6
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• pp.611-617
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• 1990

This study was conducted to breed a high vitamin $B_{12}$ producer by the fusion of protoplasts between Bacillus natto and Bacillus megaterium. Auxotrophic mutants of Bacillus natto SH-34 ($thr^-try^-rif^r$) and Bacillus megaterium BK-13 ($arg^-ade^-lys^-str^r$) which showed high protease activity and production of vitamin $B_{12}$, respectively, were isolated for the fusion experiment. Protoplasts were induced by incubating the cells with lysis solution containing $500{\mu}/ml$ lysozyme, and the ratio of protoplast and regeneration formation were ranged from 99% and 67%, respectively. Fusion frequencies of fusants between Bacillus natto SH-34 and Bacillus megaterium BK-13 were appeared in the ranges of $1.0{\times}10^{-5}$ under the treatment of 30% PEG 6000 containing 3% PVP. The fusant, MNF-72 showed the highest product yield of $7.85{\mu}g/g-cell\;vitamin\;B_{12}$ in production medium. For the improvement of productivity, the immobilization of fusants with sodium alginate was carried out. In batch and continuous fermentation systems, the productivity were determined to be $0.58{\mu}g/ml.hr\;and\;0.80{\mu}g/ml.hr\;vitamin\;B_{12}$ under optimum condition, respectivity.

• Journal of Oral Medicine and Pain
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• v.26 no.1
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• pp.39-50
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• 2001

In order to investigate the effect of Transforming growth factor ${\beta}1$(below TGF-${\beta}1$) and osteogenic protein-1(below Op-1) onto the myogenic differentiation, C2C12 satellite myoblastic cell line was cultured and treated with both growth factors. At first morphological changes with microscopical examination were examined, and isolated total RNA to analyse mRNA expression of bone marker proteins, muscle regulatory proteins, TGF-${\beta}$ receptor and their ligands by Northern blot analysis. And cellular proliferative inducibility of both growth factors was also tested to C2C12 cells. Incubating the cell with $5ng/m{\ell}$ of TGF-${\beta}1$ until 4 days almost inhibited multinucleated myotube formation expressing muscular regulatory proteins, and induced decreasing Id proteins. However, no osteoblastic phenotypes was induced by TGF-${\beta}1$ in C2C12 cells. The mRNA expression of TGF-${\beta}$ receptors with TGF-${\beta}1$ was conversed after 48 hours cultured. Type I TGF-${\beta}$ receptor was seemed to play a role in negative signalling for inhibition of myogenic differentiation. OP-1 dose dependently induced ALP activity, osteopontine production and bone sialoprotein production at concentrations above $100ng/m{\ell}$ and osteocalcin production at concentrations above $300ng/m{\ell}$. The concentration of OP-1 required to induce these osteoblastic phenotypes was the same as that required to almost completely inhibit myotube formation. Incubation with above $100ng/m{\ell}$ OP-1 suppressed the expression of mRNA for muscular egulatory proteins from 2 days after incubation. Expression of Id-1, 2, 3 mRNA were stimulated by OP-1 at concentration above $300ng/m{\ell}$. When C2C12 cells were treated with both growth factors, TGF-${\beta}1$ potentiated the inhibitory effect of OP-1 on myotube formation and expression of mRNA for myogenin at 12 days. And TGF-${\beta}1$ reduced osteocalcin and bone sialoprotein production induced by OP-1 at 12 days in C2C12 cells. Both growth factor had no mitogenic effect. These results indicate that OP-1 converts the differentiation pathway of C2C12 myoblasts into that of osteoblastic lineage cells and it's not heritable, but TGF-${\beta}1$ does not and has reversible inhibitory activity on the myogenic differentiation. TGF-${\beta}1$ and OP-1 play a role in myogenic differentiation via different mechanism between them.

• Journal of Digital Convergence
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• v.15 no.1
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• pp.347-353
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• 2017

This study aims to figure out how green tea leaves, mulberry leaves, and mate leaves that are often taken around in drinking tea influence on S. mutans known as dental caries causative bacteria. After vaccinating 1% of S. mutans on the extract added badge and incubating 6 hours and 10 hours in $37^{\circ}C$, it calculated absorbance and bacterial colony number. Extract of green tea leaves and mate leaves were added with 0, 0.5, 1.0, 2.0, and 4.0% of concentration. As a result of growth suppressive effect of S. mutans, it may be observed that colony number significantly decreases in higher concentration. When it calculated bacterial colony number with adding 2% extract after 10 hours, 99.0% for green tea leaves, 97.1% for mulberry leaves, and 89.6% for mate leaves appeared.

• Korean Journal of Microbiology
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• v.46 no.2
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• pp.127-133
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• 2010

The purpose of this work was to investigate the antibacterial activity derived from a lactic acid bacterium, Lactococcus lactis HK-9, isolated from the feces of a 2-day newborn infant. We characterized the physiological and biochemical properties of this strain. Both the BIOLOG system and phylogenetic analysis using 16S rRNA sequencing were utilized for identification, and the strain was assigned to the Lactococcus lactis species, designated as L. lactis HK-9, and registered in GenBank as [GU936712]. We monitored growth rate, production of lactic acid and acetic acid as metabolites, and pH during growth. The maximum concentrations of lactic acid and acetic acid reached 495.6 mM and 104.3 mM, respectively, and the initial pH of the cultures decreased from 7.0 to 4.1 after incubating for 60 h. HPLC was used to confirm the production of lactic acid and acetic acid. Significant antibacterial activity of the concentrated supernatant was demonstrated against Gram-positive (e.g., Staphylococcus aureus, Enterococcus faecalis, Listeria monocytogenes, MRSA) and Gram-negative (e.g., Escherichia coli, Salmonella enteritidis, Pseudomonas aeruginosa, Klebsiella pneumoniae, Shigella sonnei) bacteria by the plate diffusion method. The antibacterial activity was sensitive to protease, and the molecular weight of the presumed bacteriocin molecule was estimated to be about 4 kDa by tricine-SDS-PAGE.

• Food Science of Animal Resources
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• v.22 no.3
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• pp.274-280
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• 2002

This study was conducted to investigate inhibitory effects of K-casein, GMP and sialic acid addition on the infection of MA-104 cells by 597(Korean native cattle rotavirus) and JBR(Jeju island bovine rotavirus). MA-104 cells on incomplete Ml99 were infected with domestically separated 597 and ma activated by incubating at 37$\^{C}$ for 6 days, and analyzed for the titer of rotavirus. K-casein, GMP and sialic acid added MA-104 culture infected by activated S97 and nan were incubated for Is hours and stained by the AEC stainning method. The number of infected cells were counted on microscope. The titer of S97 and JBR was 2.5$\times$107 and 2.0$\times$106 PFU/ml, respectively. The inhibition level against cell infection by 597 was 97.4% far 2000UH of K-casein and 97.44% for 2000UM of GMP. The inhibition level against cell infection by JBR was 99.52% for 2000$\mu$M of $\kappa$-casein and 99.78% for 2000$\mu$M of GMP. The inhibition level against cell infection by 597 and JBR was 3.85 and 3.63% for 2000$\mu$M of sialic acid, respectively. The high inhibitory effects (over 97%) of K-casein and CMP against infection of U-1(14 cells with 597 and mR indicated great potentials for the use of K-casein and GMP in the treatment of calf or infant caused by rotavirus.