• Journal of the Korean Society of Food Science and Nutrition
• /
• v.40 no.6
• /
• pp.903-911
• /
• 2011

In this study, Bacillus polyfermenticus CJ6 harboring antibacterial activity was isolated from meju. The antibacterial activity of Bacillus polyfermenticus CJ6 was stable in the pH range of 3.0~9.0, but it disappeared after culture at $70^{\circ}C$ for 24 hr. Antibacterial activity was inactivated by proteinase K, protease, and ${\alpha}$-chymotrypsin, indicating its proteinaceous nature. The growth inhibitory effects of B. polyfermenticus CJ6 culture on food-borne pathogens such as Staphylococcus aureus, Salmonella Typhi, Listeria monocytogenes, and Escherichia coli O157:H7 were examined in this study. Approximately 6~6.2 log CFU/mL of each pathogen was co-cultured with B. polyfermenticus CJ6 in a 50 mL culture volume for 24 hr. Growth of S. aureus and L. monocytogenes was completely inhibited after 3 hr of incubation. Growth of S. Typhi and E. coli O157:H7 was also completely inhibited after 6 hr of incubation. The antibacterial compounds from B. polyfermenticus CJ6 were purified by solid phase extraction (C18 Sep-pak cartridge), recycling preparative HPLC, and analytical HPLC. Ultra-high performance liquid chromatography and electrospray ionization tandem mass spectrometry analysis were used to identify the purified antibacterial compounds, which were confirmed to be five peptides (757.4153 Da, 750.3444 Da, 1024.5282 Da, 1123.6083 Da, and 1617.8170 Da).

• Korean Journal of Microbiology
• /
• v.46 no.2
• /
• pp.192-199
• /
• 2010

The alone and combined effects of bacteriocin produced from Enterococcus faecalis MJ-213 and potassium sorbate against the food-borne pathogenic bacteria were studied. Bacteriocin minimal inhibitory concentration (MIC) values for Staphylococcus aureus ATCC 6538 and Salmonella enteritidis ATCC 13076 were 50 and 100 ${\mu}g$/ml, respectively. Bacteriocin (100 ${\mu}g$/ml) alone was active against S. aureus and S. enteritidis, but it was lower in antimicrobial effectiveness than the combination of bacteriocin (100 ${\mu}g$/ml) with potassium sorbate (100 ${\mu}g$/ml), which reduced initial counts (6 log cycle) of S. aureus and S. enteritidis by 1 and 3 log cycle, respectively. The bactericidal activity of bacteriocin of E. faecalis MJ-213 heated at $100^{\circ}C$ for 30 min or $121^{\circ}C$ for 15 min was markedly decreased as compared with the control. Moreover, the activity of bacteriocin was completely abolished by pepsin or protease II, but not affected by ${\alpha}$-amylase or lipase. The activity of bacteriocin adjusted to pH 6.0-8.0 showed almost the same inhibition ratio compared with the bacteriocin unadjusted pH, and though the inhibition ratio against pathogenic bacteria was reduced than the control, the bacteriocin was stable at pH 4.0 or 10.0, relatively. Furthermore, the combined treatment of bacteriocin and potassium sorbate than the alone treatment of bacteriocin significantly decreased (p<0.05) the viable cell counts of S. aureus or S. enteritidis inoculated on grind beef during storage at $4^{\circ}C$.

• Journal of dental hygiene science
• /
• v.6 no.1
• /
• pp.1-9
• /
• 2006

Cathepsin K (cat K) is the major cysteine protease expressed in osteoclasts and was thought to play a key role in matrix degradation during bone resorption. It was shown that the intracellular maturation of cat K was prevented by the cAMP antagonist, Rp-cAMP, and the protein kinase A (PKA) inhibitors of KT5720 and H89. In contrast, forskolin, a adenylate cyclase agonist, rather induced Cat K processing and maturation in osteoclasts. Furthermore, to determine whether cat K processing and maturation signaling involves protein kinase C (PKC), mouse total bone cells were treated with calphostin C, a specific inhibitor of PKC, however, no effect was observed, indicating that calphostin C did not affect to osteoclast-mediated cat K processing and maturation. Thus, it is indicated that the cAMP-PKA signaling pathway regulates cat K maturation in osteoclasts. Since secreted proenzymes have the potential to reenter the cell via M6P receptor, to prevent this possibility, it was tested cAMP antagonist Rp-cAMP and the PKA inhibitors KT5720 and H89 in the absence or presence of M6P. Inhibition of cat K processing by Rp-cAMP, KT5720, or H89 was observed in a dose-dependent manner. Furthermore, the addition of M6P resulted in enhanced potency of Rp-cAMP, KT5720 and H89. These dose-dependently inhibited in vitro bone resorption with a potency similar to that observed for inhibition of cat K processing.

• Journal of Life Science
• /
• v.7 no.4
• /
• pp.392-401
• /
• 1997

The poliovirus is a small, and non-enveloped virus. The RNA genome of poliovirus is continuous, linear, and has a single open reading frame. This polyprotein precursor is cleaved proteolytically to yield mature products. Most of the cleavages occur by viral protease. The mature proteins derived from the P1 polyprotein precursor are the structural components of the viral capsid. The initial cleavage by 2A protease is indirectly involved in the cleavage of a cellular protein p220, a subunit of the eukaryotic translation initiation factor 4F. This cleavage leads to the shut-off of cap-dependent host cell translation, and allows poliovirus to utilize the host cell machinery exclusively for translation its own RNA, which is initiated by internal ribosome entry via a cap-independent mechanism. The functional role of the 2B, 2C and 2BC proteins are not much known. 2B, 2C, 2BC and 3CD proteins are involved in the replication complex of virus induced vesicles. All newly synthesized viral RNAs are linked with VPg. VPg is a 22 amino acid polypeptide which is derived from 3AB. The 3C and 3CD are protease and process most of the cleavage sites of the polyprotein precursor. The 3C protein is also involved in inhibition of RNA polymerase II and III mediated transcription by converting host transcription factor to an inactive form. The 3D is the RNA dependent RNA polymerase. It is known that poliovirus replication follows the general pattern of positive strand RNA virus. Plus strand RNA is transcribed into complementary minus strand RNA that, in turn, is transcribed for the synthesis of plus strand RNA is transcribed into complementary minus strand RNA that, in turn, is transcribed for the synthesis of plus strand RNA strands. Poliovirus RNA synthesis occurs in a membranous environment but how the template RNA and proteins required for RNA replication assemble in the membrane is not much known. The RNA requirements for the encapsidation of the poliovirus genome (packaging signal) are totally unknown. The poliovirus infection cycle lasts approximately 6 hours.

• Journal of Mushroom
• /
• v.12 no.3
• /
• pp.201-208
• /
• 2014

The separation of the bacteria inhibiting Trichoderma sp. mold, the strain causing blue mold disease that occurs frequently when cultivating mushroom while carrying out the efficient fermentation of mushroom medium, from the growth was done. In about 200 strains isolated primarily from fungus garden samples, 6 strains were secondly isolated, which had fast growth rates and a clear zone on the plate medium of SM, AM, and CM. Among the 6 strains isolated, the C-1 strain showed high enzymatic activity of cellulase, amylase, and protease, and strong antibacterial activity for the T. virens and T. harzianum, selected finally. The selected C-1 strain was identified as Paenibacillus polymyxaby the result of the identification by Bergey's Manual of Systematic Bacteriology and the analysis of the nucleotide sequence of 16S rRNA, and named as P. polymyxa CK-1. In reviewing the growth conditions of the P. polymyxa CK-1 strain, the optimum cultivation temperature was $45^{\circ}C$, and the optimum pH for growth was in the range of 6.0~7.0. Appropriate incubation time of P. polymyxa CK-1 for the growth inhibition of the fungus T. virens and T. harzianum was 22 to 36 hours. And the fungal growth was not observed, even when leaving two molds inoculated on each petri dishes, which were treated with 24 hour culture solution of P. polymyxa CK-1 strain for 10 days. As a result of studying the thermal stability of the antagonists produced by the P. polymyxa CK-1 strain, no mycelial growth of the two fungi was observed in the test group treated for 20 minutes at $60^{\circ}C$ and $100^{\circ}C$, but mycelial growth was slightly observed in the test group treated for 20 minutes at $121^{\circ}C$. As aresult of reviewing the impact of the P. polymyxa CK-1 culture medium on mushroom mycelial growth, it showed no effect on a variety of mushroom mycelial growth including enoki mushroom and shiitake mushroom.

• Journal of the Society of Cosmetic Scientists of Korea
• /
• v.45 no.4
• /
• pp.389-397
• /
• 2019

Lysosomes are cellular organelles involved in energy metabolism and intracellular digestion in eukaryotic cells, including protease, nuclease, glycosidase, lipase, and phosphatase. Our previous studies have confirmed that egg white lysosomes had melanin decolorization and reduction activity. However, there have been few studies on skin barrier and skin regeneration as well as inhibition of melanin production by egg white lysosomes on B16F10 melanocyte cell line. In this study, we attempted to identify the effect of lysosome-related organelle extract (LOE) extracted from egg white on the melanin content change and skin barrier enhancement in cells. First, cytotoxicity evaluation was performed on B16F10 melanocyte cell line to confirm the whitening efficacy of LOE. Cytotoxicity by LOE was not observed at 20 mg/mL concentration, but cytotoxicity was observed at 40 mg/mL, and the maximum concentration value was set to 20 mg/mL in all subsequent experiments. LOE samples of 5, 10, 20 mg/mL inhibited melanin production by 61.5 ± 4.0%, 61.4 ± 7.3%, 58.3 ± 8.3%, respectivly, compared to α-MSH, a negative control in melanin contents assay. MITF mRNA expression was reduced by about 39.7 ± 3.2% compared to the α-MSH treatment group. TEER assay using HaCaT showed that LOE increased TEER resistance in a dose-dependent manner, indicating that LOE is involved in strengthening the skin barrier. LOE also increased the TEER resistance under TNF-α treatment. Skin barrier was normally restored by LOE even under the condition of inflammation. LOE had a positive effect on cell division and cell migration promotion, confirmed by the observing the effect of promoting cell migration by LOE through cell migration assay. Taken together, we expect that LOE can be developed as a cosmetic material to enhance has effects on skin regeneration and skin barrier strengthening as well as whitening function if enzyme stabilization and formulation technology are combined.

• Korean Journal of Food Science and Technology
• /
• v.44 no.6
• /
• pp.743-749
• /
• 2012

Black soybean paste (BSP) is a bealmijang manufactured with barley flour and black soybeans. This study investigated the quality characteristics of BSP prepared with Bacillus subtilis HJ18-4 (HJ18-4). ${\alpha}$-Amylase and protease activities of BSP rapidly increased on day 6 and day 9, respectively, ($41.45{\pm}0.3-42.72{\pm}0.16$, $21.84{\pm}0.24-23.11{\pm}0.35unit/g$) after which the activities were constant or decreased slightly. Amino type nitrogen contents increased until day 23 ($173.78{\pm}3.51-195.63{\pm}1.51mg\;%$), whereas ammonia type nitrogen sharply decreased on day 9. On day 30, lactic acid bacteria counts were higher compared with the initial count ($7.91{\pm}0.05-8.17{\pm}0.02log\;CFU/g$). Meanwhile, B. cereus and total aerobic counts in HJ18-4 added BSP were lower than that in the control. These results implied that BSP could be edible after 30 days of fermentation, and the addition of HJ18-4 in meju could contribute to the safety of the black soybean paste by the inhibition of B. cereus growth.

• Korean Journal of Soil Science and Fertilizer
• /
• v.36 no.3
• /
• pp.134-144
• /
• 2003

A rhizobacterium Pseudomonas cholororaphis O6 produced several secondary metabolites, such as phenazines, protease, and HCN that may be involved in inhibition of the growth of phytopathogenic fungi. In field study, P. chlororaphis O6 treatment on wheat seed suppressed root rot disease caused by Fusarium culmorum. The major organic acids of cucumber root exudates were fumaric acid, malic acid, benzoic acid, and succinic acid. Glucose and fructose were major monosaccharides in cucumber root exudates. The total amount of organic acids was ten times higher than that of the sugars. P. chlororaphis O6 grew well on cucumber root exudates. The dctA gene of P. chlororaphis O6 consisted of a 1,335 bp open reading frame with a deduced amino acid sequence of 444 residues, corresponding to a molecular size of about 47 kD and pI 8.2. The deduced dctA sequence has ten putative transmembrane domains, as expected of a membrane-embedded protein. Our results indicated that organic acids in cucumber root exudates may play an important role in providing nutrient source for root colonization of biological control bacteria, and the dctA gene of P. chlororaphis O6 may be an important bacterial trait that is involved in utilization of root exudates.

• Journal of Life Science
• /
• v.29 no.3
• /
• pp.295-302
• /
• 2019

Liquiritigenin (LG) is a chiral flavonoid isolated from the roots of licorice. It exhibits multiple biological activities including anti-oxidant, anti-cancer, and anti-inflammatory effects. In particular though, the anti-cancer activity of LG in oral squamous cell carcinoma has yet to be elucidated, and LG-induced apoptosis in oral squamous cell carcinoma remains poorly understood. In the present study, we tested the role of LG in inducing apoptosis in oral squamous cell carcinoma cells. LG treatment of HN22 cells resulted in a dose-dependent inhibition of cell viability as detected by a 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide assay. The induction of apoptosis in terms of Annexin V/7-Aminoactinomycin D staining, sub-G1 population, and multi-caspase activity were assessed with a $Muse^{TM}$ Cell Analyzer. Flow cytometric analysis revealed that LG treatment resulted in G2/M arrest in cell cycle progression and downregulation of cyclin B1 and CDC2 expression in a concentration-dependent manner. It also resulted in significant upregulation of p27. In addition, LG was seen to trigger the generation of reactive oxygen species and induce CCAAT/enhancer-binding protein homologous protein and 78-kDa glucose-regulated protein in concentration-dependent upregulation. The LG treatment of HN22 cells led to a loss of mitochondrial membrane potential (${\Delta}{\Psi}m$); it also reduced the levels of anti-apoptotic protein and increased the expression of apoptotic protease activating factor-1, cleaved poly (ADP-ribose)polymerase and Bax. Overall, our results indicate that the pro-apoptotic effects of LG in HN22 cells depend on the activation of both intrinsic and extrinsic signaling pathways. Thus, our results suggest that LG constitutes a natural compound with a potential role as an anti-tumor agent in oral squamous cell carcinoma.

• Journal of Acupuncture Research
• /
• v.21 no.2
• /
• pp.41-55
• /
• 2004

Objective : To investigate the possible molecular mechanism (s) of melittin as a candidate of anti-cancer drug, we examined the effects of the compound on the growth of human lung carcinoma cell line A549. Methods : Growth inhibitory study, flow cytometry analysis, SDS-polyacrylamide gel electrophoresis and Western blot analysis, RT-PCR and in vitro caspases activity assay were performed. Results : Melittin treatment declined the cell viability of A549 cells in a concentration-dependent manner, which was associated with induction of apoptotic cell death. Melittin treatment down-regulated the levels of Bcl-XS/L mRNA and protein expression of A549 cells, an anti-apoptotic gene, however, the those of Bax, a pro-apoptotic gene, were up-regulated. Melittin induced the proteolytic cleavage and activation of caspase-3 and caspase-9 protease in a dose-dependent manner without alteration of inhibitor of apoptosis proteins family and Akt expression. Western blot analysis and RT-PCR data revealed that the levels of tumor suppressor p53 and cyclin-dependent kinase inhibitor p21 were also remained unchanged. Conclusions : Taken together, these findings suggest that melittin-induced inhibition of human lung cancer cell growth is associated with the induction of apoptotic cell death via regulation of several major growth regulatory gene products, and melittin may have therapeutic potential in human lung cancer.