• The Journal of the Korean Society for Microbiology
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• v.35 no.4
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• pp.317-324
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• 2000

Candida albicans is one of the most frequently isolated fungal pathogens in human. Recently, the prevalence of candida infection has markedly increased, partially due to the increase of immunocompromised hosts. Proposed virulence factors of the pathogenic Candida are the ability to form hyphae to adhere to epithelial cell surfaces, and to secrete acid proteinases and phospholipases. We measured the relative cell surface hydrophobicity (CSH) and the ability of proteinase production (PROT), phospholipase production (PLase), adherence to host epithelium (ADH), and hyphal transition (Germ). The relative risk of virulence factors was analyzed by lethality test in murine model of hematogeneously disseminated candidal infection. According to Cox's proportional hazard analysis, the statistically significant virulence factors were PROT, ADH, and CSH. PROT was the highest risk factor of them. To evaluate the applicability for the diagnosis and treatment of Candidiasis, we examined the protective effect of the active and passive immunizations with the materials purified from virulence factors and antibodies to them in Candia-infected mice model. The mean survival times of active and passive immunized groups were slightly longer than those of non-immunized groups.

• Archives of Pharmacal Research
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• v.28 no.7
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• pp.816-822
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• 2005

Catharsius protease-2 (CPM-2) was isolated from the body of dung beetles, Catharsius molossus, using a three step purification process (ammonium sulfate fractionation, gel filtration on Bio-Gel P-60, and affinity chromatography on DEAE Affi-Gel blue). The purified CPM-2, having a molecular weight of 24 kDa, was assessed homogeneously by SDS-polyacrylamide gel electrophoresis. The N-terminal amino acid sequence of CPM-2 was composed of X Val Gin Asp Phe Val Glu Glu lie Leu. CPM-2 was inactivated by $Cu^{2+}\;and\;Zn^{2+}$ and strongly inhibited by typical serine proteinase inhibitors such as TLCK, soybean trypsin inhibitor, aprotinin, benzamidine, and ${\alpha}_1$-antitrypsin. However, EDTA, EGTA, cysteine, $\beta$-mercaptoethanol, E64, and elastatinal had little effect on enzyme activity. In addition, antiplasmin and antithrombin III were not sensitive to CPM-2. Based on the results of a fibrinolytic activity test, CPM-2 readily cleaved $A{\alpha}-$ and $B{\beta}$-chains of fibrinogen and fibrin, and y-chain of fibrinogen more slowly. The nonspecific action of the enzyme resulted in extensive hydrolysis, releasing a variety of fibrinopeptides of fibrinogen and fibrin. Polyclonal antibodies of CPM-2 were reactive to the native form of antigen. The ELISA was applied to detect quantities, in nanograms, of the antigen in CPM-2 protein.

• Korean Journal of Microbiology
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• v.8 no.3
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• pp.107-115
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• 1970

In order to brew Takju, Korean flour wine, it requires three necessary steps for specific brewage. One is primary brewing process and another main brewing process to sacharify and perform alcoholic fermentation. After previous two brewing passing, the mash of main brewing process mixed with 1 volume of water is commercial Takju. Three samples were obtained from three breweries and incubated at $28{\pm}1^{\circ}C$for the alcoholic fermentation. All the samples were analyzed for observation of the changes of various biochemical constituents which were contained in the mashes of two brewing processes and in Takju. The starch contents of the suspensions in the primary and main mashes, and in Takju were 28.08, 25.92, 3.83%, and decreased considerably within 36-48 hrs and thereafter slowly. The amounts of water soluble carbohydrates and reducing sugars in three steps had a tendency of decreasing within 48 hours. The initial numbers of yeasts per 1ml in the suspensions were $1.74{\times}10^8$, $1.65{\times}10^8$, $0.66{\times}10^8$, appeared the highest at 72 hours (12.66%) in main process, and in the case of Takju kept increasing untill 60 hours. The dextrin contents in the mashe of primary process through the long period were 0.34 - 0.68%, in the main one were 0.12 - 0.32%, concerning with the amylase activities which were stronger in the mash of the primary process than in that of the main. The contents of methanol and cellulose in the mashes of two processes were 3.40-0.68%, in the main one were 0.12-0.32%, concerning with the amylase activities which were stronger in the mash of the primary process than in that of the main. The contents of methanol and cellulose in the mashes of two processes were 3.40-5.98mg/ml, 0.43-0.48% during the fermentation. The consumption of crude protein the mashes of Takju revealed with time, on the contrary, the production of free amino acids and oligo-peptides were performed, depending upon the affection of proteinase. The amount of tannic acid in Takju was 0.0073-0.0098mg/ml and organic acids of these three groups increased with time and hydrogen ion concentration was 3.28-4.43.

• Journal of Microbiology and Biotechnology
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• v.32 no.11
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• pp.1462-1470
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• 2022

Natural antimicrobial substances are needed as alternatives to synthetic antimicrobials to protect against foodborne pathogens. In this study, a bacteriocin-producing bacterium, Bacillus subtilis HD15, was isolated from doenjang, a traditional Korean fermented soybean paste. We sequenced the complete genome of B. subtilis HD15. This genome size was 4,173,431 bp with a G + C content of of 43.58%, 4,305 genes, and 4,222 protein-coding genes with predicted functions, including a subtilosin A gene cluster. The bacteriocin was purified by ammonium sulfate precipitation, Diethylaminoethanol-Sepharose chromatography, and Sephacryl gel filtration, with 12.4-fold purification and 26.2% yield, respectively. The purified protein had a molecular weight of 3.6 kDa. The N-terminal amino acid sequence showed the highest similarity to Bacillus subtilis 168 subtilosin A (78%) but only 68% similarity to B. tequilensis subtilosin proteins, indicating that the antimicrobial substance isolated from B. subtilis HD15 is a novel bacteriocin related to subtilosin A. The purified protein from B. subtilis HD15 exhibited high antimicrobial activity against Listeria monocytogenes and Bacillus cereus. It showed stable activity in the range 0-70℃ and pH 2-10 and was completely inhibited by protease, proteinase K, and pronase E treatment, suggesting that it is a proteinaceous substance. These findings support the potential industrial applications of the novel bacteriocin purified from B. subtilis HD15.

• Development and Reproduction
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• v.7 no.2
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• pp.113-118
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• 2003

Follicular fluid(FF) of mammalian Graafian follicles contains various kinds of proteins and proteinases that are believed to play important roles during follicular growth oocyte maturation and ovulation of mature oocytes. Previous studies of human FF(hFF) demonstrated the presence of many serine/threonine proteinases and matrix metalloproteinases such as gelatinases, however, little is known about the caseinases. Present study was aimed to examine the presence and the property of caseinolytic enzyme in hFF. Using casein zymographic method, it was found that hFF, human adult serum and cord serum exhibited one intense 80 kDa and another weak 78 kDa bands having caseinolytic activity. When inhibitors were added to the zymographic substrate buffer, caseinolytic activity of both 80 kDa and 78 kDa proteins were inhibited by othylenediarnine tetraacetic acid(EDTA) or soybean trypsin inhibitor(SBTI), but not by E-64, phenylmethylsulfonyl fluoride(PMSF) or 1,10-phenanthroline. Thus both enzymes appear to belong to a family of trypsin-like enzyme. Addition of EDTA to the zymographic substrate buffer almost abolished the caseinolytic activity of both enzymes. However, further addition of a divalent metal ion such as CaC $l_2$, MgC $l_2$, MnC $l_2$ or ZnC $l_2$ to the same buffer fully restored the enzyme activity at 5 mM concentration despite the presence of EDTA. Based upon these observations, 80 kDa and 78 kDa caseinolytic enzymes are present in human follicular fluid and they appear to be trypsin-like enzymes of which caseinolytic activity needs the presence of $Ca^{++}$, aM $g^{++}$, M $n^{++}$ or Z $n^{++}$././././.

• Korean Journal of Poultry Science
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• v.30 no.1
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• pp.11-16
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• 2003

This study was conducted to investigate chemical and sensory characteristics of boiled soup extracted from crossbred ogol chicken as affected by the level of Flavourzyme produced from Aspergillus oryzae and composed with endo-proteinase(674U/g) and exoproteinase(8,053U/g). It was hydrolyzed by different concentrations of the protease enzyme (Flavourzyme)(0%(Control), 0.01％(T$_2$), 0.1%(T$_3$) and 0.5%(T$_4$)) at 45$^{\circ}C$ for 4hrs. Manufacture of the extract was performed by boiling treated meats with medicinal herbs(Sipchun daebo) at a higher pressure condition. Minerals, free amino acid content, sensory properties of the extract were as follows. The sodium contents were increased as the treatment levels of enzyme increased. The iron contents were lowest when the enzyme treated by 0.5% level, however there were not significantly different among the treatments. The copper and mangan contents had no significantly different among the enzyme treatment levels. Higher contents of fee amino acid were observed as the amount of the protease increased with the treatment of higher than 0.1％ enzyme, no significant effect was observed. In sensory properties, the extract manufactured by addition of 0.01∼0.1% of Flavourzyme resulted in a similar or better appearance, flavor, taste and overall palatabilitycompared to control(no enzyme treatment). However, the extract manufactured with 0.5% of Flavourzyme resulted in lower scores in appearance, flavor, taste and overall palatability than the control. In addition, this product showed more off-flavor than control.

• Journal of Life Science
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• v.14 no.4
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• pp.683-688
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• 2004

A bacteriocin-producing lactic acid bacteria was isolated from Kimchi on MRS selective media with the use of Lactobacillus delbrueckii subsp. delbrueckii as an indicator strain. The strain YH-10 was identified as Lactococcus lactis subsp. lactis through the API test. The crude bacteriocin (freeze-dried 50% ammonium sulfate precipitate of culture supernatant) produced by the strain was named as lacticin YH-10. Lacticin YH-10 showed the growth inhibitory activity against Gram positive pathogenic bacteria and other lactic acid bacteria. The bacteriocin was inactivated by proteases such as protamex and aroase AP-10 and partially inactivated by amylase, proteinase K, trypsin, and papain. The lacticin YH-10 remained its activity with the treatment of heat at 10$0^{\circ}C$ for 60 min or the changes of pH 2 to 11. However, the activity was lost at high pH combined with the exposure to 10$0^{\circ}C$. The bacteriocin production of the strain was started in the exponential phase and stopped in the stationary phase. The approximate molecular mass of the bacteriocin produced by the strain was approximate 14 kDa in the analysis on SDS-PAGE.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.44 no.1
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• pp.49-57
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• 2018

In this study, we investigated the anti-aging effect of Rosa damascena extract containing low molecular glycoprotein (RELG) converted from the high molecular glycoprotein by bioconversion. Free radical scavenging activities were performed by 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. Antioxidant activities ($IC_{50}$) of RELG and the positive control ascorbic acid were $22.6{\mu}g/mL$ and $21.1{\mu}g/mL$, respectively. For skin cells, $15{\mu}g/mL$ RELG showed 28% antioxidant activity by inhibiting the production of active oxygen species induced by ultraviolet ray and hydrogen peroxide. $15{\mu}g/mL$ RELG prevented 10% the cell death caused by stress in human hair follicle dermal papilla cells (HDPC) and reduced 90% the production of active oxygen species. In addition, the glycoprotein showed not only anti-wrinkle effect but also moisturizing effect by 48% inhibition of matrix metallo proteinase-1 (MMP-1) production by ultraviolet stress and $10{\mu}g/mL$ RELG enhanced 10% neutral lipid synthesis with 44% aquaporin 3 (AQP3) expression, which is moisture factor. In conclusion, the RELG can be used as an anti-aging cosmetic material.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.38 no.4
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• pp.350-359
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• 1993

The effect of gibberellic acid ($GA_3) and abscisic acid (ABA) on KCl-enhanced proteolysis of senescing leaves of rice(Oryza sativa L. cv. Chilsung) was studied. Emphasis was given to their effects on KCI-enhanced efflux of amino acids and proteinase activity. When treated singly, $GA_3 affected leaf proteolysis little, while ABA increased proteolysis, the rate of amino acid efflux, and ribulose -1,5 -bisphosphate carboxylase / oxygenase (Rubisco)-degrading endoproteinase activity. An additive increase in all three parameters mentioned above was observed when leaves were treated with ABA and KCl. No such an additive effect was found when $GA_3 was treated with KCl. Both $GA_3 and ABA helped to alleviate the KCI-suppressed activity of Rubisco-degrading exoproteinases. The additive increase in proteolysis of rice leaves in the presence of both ABA and KCl could thus be ascribed to a further increase in the efflux of protein hydrolyzates and Rubisco-degrading endoproteinase activity. An increase in proteolysis was accompanied by a decrease in water absorption, and the combined treatment of ABA with KCl resulted in a further reduction of water absorption.