• Journal of Physiology & Pathology in Korean Medicine
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• v.23 no.6
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• pp.1444-1448
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• 2009

Potentilla chinensis Ser. (Rosaceae) has long been used for a remedy of diarrhea and inflammation in Korea. In this study, the anti-inflammatory effects of the Potentillae chinensis Herba water extract (PCX) was investigated in proteinase-activated receptor-2 (PAR2)-mediated rat paw edema. Paw edema was induced by injection of trypsin or trans-cinnamoyl-LIGRLO-$NH_2$ (tc-$NH_2$) into the hind paw of rats. PCX (10, 50, 100 and 200 mg/kg) was orally administered 1 h before the induction of inflammation. At doses of 50, 100 and 200 mg/kg, PCX showed significant inhibition on both change in paw volume and vascular permeability. PCX (100 mg/kg) significantly inhibited PAR2 agonists-induced myeloperoxidase (MPO) activity in paw tissue. These results indicate that PCX has an anti-inflammatory action in PAR2-mediated paw edema.

• Archives of Pharmacal Research
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• v.26 no.9
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• pp.731-734
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• 2003

Five known kaurane type diterpenoids, 16$\alpha$H, 17-isovaleryloxy-ent-kauran-19-oic acid (1), 16$\alpha$-hydroxy-17-isovaleryloxy-ent-kauran-19-oic acid (2), paniculoside-IV (3), 16$\alpha$-hydroxy-ent-kauran-19-oic acid (4), and ent-kaur-16-en-19-oic acid (5) were isolated from the root of Acanthopanax koreanum by repeated column chromatography and reversed phase preparative HPLC. The structures of these compounds were established from physicochemical and spectral data. Among the isolated compounds 16$\alpha$H, 17-isovaleryloxy-ent-kauran-19-oic acid (1) showed potent inhibitory activity ($IC_50$ value, 16.2 $\mu$ M) on TNF-$\alpha$ secretion from HMC-1, a trypsin-stimulated human leukemic mast cell line.

• Journal of Microbiology and Biotechnology
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• v.11 no.6
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• pp.1077-1086
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• 2001

The sprA and sprB gene encoding chymotrypsin-like proteases Streptomyces griseus protease A (SGPA) and Streptomyces griseus protease B (SGPB) and the sprT gene that encodes Streptomyces griseus trypsin (SGT) were cloned from Streptomyces griseus ATCC10137 and overexpressed in Streptomyces lividans TK24 as a heterologous host. The chymotrypsin activity of tole culture broth measured with the artificial chromogenic substrate , N-succinyl-ala-ala-pro-phe-p-nitroanilide, was 10, 14 and 14 units/mg in the transformants haboring the sprA, sprB and sprD genes, respectively. The growth of S. lividans reached the maximum cell mass after 4 days of culture, yet SGPA and SGPD production started in the stationary phase of cell growth and kept increasing for up to 10 days of culture in an R2YE medium. The trypsin activity of the culture broth measured with the artificial chromogenic substrate , N-${\alpha}$-benzoyl-DL- arginine-p-nitroanilide , was 16 units/mg and SGT production started in the stationary phase of cell growth and kept increasing for up to 10 days of culture in an R2YE medium. The introduction of the sprA gene into S, lividans TK24 triggered the biosynthesis of pigmented antibiotics, actinorhodin and undecylprodigiosin, and induced significant morphological changes in the colonies in Benedict, R2YE, and R1R2 media. In addition, the introduction of the sprT gene also induced morphological changes in the colony shape without affecting the antibiotic production, thereby implying that certain proteases would appear to play very important and specific roles in secondary-metabolites formation and morphological differentiation in Streptomyces.

• Bulletin of the Korean Chemical Society
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• v.32 no.2
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• pp.599-604
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• 2011

In typical proteomic analysis, trypsin digestion is one of the most time-consuming steps. Conventional proteomic sample preparation methods use an overnight trypsin digestion method. In this study, we compared high-pressure cycling technology (PCT) during enzyme digestion for proteome analysis to the conventional method. We examined the effect of PCT on enzyme activity at temperatures of 25, 37, and $50^{\circ}C$. Although a fast digestion (1 h) was used for the standard protein mixture analysis, the PCT-assisted method with urea showed better results for protein sequence coverage and the number of peptides identified compared with the conventional method. There was no significant difference between temperatures for PCT-assisted digestion; however, we selected PCT-assisted digestion with urea at $25^{\circ}C$ as an optimized method for fast enzyme digestion, based on peptide carbamylation at these conditions. The optimized method was used for stem cell proteome analysis. We identified 233, 264 and 137 proteins using the conventional method with urea at $37^{\circ}C$ for 16h, the PCT-assisted digestion with urea at $25^{\circ}C$ for 1 h, and the non-PCT-assisted digestion with urea at $25^{\circ}C$ for 1 h, respectively. A comparison of these results suggests that PCT enhanced the enzyme digestion by permitting better access to cleavage sites on the proteins.

• Journal of the Korean Society of Food Science and Nutrition
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• v.18 no.4
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• pp.410-422
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• 1989

Soybean protein isolates were acylated with succinic anhydride and partially hydrolyzed with trypsin. Chemical modification decreased protein contents of samples and, in amino acid composition, tyrosine was increased comparatively. And lysine was increased remarkably by partial proteolysis. Succinylation and trypsin treatment increased the aqueous solubility and shifted the isoelectric potint that showed high pH-dependence of protein solubility. Protein solubility was influenced by salt concentration such as $NaCl,\;CaCl_2,\;NaNO_3$ and $NaH_2PO_4$. Chemical modification increased the absorption of oil and water, emulsification properties and foam capacity, but decreased foam stability, ultraviolet absorbance and bulk density. Protein-protein Interaction between soybean protein isolates and beef protein increased the emulsifying activity, emulsifying activity index and foaming properties, but it didn't have any influence on emulsion stability.

• Journal of the Korean Society of Food Science and Nutrition
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• v.27 no.1
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• pp.132-140
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• 1998

This study aimed to verify the nutritional and curative effects of protein hydroysate and optimal ratio between protein and protein hydroysate as nitrogen source in rats with cysteamine-induced duodenal ulcer. Duodenal ulcer rat model was established by intraperitoneal injections of cysteamine. Sprague-Dawley, female rats weighing approximately 200g were intrapertionealy injected twice cysteamine(13mg/100g BW) at intervals of 3hours per day. This procedure was repeated 3 times at intervals of 3 days. Animals fed on 10% casein diet for injection periods. After last injection, 5 kinds of kiets (the ratio of casein and casein hydrolysate was 100 : 0(C100), 75 : 25(CH 25), 50 : 50(CH 50), 25 : 75(CH 75), 0 : 100(CH 100)) were given. The rate were sacrificed after feeding diet, 1, 3, 5 days. Ulcer index, hexosamine content of stomach and duodeum, gastric motility, trypsin activity, blood glutathione, plasma total protein, albumin, amino-N, urinalry urea nitrogen, creatinine, hydroxyproline and retention rate of nitrogen were analyzed for nutritional effects of diet treatments. There were no differences among diet groups in the view of the growth and diet treatments. There difference of ulcer curation by diet was appeared after 3 days. The ulcer indexes of C100 and CH 25 of 3, 5 days were significantly higher than those of CH 50, CH 75 and CH 100. This result was the same as hexosamine content of stomach, plasma protein, albumin concentration and nitrogen retention rate. The more casein hydrolysate diet had, the lower trypsin activity was. The more casein gydroysate diet had, the higher excretion of hydroxyproline was. These results show that protein hydrolysate can be applied in diet therapy for the patients with gastronitestinal ulcer. It suggests that it has curative effect of diet when nitogen sources include at least over than 50% of protein hydrolysate.

• Journal of Microbiology and Biotechnology
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• v.8 no.6
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• pp.588-594
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• 1998

Lactic acid bacteria were isolated from Kimchi and screened for bacteriocin production. Strain SE1, identified as Lactobacillus curvatus sp., showed the strongest inhibitory activity against Lactobacillus delbrueckii subsp. delbrueckii. The bacteriocin was inactivated by amyloglucosidase, trypsin, or protease K treatment. However, it maintained its activity under heat treatment at $100^{\circ}C$ for 60 min. The production of the bacteriocin had a growth-related mode and decreased around the early-stationary phase. The optimum temperature for the growth of L. curvatus SE1 was $37^{\circ}C$; however, the optimum temperature for bacteriocin production was $30^{\circ}C$. The bacteriocin activity was decreased by treatment with methanol, butanol, acetone, or chloroform, however, it was not affected by treatment with ethanol, iso-propanol, or cyclohexane. The inhibitory activity of bacteriocin was stable over a wide range of pHs (2 to 11). The bacteriocin from L. curvatus SE1 killed the indicator strain by a bactericidal mode of action. The bacteriocin from L. curvatus SE1 was partially purified by ethanol precipitation and ion exchange chromatography. SDS-polyacrylamide gel electrophoresis was used to determine the molecular weight of the bacteriocin by the bacteriocin activity test. The apparent molecular mass of the bacteriocin produced by L. curvatus SE1 was about 14 kDa.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.49 no.2
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• pp.137-145
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• 2016

This study screened the biological activity of an acidified gill extract of the Manila clam Ruditapes philippinarum including antimicrobial, hemolytic, membrane permeabilization, and DNA-binding activity, and purified the antimicrobial material. The acidified gill extract showed potent antimicrobial activity against Bacillus subtilis and Escherichia coli without significant hemolytic activity, but showed no membrane permeabilization or DNA-binding ability. An antimicrobial material was purified from the acidified gill extract using C₁₈ reversed-phase and cation-exchange high-performance liquid chromatography (HPLC). Treatment of the purified material with trypsin completely abolished all of the antibacterial activity against Bacillus subtilis, suggesting that the purified material is a proteinaceous antibiotic. The molecular weight of the purified material was 2571.9 Da, but no primary structural information was obtained due to N-terminal blocking. A future study should confirm the primary structure. Our results suggest that the Manila clam gill contains proteinaceous antibiotics that have a role in first-line defense. This information could be used to better understand the Manila clam innate immune system.

• Food Science and Biotechnology
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• v.14 no.1
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• pp.49-57
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• 2005

Enterococcus faecium MJ-14, having strong antilisterial activity, was isolated from Korean fermented food, Meju. MJ-14 showed the same phenotypic characteristics, but different sugar utilization, as reference strain, E. faecium KCCM12118. It could utilize D-xylose, amygdaline, and gluconate, whereas E. faecium KCCM12118 could not. Optimal condition for bacteriocin production by E. faecium MJ-14 was at $37^{\circ}C$ and pH 7.0. Bacteriocin activity appeared in mid exponential phase and increased rapidly up to stationary phase. Activity was significantly promoted in MRS broth containing 3.0% glucose, 1.5% lactose, 2.0% peptone, or 1.5% tryptone. Bacteriocins effectively inhibited Enterococcus faecalis and Listeria spp. of Gram-positive bacteria, and Helicobacter pylori of Gram-negative bacteria, but did not inhibit yeasts and molds. They were stable against heat (for 30 min at $100^{\circ}C$), pH (3.0-9.0), long-term storage (for 60 days at 4 or $-20^{\circ}C$), and enzymatic digestion by catalase, proteinase K, papain, lysozyme, trypsin, chymotrypsin, and lipase, etc. Bacteriocin activity was completely inhibited by protease and pepsin, and 50% by ${\alpha}$-amylase. Studies on PCR detection of enterocin structural genes revealed bacteriocins are identical to enterocins A and B.

• Proceedings of the Zoological Society Korea Conference
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• 1995.10b
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• pp.83-83
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• 1995

The three chitin synthetases of Saccharomyces cerevisiae, Chs1, Chs2, and Chs3, participate in septum and cell wall formation of vegetative cells and in wall morphogenesis of conjugating cells and spores. Because of the differences in the nature and in the time of execution of their functions, the synthetases must be specifically and individually regulated. The nature of that regulation has been investigated by measuring changes in the levels of the three synthetases and of the messages of the three corresponding gnes, CDSI, CHS2, and CAL1/CSD2/DITl0l(referred to below as CAL1), during the budding cycles. For Chs1 and Chs3, posttranslational regulation, probably by activation of latent forms, appears to be predominant. Since Chs2, like Chs1, is found in the cell in the zymogenic form, a posttranslational activation step appears to be necessary for this synthetase also. The regulation mechanism was investigated to search the relationship of CAL1, CAL2 and CALJ which is involved in Chs3 activity us ing different assay methods other than previous one. Treatment of Chs3-containing membranes with detergents drastically reduced the enzymatic activity. Activity could, however, be restored by subsequent incubation with trypsin or other pro teases in the presence of UDPGlcNAc. Experiments wi th mutants in the three genes invoIved in Chs3 activity-CAL1, CAL2, and CALJ-showed that only CAL1 and CALJ are required for the proteaseelicited (zymogenic) activity. It is concluded that Chs3 IS a zymogen and that the CAL2 product funct ions as its activator.ivator.