• Journal of Periodontal and Implant Science
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• v.23 no.3
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• pp.442-453
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• 1993

In order to determine the relationship between probing pocket depth and trypsin-like activity in subgingival plaque, probing pocket depth and loss of attachment were measured by Michigan-O probe on mandibular incisors of 30 patients with adult periodontitis. And the trypsin-like activity of Treponema denticola, Porphyromonas gingivalis, and Bacteroides forsythus was evaluated by the hydrolysis of N-Benzoyl-DL-Arginine-2-Naphthyla-mide (BANA) using PerioScan reagent cards(Oral-B Laboratories, Redwood City, CA). The obtained data were statistically analyzed by Microstat program. The results were as follows. 1. The number of teeth showing negative trypsin-like activity was more in shallow periodontal pocket groups, but the number of teeth showing positive trypsin-like activity was more in deep periodontal pocket groups. 2. There was a significant positive correlation between probing pocket depth and trypsin-like activity in subgingival plaque(y=0.413X - 0.955, r = 0.7024, p<0.001). 3. There was no consistent relationship between loss of attachment and trypsin-like activity in subgingival plaque(p>0.01).

• Natural Product Sciences
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• v.10 no.5
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• pp.211-216
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• 2004

Citrus unshiu (Rutaceae) has long been known as an anti-inflammatory and anti-allergic agent. In the present study, the inhibitory effect of CUWE (Citus unshiu water extract) on the production of $TNF-{\alpha}$ and tryptase was examined. In addition, a possible mechanism for the inhibition of trypsin-stimulated human leukemic mast cell-1 (HMC- 1 ) activation was determined. To do so, $TNF-{\alpha}$ production from the HMC-1 cells that were stimulated by trypsin (100 nM) in the presence or absence of CUWE $(10,\;100,\;and\;100\;{\mu}g/ml)$ was measured by enzyme-linked immunosorbent assay (ELISA) and reverse transcription-PCR. The tryptase production was evaluated by reverse transcription-PCR. Extracellular signal-regulated kinase (ERK) activation was analyzed by Western blot. Trypsin activity was measured by using Bz-DL-Arg-p-nitroanilide (BAPNA) as substrate. Results showed that the CUWE inhibited production of both $TNF-{\alpha}$ and tryptase from the trypsin-stimulated HMC-1 in a dose-dependent manner. The CUWE a1so inhibited the ERK phosphorylation and trysin activity. These results indicate that the CUWE had an inhibitory effect on $TNF-{\alpha}$ and the tryptase productions by blocking the ERK phosphorylation and trypsin activity.

• Journal of Aquaculture
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• v.18 no.4
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• pp.245-251
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• 2005

This study was investigated the condition of their maximum activity to assay the enzymes of rotifer, Brachionus rotundiformis의 $\alpha$-amylase, total alkaline Protease, trypsin and TG-lipase activities of rotifer were higher and more sensitive in phosphate-NaOH buffer than Tris-HCl buffer. $\alpha$-amylase, trypsin and TG-lipase activities were appeared the maximum at pH 8.0, and total alkaline protease activity showed the maximum activity at pH 7.0. $\alpha$-amylase activity showed the highest activity at $40^{\circ}C$, and total alkaline protease and trypsin activities were assayed the highest at $55{\~}60^{\circ}C$. However, TG-lipase activity was appeared the highest at $25{\~}30^{\circ}C$. The optimum substrate concentration of enzyme activity of a-amylase, total alkaline protease, rypsin and TG-lipase were $3.5\%$ starch, $\0.6%$ azo-casein, $87.5{\mu}M$ BApNA and 81.2 mM olive oil, respectively. The optimum reaction time of enzyme activity of $\alpha$-amylase, total alkaline protease, trypsin and TG-lipase were increased up to 40, 60, 30 and 25 min., respectively. The data obtained in this study could be used for the digestive enzyme research of rotifer, B. rotundiformis.

• YAKHAK HOEJI
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• v.49 no.5
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• pp.410-415
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• 2005

To investigate biological activity of noni extracts treated with HCl and trypsin, we measured the antioxidant activity through vitro assay and cellular system. Both water and lipid soluble fraction of noni extracts dose-dependently scav­enged DPPH radical. Superoxide scavenging activity of lipid soluble fraction after treating HCl and trypsin was significantly more potent than those of other fractions in NBT/xanthine oxidase assay, which suggests that antioxidant activity of noni extracts was increased by the treatment with HCl and trypsin. In antioxidant assay using RBL 2H3 cells, water soluble frac­tion of noni extracts had little effect on silica-induced reactive oxygen species generation, whereas lipid soluble fraction inhibited in a dose dependent manner. In non-treated noni extracts, effect of water soluble fraction on silica/$CuSO_4$-induced lipid peroxidation was more potent than that of lipid soluble fraction. However, the effects of noni extracts were reversed in noni extracts treated with HCl and trypsin. These data suggest that water soluble substances may be converted into lipid soluble substances by the treatment with HCl and trypsin. From the above results, it is suggested that lipid soluble fraction of noni extracts contain antioxidant used in vitro assay and RBL 2H3 cellular system. Such an effect of noni extracts may be increased by the treatment with HCl and trypsin.

• Food Science and Biotechnology
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• v.18 no.5
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• pp.1173-1179
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• 2009

Trypsin inhibitors and lectins are defense proteins produced by many organisms. From Chinese 'Large Black Soybeans', a 60 kDa lectin and a 20 Da trypsin inhibitor (TI) were isolated using chromatography on Q-Sepharose, Mono Q, and Superdex 75. The TI inhibited trypsin and chymotrypsin with an $IC_{50}$ of 5.7 and $5{\mu}M$, respectively. Trypsin inhibitory activity of the TI was stable from pH 3 to 13 and from 0 to $65^{\circ}C$. Hemagglutinating activity of the lectin was stable from pH 2 to 13 and from 0 to $65^{\circ}C$. The TI was inhibited by dithiothreitol, signifying the importance of disulfide bond. The TI and the lectin inhibited HIV-1 reverse transcriptase ($IC_{50}$=44 and $26{\mu}M$), and proliferation of breast cancer cells ($IC_{50}$=42 and $13.5{\mu}M$) and hepatoma cells ($IC_{50}$=96 and $175{\mu}M$). The hemagglutinating activity of the lectin was inhibited most potently by L-arabinose. Neither the lectin nor the TI displayed antifungal activity.

• Journal of Ginseng Research
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• v.28 no.3
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• pp.127-131
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• 2004

This study was carried out to investigate the effect of red ginseng water extract (RGWE) on trypsin activity. After extraction of fat soluble and saponin component from red ginseng powder by methyl alcohol, the residue was extracted with distilled water, and manufactured to water extract. The extract was dialyzed with different molecular cut off membrane. Trypsin activity demonstrated the highest level at the RGWE concentration of 9${\times}$10$\^$-2/% in reaction mixture, and also increased to 15% at 2.9${\times}$10$\^$-3/%. Km value was decreased and Vmax was increased in the present of red ginseng water extract. Red ginseng water extract was partially purified by dialysis, Bio-Gel P-I0 and DEAE-cellulose column chromatography. The active fraction demonstrated positive reaction to ninhydrin, DNS and folin reaction.

• International Journal of Industrial Entomology and Biomaterials
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• v.8 no.2
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• pp.195-200
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• 2004

Trypsin can be immobilized on silk fibroin fiber (SFF) by introducing several spacer arms, such as ethylene diamine (ED), bovine serum albumin (BSA) and silk sericin (SS). Direct immobilization on silk fiber (SFFGA) has low activity because of the steric hindrance between the trypsin and substrate. The introduction of spacer arms onto SFF-GA can enhance the activity of trypsin by reducing the steric hindrance. When ED is used as a spacer arm, the activity of trypsin has increased but its stability decreased due to the increased hydrophobicity of SFF. BSA and SS, as a spacer arm, have better results in both activity and stability. SFF-BSA shows some decrease in the specific activity due to improper immobilizatin. SFF-SS maintained 90% of its initial activity even after 12 hrs incubation at $50^{\circ}C$. In the case of repeated hydrolysis of silk sericin with immobilized trypsin, SFF-GA and SFF-ED lost 50% of their initial activity right after first run, whereas SFF-BSA and SFF-SS maintained 80% of their initial activities even after 5 runs. Higher operational stability is due to increased hydrophilicity of SFF by introducing hydrophilic spacer arms such as BSA and SS. The high content of serine in SS increases the hydrophilicity of SFF resulting the best results among other spacer arms.

• YAKHAK HOEJI
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• v.52 no.1
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• pp.79-84
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• 2008

Alismatis Rhizoma has been used as diuretics and antiphlogistics in the Chinese oriental medicine. A trypsin inhibitor was isolated from Alismatis Rhizoma using DEAE ion exchange column, trypsin affinity column, and FPLC chromatography, and its activity and characteristics were studied. The purifed Alismatis Rhizoma trypsin inhibitor (ARTI) was estimated to be about 22 kDa. The sequence determination on N-terminal amino acid residues and 84 amino acid residues has been completed, yet no homology has been found with trypsin inhibitors reported at NCBI. ARTI did not show inhibitory activities on chymotrypsin and elastase, however it exhibited a significant inhibitory activity on bovine trypsin, and formed a complex with rat liver 10-formyltetrahydrofolate dehydrogenase.

• Fisheries and Aquatic Sciences
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• v.1 no.2
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• pp.209-215
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• 1998

Trypsin-like enzyme was purified from shrimp hepatopancreas through Q-Sepharose ionic exchange, benzamidine Sepharose-6B affinity, and Superdex 75 gel chromatography. Purity of trypsin-like enzyme was increased 69-fold with $44\%$ yield. The enzyme consisted of a single polypeptide chain with a molecular weight (M.W.) of 32 kDa judged by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme was completely inactivated by serine enzyme inhibitors such as soybean trypsin inhibitor (SBTI), tosyl-L­lysine chloromethyl ketone (TLCK), and leupeptin. However, the enzyme was not affected by tosyl-L-phenylalanine chloromethyl ketone (TPCK) which is a chymotrypsin specific inhibitor. The enzyme had no activity against benzoyl-tyrosine ethyl ester (BTEE) which is a chymotrypsin specific substrate. The enzyme showed high activity on the carboxyl terminal of Phe, Tyr. Glu, Arg, and Asp. However. no activity was detected against the carboxyl terminal of Pro, Trp, Cys, Gly, Val, and Ala.

• Natural Product Sciences
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• v.13 no.2
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• pp.169-173
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• 2007

Luteolin is a major flavonoid of Lonicera japonica and has anti-inflammatory effect. The activation of proteinase-activated receptor (PAR)-2 and -4 by trypsin appears to play a role in inflammation, In the present study, we examined the inhibitory effects of luteolin on activation of trypsin-induced human leukemic mast cells (HMC-1). HMC-1 cells were stimulated with trypsin, PAR-2 and PAR-4 agonist, in the presence or absence of luteolin. The level of TNF-${\alpha}$ secretion was measured by enzyme-linked immunosorbent assay (ELISA). The expression of tryptase and phosphorylated-extracellular signal-regulated kinase (ERK) were assessed by Westem blot analysis. Moreover, trypsin activity was measured by the substrate Bz-DL-Arg-p-nitroanilide (BAPNA). TNF-${\alpha}$ secretion and Tryptase expression in trypsin-stimulated HMC-1 cells were markedly inhibited by pretreatment of luteolin. Furthermore, the pretreatment of luteolin resulted in the reduction of ERK phosphorylation and trypsin activity. These results suggest that luteolin might has the inhibitory effects on the PAR-2 and -4-dependent inflammation.