• Korean Journal of Microbiology
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• v.30 no.4
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• pp.239-245
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• 1992

Buci1llr.s 11c~h~n~1rnSiSi.As 3-2MI which is responsible for the special taste of traditionalKorean soy sause produced two kinds of proteinase. The activity of the proteinasc I washigher about two fold than that of proteinase 11. The optimai, reaction pH of proteinaseI and I1 wcre found to be 7-1 1.5 and 7-9. respectively. Proteinase I1 was more stable andactive than proteinase I at pH ranges around 3 to 5. The optimal te~tlperature of proteinaseI and I1 were 502. The temperature stabilitl of proteinase I1 was Inore stable thanproteinase 1 at temperature range around 30-quot;~A. ctivities of proteinase I and I1 graduallydeclined above $30^{\circ}$C and 45C. respectively. Proteinasc 1 was more active than proteinaseI1 at salt concentration range around 25-3500. The K,,, values of casein and soy proteinfor proteinase I were 6.89 mglml and 3.98 mglml. In case of proteinase 11. they were 9.00mgiml anti 11.44 111g/ml. respectively. The activity of the crudc enzyme was increased by1 rnM Pb(CH3COO). but was decreased by 5 n1M and 10 rnM of HgS04 and ZnS04. Thetwo proteinases produced amino acids and peptides from the soybean protein. The peptideswere digested into amino acids. Both protcinases were found to be the main enzymes thatproduced amino acids which make the main taste of traditional Korean soy sauce.al Korean soy sauce.

• Restorative Dentistry and Endodontics
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• v.25 no.4
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• pp.509-526
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• 2000

It is known that injuries to the dentin have a corresponding inflammatory effect on the pulp and these inflammatory effects frequently result in pulpal pathoses due to progressive degradation of pulpal connective tissue. It was supposed that the tissue degradation in different inflammatory process was controlled by proteinase activity and antiproteinase activity. Therefore, the purpose of this study was to examine the pulp and periapical pathoses in terms of the activities of proteinase and proteinase inhibitor, 37 pulpal tissues were divided by clinical diagnostic criteria into normal pulp, acute inflamed pulp, and chronic inflamed pulp, and then those groups were subdivided by histopathological findings into 5 pulpal pathoses groups, i.e. normal pulp (P1, n=8), chronic pulpitis with fibrotic change (P2, n=2), chronic pulpitis with dystrophic calcification (P3, n=11), chronic pulpitis with pulp abscess (P4, n=7), acute pulpitis with necrotic change (P5, n=4), 26 periapical tissues were also divided by ordinary histopathological findings into 3 periapical pathoses group, i.e., granuloma (A1, n=17), cyst (A2, n=2) and abscess (A3, n=7). The activities of proteinases (cathepsin G, MMP-3) and proteinase inhibitors (${\alpha}1$-AT, TIMP-1 and, SLPI) were evaluated by RT-PCR and immunohistochemical methods. The results were as follows. 1. Generally, the intensity of immunohistochemical staining of proteinases and proteinase inhibitors increased in P2 and P5 groups compared to P1 group. 2. The immunohistochemical stain of proteinases and proteinase inhibitors was intensely detected in P2 group, showing low inflammatory reaction and low tissue degradation, but it was reduced in P3 and P4 groups, showing severe tissue degradation. 3. The distribution of proteinases and proteinase inhibitors in pulpal pathoses was consistently presented by immunohistochemical staining, while the expression of proteinase and/or proteinase inhibitors mRNAs in pulpal pathoses was occasionally detected by RT-PCR methods. 4. RT-PCR of proteinase and proteinase inhibitors was usually positive in P2, showing rare tissue degradation, but it was almost negative in P3 and P4, showing severe tissue degradation. 5. We presume that the reason why the level of proteinase and proteinase inhibitors was so sparse in RT-PCR method is due to the abrupt decrease of mRNA synthesis or degradation of synthesized mRNA of proteinase and/or proteinase inhibitors depend on the inflammatory reaction and/or on the degradation of pulp tissues(P3, P4). 6. Pulpal pathoses groups showed significant lower RT-PCR detection of proteinases and proteinase inhibitors than the periapical pathoses group(p<0.05), and there is no significant difference among the periapical pathoses groups(p>0.05).

• Korean Journal of Microbiology
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• v.7 no.3
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• pp.115-124
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• 1969

The mash of Takjoo, Korean flour wine, is fermented through two brewing processes ; the primary brewing process to saccharify and the main one to produce ethyl alcohol. The activities of acid proteinase (pH3), weak acid proteinase (pH 6), and alkaline proteinase (Ph 80 on the processes are determined with time by the Folin phenol method as a strength of casein digestion. Hydrogen ion concentration, the content of total organic acids, protein, free amino acids and oligopeptides, which effect the activities of proteinase, are also measured. The results are briefly summarized as follows : 1. In general, the activities of acid proteinase and weak acid proteinase in the mesh of primary brewing process are stronger than those in main brewing process. 2. The activities of acid proteinase are remarkably stronger than those of weak acid proteinase in both processes. It reveals that they decrease slowly through the fermentation. Activities of alkaline proteinase are weaker than others. 3. As the raw materials are mixtured, the total amount of organic acids is equivalent to 0.150 mg/ml acetic acid in the mesh of primary brewing process and 0.02 mg/ml acetic acid in the main one. They increase gradually with time. 4. Hydrogen ion concnetration shows 3.9 in the mesh of main brewing process and 3.28 in the primary one. They increase to the maximum in 60-72 hrs., and decrease since 108 hrs. 5. The content of crude protein shows 66.90mg/ml in the mesh of main brewing process, while shows 64.29mg/ml in the mesh of primary one. they decrease slowly with time. it seems that a small content of crude protein, as a substrate, converts into amino acids and soluble nitrogen compounds by proteinase. 6. The content of free amino acids and oligopeptides shows 0.36 mg/ml in the mesh of primary brewing process and 0.24mg/ml in the main brewing process. It is evident that the reason they increase continuously through the fermentation is the effect of proteinase. 7. According to the results, the strong activities of proteinase in primary brewing process has been derived from the decrease of hydrogen ion concentration due to the production of organic acids.

• Journal of Nutrition and Health
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• v.6 no.4
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• pp.55-60
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• 1973

To determine the effects of condiments upon Proteinase Activity, condiments such as welsh onion, garlic, ginger, black pepper, red pepper, mi-won (glutamic acid natrium), sugar, mustard and horse-radish were ground by a homogenizer, and each of them was dosed by 0%, 1%, 5% and 10% into Pancreatin Solution of 0.2% for storage at the temperature of 15 degrees Cels. The Enzyme Solution thus obtained then was measured at a certain interval of time by the Fuld Gross Method, and the following results were obtained. 1) The condiments that kept Proteinase Action of Pancreatin checked below 75% were mustard, horse-radish, red pepper and welsh onion. The centrol power of welsh onion, in particular, became stronger as storage time became longer. 2) The condiments that kept Proteinase Action of Pancreatin checked below 50% were sugar, black pepper and ginger. 3) Mi-won and garlic showed a strong checking powor over Proteinase Action at an early stage of storage, but as time passed, their control power gradually diminished to naught. In short, it may be concluded that ail of the condiments used in this experiment demonstrated their checking power over Proteinase Action.

• YAKHAK HOEJI
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• v.5 no.1
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• pp.51-55
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• 1960

Some enzymatic properties of Cynthia roretzi V. Drasche (Korean:U-Rung-Shei) was studied by author and obtained the following results; 1. The optimum pH of the digestive gland proteinase ws 7.4-7.6 2. Activity of metallic ion on the Proteinase showed following order; 10$^{-3}$ M. M $n^{++}$>1-$^{-3}$ M. $Co^{++}$>10$^{-4}$ M. $Mg^{++}$\ulcorner10$^{-2}$ M.S $r^{++}$. Inhibition of metallic ion on the Proteinase showed following order: 10$^{-3}$ M. A $g^{+}$>10$^{-3}$ M. c $d^{++}$>10$^{-3}$ M. P $b^{++}$>10$^{-3}$ M. Z $n^{++}$ 3. The digestive gland enzyme inactivated at 70.deg. C, but no influence at 50.deg. C. 4. When the enzyme concentration increase 2 times, and 3 times, the enzymatic activity also increase, but not proportionally 5. The digestive gland Proteinase showed remarkably higher enzymatic activity than the intestinal Proteinase. 6. The digestive gland amylase brom the ascidion showed remarkably higher enzymatic activity than the heptaponcreatic amylase from shell fish (Turbo (Batillus) Cornutus Solander).).er).).).er).).

• Korean Journal of Microbiology
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• v.34 no.3
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• pp.82-86
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• 1998

An alkaline proteinase secreted from Yarrowia lipolytica 504D was purified by salting-out and column chromatography. The molecular weight of the purified enzyme was about 32,000 Da estimated by SDS-PAGE. The optimal condition for the activity of the enzyme was at pH 9.5 and $42^{\circ}C$ The enzyme was stable up to $45^{\circ}C$ and at the range of pH 4-10. Because the enzyme was inhibited by PMSF as well as EDTA, EGTA, and phenan-throlin, it is uncertain whether the enzyme is serine proteinase or metalloproteinase. However, almost all metal salts tested did not increase the enzyme activity, and Ca salt restored the activity of the enzyme inactivated by EDTA. Therefore, the purified enzyme seems to be an serine proteinase (E.C. 3.4.21.14).

• Tuberculosis and Respiratory Diseases
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• v.56 no.3
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• pp.289-296
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• 2004

Background : Mucus hypersecretion in the patients with airway diseases represents poor prognosis as well as discomfort. However, there is no known therapy for its effective control. One important component of mucus is mucin, a glycosylated protein, which endows mucus with viscosity. We studied whether a proteinase has a role in mucin secretion and if so, which. Methods : (1) Inhibition of mucin secretion Group-specific proteinase inhibitors were tested to evaluate whether a proteinase belonging to a group of proteinases plays a role in mucin secretion. Phenylmethylsulfonyl fluoride(PMSF, a serine proteinase inhibitor), E-64(a cysteine proteinase inhibitor), Pepstatin(an aspartic proteinase inhibitor) and 1, 10-Phenanthroline(a metalloproteinase inhibitor) were treated into the Calu-3 cell line for 24 hours. The enzyme linked immunoabsorbant assay(ELISA) for MUC5AC was performed to evaluate the amount of mucin secretion and to compare with a control. (2) Stimulation of mucin secretion Matrix metalloproteinase-9(MMP-9), MMP-12 and TACE(TNF-alpha converting enzyme), which are known to be related with airway diseases, were used to be treated into Calu-3 for 24 hours. ELISA for MUC5AC was performed to evaluate the amount of mucin secretion and to compare with the controls. Results : (1) PMSF($10^{-4}M$), E-64($10^{-4}M$), Pepstatin($10^{-6}M$) and 1, 10-Phenanthroline($10^{-4}M$) reduced the MUC5AC secretion by $1{\pm}4.9%$(mean${\pm}$standard deviation; P=1.0 compared with the control), $-6{\pm}3.9%$(P=0.34), $-13{\pm}9.7%$(P=0.34) and $41{\pm}8.2%$(P=0.03), respectively. (2) The amounts of MUC5AC secretion stimulated by MMP-9(250ng/ml), MMP-12(100ng/ml) and TACE(200ng/ml) were $103{\pm}6%$(P=0.39), $102{\pm}8%$(P=1.0) and $107{\pm}13%$(P=0.39), respectively, compared with the controls. Conclusion : Metalloproteinase(s) is (are) suggested to play a role in mucin secretion. It appears that metalloproteinases, other than MMP-9, MMP-12 or TACE, affect the mucin secretion in this in vitro model.

• Parasites, Hosts and Diseases
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• v.44 no.4 s.140
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• pp.321-330
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• 2006

The pathogenic mechanism of granulomatous amebic encephalitis (GAE) and amebic keratitis (AK) by Acanthamoeba has yet to be clarified. Pretense has been recognized to play an important role in the pathogenesis of GAE and AK. In the present study, we have compared specific activity and cytopathic effects (CPE) of purified 33 kDa serine proteinases from Acanthamoeba strains with different degree of virulence (A. healyi OC-3A, A. lugdunensis KA/E2, and A. castelianii Neff). Trophozoites of the 3 strains revealed different degrees of CPE on human corneal epithelial (HCE) cells. The effect was remarkably reduced by adding phenylmethylsulfonylfluoride (PMSF), a serine proteinase inhibitor. This result indicated that PMSF-susceptible proteinase is the main component causing cytopathy to HCE cells by Acanthamoeba. The purified 33 kDa serine proteinase showed strong activity toward HCE cells and extracellular matrix proteins. The purified proteinase from OC-3A, the most virulent strain, demonstrated the highest enzyme activity compared to KA/E2, an ocular isolate, and Neff, a soil isolate. Polyclonal antibodies against the purified 33 kDa serine proteinase inhibit almost completely the proteolytic activity of culture supernatant of Acanthamoeba. In line with these results, the 33 kDa serine proteinase is suggested to play an important role in pathogenesis and to be the main component of virulence factor of Acanthamoeba.

• Microbiology and Biotechnology Letters
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• v.15 no.2
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• pp.129-134
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• 1987

Aspergillus sp. (MK-24) producing a biological active substance that inhibited the venom proteinase activity was isolated from soil. The substance also inhibited the activity of trypsin and coagulation of blood, but did not inhibit papain, $\alpha$-chymotrypsin and pepsin. The substance was partially purified from culture filtrate by precipitaion with acetone, and by chromatography of DEAE-Sepadex A-50 column and Amberlite IRC-50 ion exchange. The inhibitory substance was stable in the wide pH range from 2.0 to 12.0 at 37$^{\circ}C$, but not stable at $65^{\circ}C$ in the alkaline pH. Only 12% of the activity was decreased by the heat treatment at 10$0^{\circ}C$ for two hours. The inhibition on venom proteinase (Agkistrodon bromohoffi brevicaudus) was a mixed type. The inhibitory activity depended on the preincubation time and completely depressed by cupric, zinc and cobalt ions. The inhibition on the venom proteinase was appeared strongly on casein but not on ovalbumin or hemoglobin as a substrate.

• BMB Reports
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• v.31 no.3
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• pp.303-306
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• 1998

Serine proteinase cDNA fragment from protozoan parasite Acanthamoeba culbertsoni was amplified by the reverse transcription-polymerase chain reaction (RTPCR) using degenerate oligonucleotide primers derived from conserved serine proteinase sequences. The amplified DNA fragment was subcloned and sequenced. The sequence analysis and alignment showed significant sequence similarity to other eukaryotic serine proteinases and conservation of the His, Asp, and Ser residues that form the catalytic triad. The cDNA fragment was cloned into the pGEMEX-1 expression vector and expressed in Escherichia coli. A resulting fusion protein of 56 kDa had proteolytic activity. The fusion protein reacted with sera of mice immunized with purified serine proteinase of A. culbertsoni in Western blot. Immune recognition of the fusion protein by mouse antisera suggested that the fusion protein may be valuable as a diagnostic reagent.