• Microbiology and Biotechnology Letters
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• v.24 no.3
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• pp.316-320
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• 1996

An alkaline proteinase produced by Yarrowia lipolytica TH65 was purified by 40-65% ammonium sulfate fractionation, DEAE-cellulose chromatography, and gel filtration with Sephadex G-100 and Sephadex G-75. The purified enzyme was shown as a single band on SDS-PAGE, and its molecular weight 31,500. Optimum temperature and pH were 40$\circ$C and 8.5-9.0, respectively, and the enzyme was stable below 40$\circ$C and in the pH range of 6-8. The enzyme was strongly inhibited by divalent ions, completely by PMSF, and partially by EDTA, EGTA, and phenanthroline. But the inhibitory effect in the presence of EDTA, EGTA and phenanthroline could be reversed by addition of Ca$^{2+}$. Thus, these results indicated that the purified enzyme was an alkaline serine proteinase (E.C. 3.4.21.14).

• Applied Biological Chemistry
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• v.33 no.4
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• pp.287-292
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• 1990

Eight(I through VII) of Bowman-Birk proteinase inhibitor have been isolated from soybean with DEAE-Sephadex A-50. Inhibitor VII was a typical BBTI, showing high cysteine content(17%/mole) ud low trypsin to chymotrypsin inhibiting activity(TIA/CIA= 1.0) with the independent reactive site to each enzyme. Dissociation constant of trypsin-BBTI and chymotrypsin-BBTl complexes were $9.17{\times}10^{-9}M$ and $5.14{\times}10^{-8}M$, respectively. Inhibitor Vll was extremely heat stable. Six hours heat treatment at $100^{\circ}C$ caused only 50% decrease in it's original inhibiting activity. Except inhibitor III,6 other isoinhibitors differed from a typical BBTI in TIA/CIA, values, ranging from 3 to 29.

• BMB Reports
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• v.41 no.11
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• pp.814-819
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• 2008

Interleukin-32 (IL-32) induces a variety of proinflammatory cytokines and chemokines. The IL-32 transcript was reported originally in activated T cells; subsequently, it was demonstrated to be abundantly expressed in epithelial and endothelial cells upon stimulation with inflammatory cytokines. IL-32 is regulated robustly by other major proinflammatory cytokines, thereby suggesting that IL-32 is crucial to inflammation and immune responses. Recently, an IL-32$\alpha$-affinity column was employed in order to isolate an IL-32 binding protein, neutrophil proteinase 3 (PR3). Proteinase 3 processes a variety of inflammatory cytokines, including TNF$\alpha$, IL-$1{\beta}$, IL-8, and IL-32, thereby enhancing their biological activities. In the current study, we designed four PR3-cleaved IL-32 separate domains, identified by potential PR3 cleavage sites in the IL-32$\alpha$ and $\gamma$ polypeptides. The separate domains of the IL-32 isoforms $\alpha$ and $\gamma$ were more active than the intrinsic $\alpha$ and $\gamma$ isoforms. Interestingly, the N-terminal IL-32 isoform $\gamma$ separate domain evidenced the highest levels of biological activity among the IL-32 separate domains.

• Korean Journal of Food Science and Technology
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• v.7 no.4
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• pp.229-237
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• 1975

An attempt was made to utilize the enzyme produced by Asp. oryzae as meat tenderizer. The production, purification, and various properties of proteinase produced by Asp. oryzae were investigated. Results obtained are as follow; 1. A strain which had the highest proteolytic activity was selected among 9 Aspergillus species. 2. Culture medium consisted of wheat bran 10g, 2% glucose, 0.03% urea and 0.1% $MgSO_4$ (pH 6.5). Mold was incubated at $30^{\circ}C$ for 3 days. 3. Enzyme extract from culture medium were fractionated with ammonium sulfate and purified by Sephadex G-75 column chromatography. 4. When pH of reaction mixture was controlled, maximal activity of proteinase by Asp. oryzae was obtained at pH 3, pH 6.6, $8.4{\sim}8.5$ and pH 10.0 to 10.5. Those results were interpreted to show that enzyme consists of acid proteinase, neutral proteinase and alkaline proteinase. Enzyme was stable at pH 6 to 10. 5. Opt. temperature for proteinase activity was $50^{\circ}C$, but enzyme was stable up to $40^{\circ}C$. 6. The proteinase was inhibited by $Ag^+$. It was also inhibited by EDTA. 7. When myofibrillar proteins were treated by proteinase from Asp. oryzae, ATPase activities of myofibrillar proteins changed remarkably. Accordingly, it was concluded that proteinase produced by Asp. oryzae were able to be used as meat tenderizer.

• The Korean Journal of Mycology
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• v.14 no.2
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• pp.101-107
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• 1986

The properties of carboxyl proteinase which was contained in Poria cocos (Schw.) Wolf were investigated by means of the purification with 0.65 ammonium sulfate saturation, DEAE cellulose and Sephadex G-75 gel filtration. This enzyme was found to hydrolyze only peptide bond between glutamyl-L-tyrosine of carbobenzoxy-L-glutamyl-L-tyrosine among the synthetic substrates of carbobenzoxy-L-glutamyl-L-tyrosine, hippuryl- L-phenylalanine and hippuryl-L-arginine. This enzyme was inhibited by $Zn^{+2},\;Fe^{+2},\;Ca^{+2},\;CN^{-1},\;P_2O_7^{-4}$ ions, but stimulated by $Hg^{+2}$ ion. Also, this enzyme was inhibited by organic compounds such as L-lysine, L-phenylalanine, hippuryl-L-phenylalanine, diazoacetyl-DL-norleucine methyl ester (DAN) and 1,2-epoxy-3-(P-nitrophenoxy)propane(EPNP). In particular, the activity was inhibited by L-lysine till 20 minutes of preincubation time rapidly, and by DAN in the presence of $Cu^{+2}$ ion more rapidly after 30 minutes than DAN in the absence of $Cu^{+2}$ ion. L-Lysine was found to be a competitive inhibitor and its $K_i$ value was determined to be 0.12 mmole by Dixon plot.

• Biomedical Science Letters
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• v.8 no.3
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• pp.161-165
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• 2002

Apoptosis can be difficult to detect in routine histological sections. Since extensive DNA fragmentation is an important characteristic of this process, visualization of DNA breaks could greatly facilitate the identification of apoptotic cells. Several techniques for the qualitative and quantitative detection of this process have been established; recently, an in situ nick end-labelling technique based on the detection of DNA fragmentation, which is a molecular characteristic of apoptotic cell death, was described. Applying this method to paraffin sections of rat tissues, sensitivity was observed to be inconsistently low with regard to the expected number of apoptotic cells. I describe a new modified method for formalin-fixed, paraffin-embedded tissue sections, pretense pretreatment to permeate the tissue sections that involves an TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling) is acknowledged as a method of choice in the rapid identification and quantification of the apoptotic cell fraction in paraffin tissue preparations. TUNEL was performed without apoptosis and with apopotosis samples to each of the three concentrations of proteinase K (10, 25, 40 mg/ml) pretreatments. In this study, I show that chemical pretreatments of the tissue sections in proteinase K (25 mg/ml for 15 min at room temperature) considerably enhances the sensitivity of this nick end labelling technique.

• Journal of Chest Surgery
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• v.27 no.5
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• pp.364-373
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• 1994

Alpha 1-Proteinase inhibitor[PI] was known as a major protective enzyme against to excessive hydrolytic and proteolytic reaction. So, it was suggested that Alpha 1-PI may implicated in growth of bronchogenic cancer. This study was undertaken to investigate the role of Alpha 1-PI in local invasion of bronchogenic cancer. Three groups of patients were studied; Preliminary research group of 15 bronchogenic cancer patients, Main research group of 13 bronchogenic cancer patients and Normal control group of 10 nephrectomy donor. Serum Alpha 1-PI level was observed in each group of patients during pre-and postoperative days. Pre-operative serum Alpha 1-PI level in preliminary research group [329.2$\pm$14.21mg/dl]and main research group[406.2$\pm$39.30mg/dl] were higher than in normal control group[236.2$\pm$19.55mg/dl] significantly[p<0.005]. Serial Alpha 1-PI level in each group during pre-and postoperative days shows peaked at 3rd. postoperative day in preliminary and main research group, thereafter decreased gradually. Immunohistochemical study for Alpha 1-antitrypsin[A1AT] was carried out by ABC[avidin-biotin peroxidase complex] method using Alpha-1 antitrypsin DAKOR to tumor tissues of 13 lung cancer patients in main research group. 6 cases[46.2%, squamous cell ca.;5, adenocarcinoma;1] of above 13 cases show positive immunoreactivity for A1AT. In conclusion, alpha 1-PI and elastase are disclosed that have defined actions for lung cancer growing or spreading.

• Development and Reproduction
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• v.10 no.4
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• pp.263-269
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• 2006

To evaluate the potentiality of industrial use of cysteine proteinase inhibitor (cystatin) of tilapia egg and serum stability of the tilapia cystatin on low temperature storage and heat treatment was studied. When the eggs were stored at $4^{\circ}C$ for 3 days the cystatin activity was not changed much, while the supernatant of egg homogenate lost its cystatin activity significantly, remaining only about 65% of initial activity. When the eggs and serum were subjected to repeated freeze at $-20^{\circ}C$ and thaw at room temperature once a day, the egg cystatin was decreased after 5 cycles of freeze and thaw. However the serum cystatin was not changed by the 5 times repetition of freeze and thaw. More than 80% of egg cystatin activity was remained when the egg was heated at $35^{\circ}C$ for 30 min, but less than 10% was remained when heated at $50^{\circ}C$. On the other hand, the serum cystatin was very resistant to heat, remaining about 74% after heating at as high as $80^{\circ}C$ for 30 min. In summary, the egg cystatin was more stable when stored as intact form of egg rather than as supernatant of homogenate when stored at refrigeration. Egg cystatin was relatively stable against repeated freeze-thaw, and serum was found to be more stable in cysteine proteinase inhibitory activity than egg. Egg cystatin was not very resistant to heat treatment, while serum cystatin was quite resistant to high temperature heat treatment. These results suggest that tilapia egg and serum, especially the serum, would be a useful source for cysteine proteinase inhibitor in surimi production.

• Journal of Life Science
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• v.8 no.3
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• pp.333-338
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• 1998

Productivity of alkaline proteinase from Yarrowia lipolytica 504D was investigated. For the production fo the enzyme, hemoglobin was the best nitogen source, however, casein and skim milk were also good. All carbon sources inhibited strongly the producitivity of the enzyme. Yeast extract increased the productivity of the enzyme to 220%, but almost mineral salts except monovalant ions decreased it. Based on these results, optimal medium was composed of 1.2% casein, 0.2% glucose, 0.16% yeast extract, and 0.1% ammonium sulfate. the best condition for the production of the enzyme was observed at pH 9 and $20^{\circ}C$ for 42 hours.

• Journal of Nutrition and Health
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• v.6 no.1
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• pp.39-46
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• 1973

This study was projected to get basic data which can provide a basis for future direction in nutritional education, and also to find the way how to improve the nutritional supply by evaluating the current nutritional intake of average high school students through the survey study of their daily packed lunch. Five hundred twenty seven students from two boys high school and two girls high school including one general and one vocational school respectively were chosen as random sampling technique. Four hundred forty nine among the 527 students had brought lunch. The contents of lunch box were weighed and converted into nutritional values according to the food composition table and compared with recommended dietary allowances. The results compared and classified by sex, School and housewives' educational level were as follows: 1. The nutritional supply in the lunch box was 671 Cal of energy and 22.3 gm of protein for male students which were respectively 55.9% and 74.2% of the dietary recommendations. On the other side female student's lunch boxes were found to contain 495 Cal of energy and 21.3gm of protein which are respectively 61.8% and 80% of the dietary prescriptions. Excluding niacin, all vitamins and minerals were found to be short. 2. Calorie intake in the vocational high school was found to be higher than in the general high school but lower in protein intake especially significant difference (P<0.01) in animal protein. 3. From the nutritional point of view the educational backgrouud of the housewives was not found to have any influence in the way of preparing the lunch boxes. 4. Nutrients of lunch box were heavily inclined to grain rather than to side dishes.