• Journal of Korean Society of Environmental Engineers
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• v.36 no.4
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• pp.265-270
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• 2014

Protease producing microorganisms were isolated from many kinds of food waste and fermented foods, which contains high amount and variable kinds of degraded substances. Several microorganisms were identified by 16S rRNA full sequencing analysis methods. The activity of protease was analyzed and identified in variable conditions for the application. For industrial use for biowaste treatment some proteases were isolated, identified and selected from microbial cells. And the tests were carried for the further use. The protein degrading activity at low temperature is useful for the treatment of organic waste, which contains much proteins. By the protein degradation process the organic waste can be utilized in variable fields, for example from feedstuff supplement to fertilizer for agriculture. Bacterial cells with protease activity at low temperature were isolated and identified. The optimal conditions for microbial cultivation and protease production were studied.

• KSBB Journal
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• v.14 no.2
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• pp.131-135
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• 1999

Effects of protease, $CaCl_2$ EGTA, polybrene, medium pH, and formaldehyde on the infection and inactivation of human rotavirus Wa were investigated using T-flask culture of monkey kidney MA-104 cells. Rotavirus titer was improved by the addition of trypsin or clostripain. Rotavirus titer was increaeed 8 and 10 time sin the infection medium supplemented with 300 $\mu\textrm{g}$/mL of $CaCl_2$ and in the medium adjusted its pH to 8, respectively. However, addition of EGTA or polybrene to the medium decreased rotavirus titer. Rotavirus titer was reduced to 53-95% of the initial value at 1 hr after formaldehyde treatment. Furthermore, rotavirus was inactivated more than 98% at 12 hrs after formaldehyde treatment.

• Preventive Nutrition and Food Science
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• v.2 no.2
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• pp.109-120
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• 1997

In order to elucidate roles of lactic acid bacteria(LAB) for the antibiosis occurring in th fermenting environment of Kimchi, 2.052 strains of LAB were isolated from Kimchi. Fifty tow strains which showed antagonistic effect against 4 indicator strains were finally selected and investigated. Based upon responses to protease treatment, antibiosis of the 52 strains of LAB were classified into 3 types. Type A antibiosis resulted from action of antibiotic-like substances which were not affected by protease treatment and which had broad action spectra against even natural inhabitants of Kimchi. Type B antibiosis was due to bacteriocin-like substances which were very sensitive to treatment of protease and more effective against foreign bacteria than original inhabitant microflora. Type C antibiosis was owing to proteinaceous compounds which were activated or induced by the presence of protease and then exerted antibacterial activities. Therefore, lactic acid bacteria appeared to contribute to antibiosis of Kimchi by the concerted action of these three different types of antibacterial compounds. As one of model system for type B bacteriocin, the antagonistic compound produced by LAB31-9 as well as th producer strain itself was further charaacterized. Strain LAB31-9 was identified as L. casei. Bacteriocin produced by LAB31-9 was proteinaceous and stable over wide range of pH and to various solvents, but very labile to heat treatment. Its mode of action was bactericidal. Based upon these data, bacteriocin produced by LAB31-9 was named as 'caseicin K319'. Genetic determinant for the bacteriocin production of LAB31-9 was located in the chromosome.

• Journal of the Korean Society of Clothing and Textiles
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• v.22 no.7
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• pp.843-850
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• 1998

The purpose of this study is the investigation of chemical properties of wool treated with oxidants and protease at low temperature. The chemical degradation of the fibers were investigated by measuring $\alpha$-amimo acid contents and FT-IR analysis. In addition, urea-hydrogensulfite solubility was measured to compare to the oxidation and protease treated wool. The results were as follows. 1) By the oxidation of wool, cystine is oxidised to cysteic acid by way of the intermediate oxides, cystine-S-monooxide and cystine-S-dioxide, in the case hydrolysis catalysed by the protease catalyse. Also, $\alpha$-amimo acid contents is increased, and urea-hydrogensulfite solubility was lower than that of untreated wool. This chemical degradation of wool was occurred due to oxidate hydrolysis in the order of permonosulfate>dichloroisocyanuric acid$\geq$chlorine. 2) The chemical degradation of wool was accelerated by the protease treatment of oxidized wool. Oxidation of wool is considered to make the fiber more susceptibled to enzymatic attact by opening disulphide bond within wool. Enzymatic attact was effectively directed to the wool oxidised by permonosulfate.

• Korean Journal of Microbiology
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• v.38 no.4
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• pp.235-240
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• 2002

Protease producing bacterium, PB16 was isolated from food processing wastewater sludge and paddy field soil samples and selected by the clear zone and enzyme activity test. The isolate was gram negative, rod type and its protease productivity was 6.49 U/ml. As a result of API20NE kit test and 16S rDNA sequencying, the isolated PB16 was identified as Aeromonas hydrophila (99%). The growth rate ($h^{-1}$) was 0.21 in synthetic waste water only and 0.26 in synthetic waste water containing vitamin and mineral using a bioscreen C. Synthetic wastewater removal rate was 59 and 87%, respectively after 1 and 3 day reaction (intial CODcr was 2,472 mg/l).

• Journal of Microbiology and Biotechnology
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• v.28 no.2
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• pp.275-283
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• 2018

Ischemic stroke can result from blockage of blood vessels, forming fibrin clots in the body and causing irreparable brain damage. Remedial thrombolytic agents or anticoagulants have been studied; however, because the FDA-approved tissue plasminogen activator has low efficacy and side effects, it is necessary to develop safer and more effective treatment candidates. This study aimed at assessing the fibrinolytic and anticoagulation features of a novel serine protease extracted and purified from Diopatra sugokai, a polychaeta that inhabits tidal flats. The purified serine protease was obtained through ammonium sulfate precipitation, affinity chromatography, and ion-exchange chromatography. Its molecular size was identified via SDS-PAGE. To characterize its enzymatic activities, the protease activity at various pH and temperatures, and in the presence of various inhibitors, was measured via azocasein assay. Its fibrinolytic activity and anticoagulant effect were assessed by fibrin zymography, fibrin plate assay, and fibrinogenolytic activity assays. The novel 38 kDa serine protease had strong indirect thrombolytic activity rather than direct activity over broad pH (4-10) and temperature ($37^{\circ}C-70^{\circ}C$) ranges. In addition, the novel serine protease exhibited anticoagulant activity by degrading the ${\alpha}$-, ${\beta}$-, and ${\gamma}$-chains of fibrinogen. In addition, it did not produce cytotoxicity in endothelial cells. Therefore, this newly isolated serine protease is worthy of further investigation as a novel alkaline serine protease for thrombolytic therapy against brain ischemia.

• Korean Journal of Food Science and Technology
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• v.27 no.3
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• pp.370-374
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• 1995

By exposing the complex enzyme solution to alkaline condition, it was possible to remove the protease activity selectively without inactivation of soybean cell wall degrading activity of the crude enzyme complex produced by Aspergillus niger CF-34. Optimum reaction conditions were as follow. pH was $9.0{\pm}0.1$, temperature was $20^{\circ}C$ and reaction time was 30 min with gentle stirring. Over 90% of protease activity could be eliminated while the activities of pectinase, polygalacturonase, xylanase, carboxymethyl cellulase and soybean cell wall degrading enzyme were maintained to $80{\sim}100%$. Through alkali treatment, it was discovered that the quality and organoleptic properties of soy protein produced by this enzymes were improved because the hydrolysis of protein and formation of bitter peptide were decreased.

• Journal of the Korean Society of Food Science and Nutrition
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• v.23 no.1
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• pp.78-89
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• 1994

To evaluate of quality of Yakju brewed with different types of fermenters(Nuruk) and brewed by a method described in Sallymkyungjae, blucoamylase and protease and protease activities of Nuruk were measured, and proximate composition , coloring degree, acetaldehyde , alcohol , , fusel oils , amino acid content , mineral content and sugar composition of Yakju samples were also determined. It was foung that the lower the pH, the lower the glucoamylase acitvity in Nuruk samples A to E . In A, B, D and H, protease activity washigher at ph 5.5 than at pH 5.0 . Yakju sampels 1 to 5 during fermentation , total acidity and amino acid content were high at the first step and were getting lower gradually at the second step. In Yakju samples 6 to 11 , ethanol content was high in 6b, 7b, 8a, 8b and 11b. At the second step, residual sugar content was getting higher gradually in 7 a, 11 a and 11b. The coloring degree of Yakju wasinfluenced by not only color of Nuruk but also Fe content in Yakju . Of the Yakju examined , only 9a and 9b contained acetaldehyde in trace amounts. Thanol content was the highest in 8a and 8b. Fusel oil content was high in 8a and 8b. In samples 6 to 9 , aspartic acid content was higher in treatment a than treatment a than treatment b, but tyrosine , histidine and proline contents were higher in treatment a than treatment b. The levels of frucose, melibiose, sorbitol, and arabinose in Yakju brewed from unsteamed rice were higher than in Yakju brewed from steamed rice. K content was the lowest in 9a and 9b. Na content was higher in treatment a than in treatment a than in treatment b. In only 6a, Ca/P ratio was more than one.

• Food Science and Preservation
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• v.13 no.5
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• pp.563-568
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• 2006

The properties of mung bean were investigated depending on hydrolysis condition. The results showed that enzyme treatments (${\alpha}$-amylase and protease each at 0.1% (w/w)) by varying hydrolysis temperature showed better properties than non-pretense treatment (control group). The treatment with 0.08% ${\alpha}$-amylase was best for optimum hydrolysis of mung bean starch The treatment using a mixture of 0.08% (w/w) ${\alpha}$-amylase and 0.12% (w/w) protease was best for optimum hydrolysis of meg bean protein. The effects of Hydrolysis time of mung bean showed that the optimum time was 60 and 90 min and there fore the optimum time was set at 60 min. These result showed that the best hydrolysis conditions of mung bean were the treatment at $60^{\circ}C$ for 60 min using a mixture of 0.08%(w/w) ${\alpha}$-amylase and 0.12% (w/w) pretense, with the sugar level shown at $5.8^{\circ}Brix$, reducing sugar at 2,022.13 mg% and crude protein at 7,666.17 mg%.

• Journal of Life Science
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• v.11 no.1
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• pp.42-46
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• 2001

Subtilisin YaB, produced by alkalophilic Bacillus strain YaB, is an extracellular alkaline serine protease having 55% homology to subtilisin BPN'. It is synthesized as a 378-amino acid preproenzyme and secreted into the culture medium as a 265-amino acid mature protease. To examine the role of pro-sequence for the secretion of subtilisin YaB, we have studied the expression, in Bacillus subtilis, of a mutant preprosubtilisin YaB in which active site Ser214 is substituted with Cys. The use of a six protease-deficient strain, WB600, was required for its efficient production. The prosubtilisin YaB, thus produced, was indeed secreted into the culture medium and was processed to its mature form upon treatment with exogenously added active subtilisin YaB. From these results, we have concluded that the processing of pro-sequence is not essential for the secretion of the enzyme.