• Journal of Acupuncture Research
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• v.19 no.2
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• pp.51-64
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• 2002

We have compared(using the same series of experimental tissue samples) the levels of proteolytic enzyme activities and free radical-induced protein damage in synovial fluid from RA and CPH cases. Many protease types showed significantly increased (typically by a factor of approximately 2-3-fold) activity in RA, compared to normal rats. However, CPH significantly reduced the cytoplasmic enzyme activities of arginyl aminopeptidase, leucyl aminopeptidase, pyroglutamyl aminopeptidase, tripeptidyl aminopeptidase, and proline endopeptidase to almost about 1/10 each. For the Iysosomal proteases, synovial fluid samples from RA rats, CPH significantly reduced the enzyme activities of cathepsin B, dipeptidyl aminopeptidase I and dipeptidyl aminopeptidase II. In extracellular matrix degrading(collagenase, tissue elastase) and leukocyte as sociated proteases (leukocyte elastase, cathepsin G), CPH decreased these enzyme activities of collagenase, tissue elastase and leukocyte associated elastase in RA. In cytoplasmic and lysosomal protease activities in plasma from RA. CPH and normal plasma samples were not significantly different, suggesting that altered activity of plasma proteases (particularly those enzymes putatively involved in the immune response) is not a contributory factor in the pathogenesis of RA. In addition, the level of free radical induced damage to synovial fluid proteins was approximately twice that in RA, compared with CPH. CPH significantly decreased the level of ROS induced oxidative damage to synovial fluid proteins (quantified as protein carbonyl derivative). Therefore we conclude that both proteolytic enzymes and free radicals are likely to be of equal potential importance as damaging agents in the pathogenesis of inflammatory joint disease, and that the design of novel therapeutic strategies for patients with the latter disorder should include both protease inhibitory and free radical scavenging elements. In addition, the protease inhibitory element should be designed to inhibit the action of a broad range of protease mechanistic types (i.e. cysteine-, metallo- and serine- proteinases and peptidases). However, increased protein damage induced by ROS could not be rationalised in terms of compromised antioxidant total capacity, since the latter was not significantly altered in RA synovial fluid or plasma compared with CPH.

• Food Science of Animal Resources
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• v.33 no.6
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• pp.772-780
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• 2013

The aim of this study was to investigate the effect of season and location on activities of enzyme produced by psychrotrophic bacteria isolated from raw milk located in Kyunggi region of South Korea. Agar diffusion and colorimetric methods were used for the lipase and protease activities of psychrotrophic bacteria. Intensities of dark blue and transparent ring around colony were compared for activity measurement. Nutrient agar with 1% skim milk added was employed for measuing protease activity. 14 strains of Arthrobacter russicus with lipase activity and 19 strains of Chryserobacterium shigense with protease activities were found to be present. It was found that Acinetobacter genomospecies 10 (match %: 99.90) isolated from B region in fall was the most lipolytic species, whereas Serratia liquefaciens (match %: 99.39) isolated from the same region in spring was the most proteolytic species. Growth curve of Acinetobacte and Serratia liquefaciens was a typical sigmoidal form. Lipase activity increased with incubation time, but its activity began to drop at stationary to motality phase. Optimum condition for incubation time, pH and temperature for extracellular lipase from Acinetobacter genomospecies 10 (match %: 99.90) was 12 h, 8.5, and $45^{\circ}C$, respectively. Extracellular protease from Serratia liquefaciens (match %:99.39) had the same optimum incubation time and pH as extracellular lipase, but optimum temperature was $35^{\circ}C$.

• Korean Journal of Microbiology
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• v.46 no.1
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• pp.73-79
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• 2010

The Korea Polar Research Institute (KOPRI) has assembled a culture collection of cold-adapted bacterial strains from both the Arctic and Antarctic. To identify excellent protease-producers among the proteolytic bacterial collection (874 strains), 78 strains were selected in advance according to their relative activities and were subsequently re-examined for their extracellular protease activity on $0.1{\times}$ ZoBell plates supplemented with 1% skim milk at various temperatures. This rapid and direct screening method permitted the selection of a small group of 15 cold-adapted bacterial strains, belonging to either the genus Pseudoalteromonas (13 strains) or Flavobacterium (2 strains), that showed proteolytic activities at temperatures ranging between $5-15^{\circ}C$. The cold-active proteases from these strains were classified into four categories (serine protease, aspartic protease, cysteine protease, and metalloprotease) according to the extent of enzymatic inhibition by a class-specific protease inhibitor. Since highly active and/or cold-adapted proteases have the potential for industrial or commercial enzyme development, the protease-producing bacteria selected in this work will be studied as a valuable natural source of new proteases. Our results also highlight the relevance of the Antarctic for the isolation of protease-producing bacteria active at low temperatures.

• The Korean Journal of Zoology
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• v.37 no.3
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• pp.324-330
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• 1994

The multicatalvtic 205 proteasome consisting of 12-15 subunits of 22-35 kDa is the catalytic core of the ATP/ubiquitin-dependent 26S protease complex that also is comprised of multiple subunits of 22-110 KDa. In order to determine whether the proteolvtic activities change during muscle development, the enzyme preparations were obtained from 11-, 14- and 17-day old chick embryonic muscle using a BioGel A-1.5m column. The 26S complex preparation from 14- or 17-day old muscle hvdr olvz e d both N -s uccinvl- Le u- Le u -Val-Tvr-7- amido -4- methvlco umarin ( Suc- LLVY- AMC) and ubiquitin-Ivsozvme conjugates about 50% as well as that from 11-day old muscle. In addition, the activity of 20S proteBsome against Suc-LLVY-AMC also decreased by about 20-30%. However, the protein level of 265 complex remained constant during the entire development period, while that of 205 proteasome increased 5- to 6-fold, as analyzed by nondenaturins polyacrvlamide gel elenrophoresis followed by immunoblot analysis using the antibodies raised against the purified enzymes. Thus, the specific activity of 20S proteasome against the peptide must decrease rather dramatically during the muscle development. These results suggest that the development-dependent changes in the proteolytic activities of both 20S proteasome and 26S protease complect from embryonic muscle are due to alterations in the expression of certain subunits in the enzvmes that are responsible for their specific cataIVtic functions but not to overall changes in the enzyme amounts.

• Microbiology and Biotechnology Letters
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• v.2 no.2
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• pp.89-93
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• 1974

The various conditions of substituting wheat gluten for the bean, one of the raw materials for soy sauce manufacture, was studied by measuring the activities of the amylase and proteolytic enzyme of koji. It was found that substitute wheat gluten for up to 60％ of bean content (30％ of the total bran and wheat content) yielded good quality of soy sauce. By using more than 30％ of wheat gluten the availability of nitrogen of raw materials was decreased. This was attributed to the low enzyme activity in koji containing more than 30％ wheat gluten.

• Journal of Microbiology and Biotechnology
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• v.16 no.12
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• pp.1897-1903
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• 2006

In order to investigate the functions of the C-terminal region of chitinase ChiCW of Bacillus cereus 28-9, a C-terminal truncated enzyme, ChiCW$\Delta$FC, was expressed in Escherichia coli and purified to homogeneity for biochemical characterization. Compared with ChiCW, ChiCW$\Delta$FC exhibited higher chitinase activity at high temperature and pH, but expressed lower hydrolytic and binding activities toward insoluble substrates. In addition, kinetic properties indicated that ChiCW$\Delta$MC hydrolyzed oligomeric and polymeric substrates less efficiently than ChiCW. These results suggest that the C-terminal region of ChiCW plays important roles in substrate binding and hydrolysis of chitin. In addition, the biological meaning of C-terminal proteolytic modification of ChiCW is discussed.

• Microbiology and Biotechnology Letters
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• v.25 no.2
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• pp.189-196
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• 1997

A producer of inhibitor against ${\beta}-glucosidase$ of Penicillium sp.-L4 was screened from Actinomycetes, and the isolated strain was identified as Streptomyces sp. The inhibitor produced was very stable against heat, acidic and alkaline conditions, proteolytic and amylolytic enzymes. The inhibotor was purified from culture broth through activated carbon treatment, ultrafiltration, anion and cation exchange, activated carbon columm, acetone precipitation and preparative HPLC. It showed inhibitory activities against a variety of dissacharide hydrolyzing enzymes produced by P.sp.-L4, and the mode of inhibition was competitive. Its structure and molecular formular was elucidated by IR, $^1H\;and\;^{13}C$ NMR and FAB/Mass spectrometry, which was identified as 1-deoxynojirimycin (dNM). dNM showed inhibitory effects on the cell growth and hydrolytic enzyme action of P.sp.-L4 on agar plate and infected lemon peel.

• Biomedical Science Letters
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• v.10 no.4
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• pp.347-351
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• 2004

Maggot fibrolase (MsMg-1) was purified from the maggots of Mimela splendems using ammonium sulfate fractionation, DEAE Affi-gel affinity chromatography. This protease had a molecular weight of 85 kDa as determined by SDS-polyacrylarnide gel electrophoresis under reducing conditions. It showed strong proteolytic and fibrinolytic activities. The purified enzyme was strongly inhibited by phenylmethanesulfonyl fluoride, Mn/sup 2+/, and Zn/sup 2+/ but it was not by EDTA, EGT, Mg/sup 2+/, Ca/sup 2+/, and Li/sup 2+/ ions. In these experimental results, we have speculated that MsMg-1 is a serine protease with a strong fibrinolytic activity.

• Mycobiology
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• v.33 no.2
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• pp.121-123
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• 2005

Extracellular keratinase isolated from Aspergillus flavus K-03 was immobilized on calcium alginate. The properties and reaction activities of free and immobilized keratinase with calcium alginate were characterized. The immobilized keratinase showed proteolytic activity against soluble azo-casein and azo-keratin, and insoluble feather keratin. Heat stability and pH tolerance of keratinase were greatly enhanced by immobilization. It also displayed a higher level of heat stability and an increased tolerance toward alkaline pHs compared with free keratinase. During the durability test at $40^{\circ}C$, 48% of the original enzyme activity of the immobilized keratinase was remained after 7 days of incubation. The immobilized keratinase exhibited better stability, thus increasing its potential for use in industrial application.

• Journal of Life Science
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• v.8 no.6
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• pp.627-637
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• 1998

In order to utilize tuna pyloric caeca among fish intestines wasted when treated raw fish in fish processing manufactory, a crude enzyme with high proteolytic activity was extracted and its optimum condition were investigated. An immobilized enzymes also were prepared by adsorption method to enhance thermostability of the crude proteinase. The yield of the crude proteinase was approximately 2.7% on dry basis. The proteolytic activity for casein was 0.54 U/mg protein, for BTEE 1.10 U/mg protein, and for BAEE 2.69 U/mg protein. It was almost similar to that of the commercial trypsin purified. Optimum hydrolysis activity of the crude proteinase was about 80%, as the degree of hydrolysis for casein, at pH 10.0 and 45$^{\circ}C$ for 12 hrs. Also, when the crude proteinase was immobilized on DEAE-Cellulose and chitin, the residual activities remained after 7 days of pre-incubation time were maintained about 90% or more and their thermostabilities were enhanced by about 50%, compared with the native enzyme.