• Parasites, Hosts and Diseases
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• v.52 no.1
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• pp.41-46
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• 2014

The mature domain of a cysteine protease of Spirometra erinacei plerocercoid larva (i.e., sparganum) was expressed in Escherichia coli, and its value as an antigen for the serodiagnosis of sparganosis was investigated. The recombinant protein (rSepCp-1) has the molecular weight of 23.4 kDa, and strongly reacted with the sparganum positive human or mice sera but not with negative sera by immunoblotting. ELISA with rSepCp-1 protein or sparganum crude antigen (SeC) was evaluated for the serodiagnosis of sparganosis using patient's sera. The sensitivity and specificity of ELISA using rSepCp-1 protein were 95.0% (19/20) and 99.1% (111/112), respectively. In contrast, the sensitivity and specificity of ELISA with SeC were 100% (20/20) and 96.4% (108/112), respectively. Moreover, in experimentally infected mice, the sensitivity and specificity of both ELISA assays were 100% for the detection of anti-sparganum IgG. It is suggested that the rSepCp-1 protein-based ELISA could provide a highly sensitive and specific assay for the diagnosis of sparganosis.

• Journal of Plant Biology
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• v.38 no.1
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• pp.1-10
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• 1995

Photoinhibition and its recovery of spectrally adapted Anacystis nidulans were studied. Phycocyanin and Chl content and phycocyanin/Chl ratio were increased in cells grown under blue-green light compared with those grown in white light. Photosynthetic activities of white light and blue-green light grown cells were reduced by 50% after 15 min and 10 min of photoinhibitory light treatment (1.2 mmol·m-2s-1), respectively, largely due to the decline of PSII activities. However, their activities were recovered fully after 30 min incubation under weak light. Treatment of rifampicin and chloramphenicol magnified the photoinhibitory effects and suppressed the recovery with disappearance of susceptibility to photoinhibition and delayed the recovery process, indicating no significant differences in phosphorylation, dephosphorylation and protease activity between two cells. Therefore, it is suggested that the increased sensitivity of blue-green adapted cells might be attributed to the decline of protein synthesis, and phosphorylation-dephosphorylation of protein and protease activity might be involved in the recovery process.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.21 no.2
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• pp.71-79
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• 1988

For the purpose of producing low salt fermented anchovy by accelerated method with a strong proteolytic bacteria, in this study, a strong proteolytic bacterium was isolated from low salt fermented anchovy and its bacteriological characteristics and properties of protease were experimented. The results obtained were as fellows : three proteolytic bacteria, Aeromonas anaerogenes Barillus subtilis and Staphylococcus saprophyticus were isolated from low salt fermented anchovy($4\%\;of\;salt,\;4\%\;of\;KCl,\;0.5\%\;of\;lactic\;acid,\;6\%$of sorbitol and $4\%$ of alcohol extract of red pepper) after 40 days fermentation. Among these strains, which grow best at $30^{\circ}C$, pH 7.0, B. subtilis was found the best proteolytic strain and benefit for industrial use as shown $0.95\;hr^{-1}$ of specific growth rate, $89{\mu}g-Tyr/hr.ml$ of maximum activity after 12 hrs culture in TPY broth. The protease produced by by B. subtilis showed maximum activity at $35^{\circ}C$, pH 7.0, and molecular weight was estimated to be 23,000 by Sephadex G-100 filtration, and it was supposed to be a kind of metal chelator sensitive neutral protease from the results of strong sensitivity against EDTA, o-phenanthroline and metal ions such as $Cu^{2+},\;Ni^{2+},\;Fe^{2+}.Km$ value of that by method of Lineweaver-Burk was determinded to be $0.73\%$ for casein as a substrate.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.22 no.5
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• pp.363-369
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• 1989

In this study, the biochemical characterization of strong proteolytic bacteria and their extracellular pretense isolated from fermented fish paste were experimented for the purpose of industrial large scale-production by accelerated fermentation. The results obtained were as follows: Among 4 strains isolated from fermented fish paste, B. subtilis p-4 and B. licheniformis p-5, which grow well at $40^{\circ}C$, pH 7.0 and $1\%$ of salt contents, were the best proteolytic bacteria and were shown $0.48hr^{-1}$, $0.49hr^{-1}$ of specific growth rate in TPY medium, respectively. Maximum enzyme activity of B. subtilis p-4 was 335n mole-Tyr/min.ml after 30 hrs and that of B. licheniformis p-5 was 300n mole-Tyr/min.ml after 28 hrs of shaking culture. Purified pretense produced by B. subtilis p-4 and B. licheniformis p-5 showed maximum activity at $50^{\circ}C$, pH 7.0 and molecular weight were estimated to be 18,000, 30,000 by sephadex G-100 gel filtration, respectively. These were supposed to be a kind of metal chelator sensitive neutral pretense from the result of high sensitivity against EDTA, o-phe-nanthroline and metal ions such as $Cu^{2+},\;Ni^{2+},\;Zn{2+}$.

• Microbiology and Biotechnology Letters
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• v.21 no.2
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• pp.125-131
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• 1993

The s-type pyocin, one kind of bacteriocins, is a bactericidal substabce of protein nature produced by certain Pseudomonas aeruginosa and is active against some other strains of the same or closely related species. Among many Pseudomonas aeruginosa strains collected from the patients at the Hospitals in Seoul and Taegu cities, some Pseudomonas aeruginosa pyocinogeny were determined by pyocin typing. As a result, Pseudomonas aeruginosa 90-2-2205 was selected as the S-type pyocin producing microorganism due to its highest antimicrobial and protease sensitivity.

• Korean Journal of Microbiology
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• v.30 no.4
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• pp.269-277
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• 1992

Several insertion mutants of Escherichia coli in the ant gene, coding for $Na^{+}$ $H^{+}$ antiport activity, showed littel, if any, reduction in the antiport activity. $Na^{+}$ dependent transport activity also remained at wild type level. These facts led to the idea that E. coli has evolved at least two distinct systems for extrusion of $Na^{+}$ The antiport activities were studied under various conditions to reveal different properties of these systems. For convenience these activities are referred to as major and minor activities. The distinguishing properties of the two systems include : kinetics (Km, Vm) at pH 7.8, competition pattern between $Na^{+}$ and Li$^{+}$ , pH profiles, pattern of the change in kinetic parameters as a function of pH, and sensitivity to protease, chemicals and heat.

• Mycobiology
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• v.50 no.5
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• pp.382-388
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• 2022

White mold (or Sclerotinia stem rot), caused by Sclerotinia species, is a major air, soil, or seed-transmitted disease affecting numerous crops and wild plants. Microscopic or culture-based methods currently available for their detection and identification are time-consuming, laborious, and often erroneous. Therefore, we developed a multiplex quantitative PCR (qPCR) assay for the discrimination, detection, and quantification of DNA collected from each of the three economically relevant Sclerotinia species, namely, S. sclerotiorum, S. minor, and S. nivalis. TaqMan primer/probe combinations specific for each Sclerotinia species were designed based on the gene sequences encoding aspartyl protease. High specificity and sensitivity of each probe were confirmed for sclerotium and soil samples, as well as pure cultures, using simplex and multiplex qPCRs. This multiplex assay could be helpful in detecting and quantifying specific species of Sclerotinia, and therefore, may be valuable for disease diagnosis, forecasting, and management.

• Asian-Australasian Journal of Animal Sciences
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• v.28 no.5
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• pp.620-631
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• 2015

A total of 59 lactic acid bacteria (LAB) strains were isolated from corn stover silage. According to phenotypic and chemotaxonomic characteristics, 16S ribosomal DNA (rDNA) sequences and recA gene polymerase chain reaction amplification, these LAB isolates were identified as five species: Lactobacillus (L.) plantarum subsp. plantarum, Pediococcus pentosaceus, Enterococcus mundtii, Weissella cibaria and Leuconostoc pseudomesenteroides, respectively. Those strains were also screened for antimicrobial activity using a dual-culture agar plate assay. Based on excluding the effects of organic acids and hydrogen peroxide, two L. plantarum subsp. plantarum strains ZZU 203 and 204, which strongly inhibited Salmonella enterica ATCC $43971^T$, Micrococcus luteus ATCC $4698^T$ and Escherichia coli ATCC $11775^T$ were selected for further research on sensitivity of the antimicrobial substance to heat, pH and protease. Cell-free culture supernatants of the two strains exhibited strong heat stability (60 min at $100^{\circ}C$), but the antimicrobial activity was eliminated after treatment at $121^{\circ}C$ for 15 min. The antimicrobial substance remained active under acidic condition (pH 2.0 to 6.0), but became inactive under neutral and alkaline condition (pH 7.0 to 9.0). In addition, the antimicrobial activities of these two strains decreased remarkably after digestion by protease K. These results preliminarily suggest that the desirable antimicrobial activity of strains ZZU 203 and 204 is the result of the production of a bacteriocin-like substance, and these two strains with antimicrobial activity could be used as silage additives to inhibit proliferation of unwanted microorganism during ensiling and preserve nutrients of silage. The nature of the antimicrobial substances is being investigated in our laboratory.

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.10
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• pp.1709-1714
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• 2004

The changes of microflora and enzyme activities in Alaska pollack sikhae were evaluated in 3 different temperature conditions, 5$^{\circ}C$, 2$0^{\circ}C$ and alternating temperature (stored at 5$^{\circ}C$ after 10 days of fermentation at 2$0^{\circ}C$), respectively. The number of proteolytic bacteria and 2 lactic acid bacteria including Lactobacillus sp. and Pediococcus sp. increased rapidly up to 10 days and composed major portion of total viable cell (TVC) in sikhae fermented at 2$0^{\circ}C$, whereas those of TVC were occupied by Lactobacillus sp., Pediococcus sp. and yeast after 10 days of fermentation. The major species of microflora in sikhae fermented at alternating temperature were, composed of Lactobacillus, Pediococcus and Streptococcus after 10 days of fermentation. Especially, Leuconostoc sp. was kept up to 27 days at 5$^{\circ}C$ than other temperature conditions (16 days). The activities of protease and lipase in acidic region (pH 3.0) were higher at 2$0^{\circ}C$ than at 5$^{\circ}C$ due to sensitivity of temperature, although those of protease and lipase in neutral region (pH 7.0) were not found any differences in both temperatures. Changing temperature condition from 2$0^{\circ}C$ to 5$^{\circ}C$ in alternating temperature inactivated protease activity, whereas lipase activity was still maintained during fermentation.

• The Korean Journal of Cytopathology
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• v.11 no.2
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• pp.75-81
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• 2000

Cathepsin D is a protease which is known to facilitate invasion and metastasis of breast carcinoma. Overexpression of cathepsin D is associated with poor clinical outcome and biologic aggressiveness of the breast cancer. We underwent immunocytochemical assay(ICA) for cathepsin D in fine needle aspiration cytology(FNAC) specimens from the breast carcinoma and benign breast diseases. In FNAC specimens cathepsin D was expressed in 21(42.9%) out of 49 cases of invasive ductal carcinoma, whereas negative result was observed in all 15 cases of benign breast diseases including 7 fibroadenomas, 6 fibrocystic diseases, and 2 benign ductal hyperplasias. Among the 11 FNAC specimens from ductal carcinoma in situ(DCIS), cathepsin D was expressed in 3 cases(27.3%). In FNAC specimens immunocytochemistry for cathepsin D showed positive result in 24 out of 60 carcinomas(sensitivity, 40%) and negative result in 15 out of all 15 benign breast diseases(specificity, 100%). No significant correlation was noted between cathepsin D expression in FNAC specimen and clinicohistological characteristics of the breast carcinoma, such as hormone receptors and cell differentiation. In conclusion, ICA of cathepsin D in FNAC specimens thought to be a good adjunct to differentiate malignancy from benign breast diseases.