• BMB Reports
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• 제30권6호
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• pp.443-447
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• 1997

We have developed a novel method for assays of spermidine and spermine synthase (aminopropyltransferase) activities using antibody against 5'-deoxy-5'-methylthioadenosine (MTA). A new assay is reported here which is based on the observation that MTA is formed as a stoichiometric by-product of the spermidine and spermine synthases reactions. In order to determine MTA, a radioimmunoassay method with sensitivity and rapidity was used. (Lee and Cho, 1997). In this assay, adenine must be added in the reaction mixture, since it effectively inhibits the action of MTA phosphorylase by which MTA is metabolized. This assay is a improvement in term of sensitivity and time saving, compared to the currently used methods. It has a level of sensitivity (100 fmol) sufficient to monitor aminopropyltransferase activities in incubations containing as little as $10{\mu}g$ protein prepared from rat tissue homogenate. The results obtained showed that this method is particularly useful for cultured cells with low enzyme concentration. Moreover, this assay has the advantage which allows studies using alternative substrates (other amines). Spermidine synthase activity was high in rat liver, but low in rat kidney. The activity of spermine synthase was in most rat tissues very low as compared to that of spermidine synthase, but was high in brain.

• Asian Pacific Journal of Cancer Prevention
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• 제15권7호
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• pp.3005-3009
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• 2014

The aims of this study were to investigate the influence of ubiquitin- conjugating enzyme E2C (UBE2C) on biological behavior of lung cancer cells. Using MTT, flow cytometry and invasion assays, we detected UBE2C expression and evaluated its biological properties in these cells, including effects on proliferation, the cell cycle profile and invasive capability. Compared with control cells, the UBE2C transfected cells demonstrated increased cellular proliferation (p<0.05). UBE2C transfected cells also had a lower percentage in G1 phase and a higher percentage in S phase (p<0.05). Importantly, the UBE2C transfected cells had a notable enhancement of cell numbers penetrating the basement membrane compared with the control group (p<0.05). Ectopic up-regulation UBE2C promoted the growth of lung cancer cells in vivo. Furthermore, we found UBE2C increased the expression of cyclin D1 and MMP-2. These results show UBE2C may represent a potential therapeutic target for lung cancer.

• Clinical and Experimental Pediatrics
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• 제53권2호
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• pp.235-238
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• 2010

파브리병은 알파-갈락토시다아제(alpha galactosidase) A 효소에 위치하는 유전자 변이에 의한 글라이코스핑고리피드(glycospingolipid) 대사이상에 의한 질환으로 X-염색체와 연관된다. 말단 지각이상을 호소하던 12세 남자 환자와 증상이 미미한 그의 형에서 알파-갈락토시다아제 A 효소 활성도 측정과 유전자 변이 확인을 통하여 파브리병을 확진한 2례를 보고하고자 한다.

• BMB Reports
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• 제44권12호
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• pp.805-810
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• 2011

Methionine sulfoxide reductase A (MSRA) is a ubiquitous enzyme that has been demonstrated to reduce the S enantiomer of methionine sulfoxide (MetSO) to methionine (Met) and can protect cells against oxidative damage. In this study, we isolated a novel MSRA (SlMSRA2) from Micro-Tom (Solanum lycopersicum L. cv. Micro-Tom) and characterized it by subcloning the coding sequence into a pET expression system. Purified recombinant protein was assayed by HPLC after expression and refolding. This analysis revealed the absolute specificity for methionine-S-sulfoxide and the enzyme was able to convert both free and protein-bound MetSO to Met in the presence of DTT. In addition, the optimal pH, appropriate temperature, and $K_m$ and $K_{cat}$ values for MSRA2 were observed as 8.5, $25^{\circ}C$, $352{\pm}25\;{\mu}M$, and $0.066{\pm}0.009\;S^{-1}$, respectively. Disk inhibition and growth rate assays indicated that SlMSRA2 may play an essential function in protecting E. coli against oxidative damage.

• Mycobiology
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• 제47권4호
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• pp.430-440
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• 2019

Fungal contamination of built-in furniture is a frequent problem in Korea when new apartment is built. However, domestic information on the contaminating fungi is very limited. This study was conducted to isolate, identify and characterize the fungi of the problem in one of the apartment houses where the fungi were claimed in the built-in furniture before the house owner moves in. Fungi present in the furniture installed in a main room, dress room, and kitchen side were visually and microscopically confirmed and purely isolated on PDA. The isolated fungi were identified by analyzing the morphological characteristics and nucleotide sequence of the ITS, calmodulin gene, and TEF-1α gene. Aspergillus creber, A. niger, A. pseudoglacus, A. ruber, Cladosporium perangustum and Penicillium commune were identified. Four out of the six fungal species were positive for at least one enzyme in six kinds of extracellular enzyme assays. When these four species (A. creber, A. niger, C. perangustum and P. commune) were inoculated onto four kinds of wood chips of furniture materials, they were able to colonize all of the wood chips. Their settlement was better at 95% humidity condition than at 30% humidity condition. Among the four species, C. perangustum caused the darkest discoloration and secreted the most number of extracellular enzymes. The four species were re-isolated from the colonized wood chips and confirmed as the problematic fungi in the built-in furniture.

• Journal of Microbiology and Biotechnology
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• 제14권4호
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• pp.877-880
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• 2004

In this study, we determined the content of chitooligosaccharides (COS) in Korean commercial salt-fermented shrimps by competitive direct enzyme-linked immunosorbent assays (cdELISAs), using anti-COS mixture (COSM) antibody and COSM horseradish peroxidase (HRP) conjugate. When COS6 was spiked into salt-fermented shrimps at the level of $10-300\mu{g/g,}$ the average recovery was $120\pm19%$ ($mean\pmS.D.$). The COS contents of the 92 samples of Korean commercial salt-fermented shrimps collected during February 2000 and August 2002 were $36.3\pm20.7\mug$ COS6 equivalent/g (expressed as "$\mug/g$" hereafter). Among the samples, the COS contents of yuk-jeot ( $40.3 \pm 22.5 \mug/g, n=27$) and buksaewoojeot ($40.2 \pm 21.6 \mug/g, n=5$) were higher than the others. The COS contents of salt-fermented shrimps produced at Gwangcheon ($47.1 \pm 20.7 \mug/g, n=18$) and Gomso ($44.1 \pm 21.8 \mug/g, n=6$) areas were higher than those produced at the other areas. This is the first report to determine COS of salt-fermented shrimps by cdELISA.

• 한국임상수의학회지
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• 제34권3호
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• pp.185-188
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• 2017

This study assessed the seroprevalence of Babesia gibsoni in companion dogs in Korea by enzyme linked immunosorbent assay using recombinant BgTRAP antigen. Dogs were randomly selected from those admitted for various reasons to local private veterinary hospitals and the Animal Medical Center of Chonbuk National University. With the owners' permission, extra blood was drawn from each dog for serological assays. Of the 188 selected dogs, seven (3.72%) were positive for B. gibsoni, including six of 167 (3.59%) indoor and one of 12 (8.33%) outdoor dogs. Of the seven dogs positive for B. gibsoni, four were aged > 10 years, two were < 1 year, and one was between 1 and 10 years; and two were Yorkshires and one each was Shih-tzu, Maltese, Pekinese, beagle and mixed. Concurrent diseases or chief complaints were anemia in two dogs, both of which had a history of confirmed babesiosis by polymerase chain reaction, and non-anemic diseases in five. Geographically, four dogs were from Jeonbuk/Jeonju, and one each from Seoul, Gyounggi-do, and Jeonnam/Gwangju. To our knowledge, this is the first report of companion dogs in Korea being seropositive for B. gibsoni. Serologic screening of subclinical or carrier dogs can detect this potentially dangerous disease and assess its epidemiology.

• Biomolecules & Therapeutics
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• 제3권4호
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• pp.245-250
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• 1995

The carbon tetrachloride($CCl_4$) has been demonstrated to have a hepatotoxic effect in human or many other species. To investigate the enzyme induction of mixed function oxygenases in liver of male Sprague-Dawley rats a single 0.1, 0.5 mι/kg dose of carbon tetrachloride were given. At 24 hr after a single dose of 0.1 mι CC1$_4$/kg weight, methanol extract of Hedera rhombea leaves was administered with 100, 500 mg/kg weight. Assays of 7-ethoxyresorufin-Ο-deethylation(EROD),7-benzyloxyresorufin-Ο-deathylation(BROD),4-nitro-phenol-UDP-glucuronosyltransferase(UDPGT), Western blot and RNA slot blot were used as representatives of the activities of cytochrome P-450 enzymes. The change of the activity of CYP1A1 form measured by EROD assay and Western analysis using 1-7-1 monoclonal antibody was not observed. The activity CYP2B1 form by BROD assay and using 2-66-3 monoclonal antibody was remarkably increased. Elevated level of CYP2B1 mRNA was shown by slot hybridization with 2B1-specific probe. Administration of methanol extract of Hedera rhombea leaves reversed the enzyme activity and the level of mRNA, which suggest the chemoprotective effect of methanol extracts of Hedera rhombea leaves to carbon tetrachloride hepatotoxlcity.

• 생명과학회지
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• 제16권2호
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• pp.274-281
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• 2006

흑두청국으로부터 분리된 혈전용해력이 우수한 Bacillus subtilis BB-1(KFCC 11344P)으로부터 혈전용해효소 유전자를 PCR법에 의해 크로닝하였고 이를 BCF-1으로 명명하였다. BCF-1의 DNA 염기서열결정 결과 1,145 bp 크기의 혈전용해 효소로, 일본의 natto로부터 분리된 nattokinase 유전자와 99%의 상동성을 보임을 확인하였다. 혈전용해효소 유전자의 발현을 위하여 Bacillus 발현계인 Bacillus-E. coli의 shuttle vector인 pEB vector에 크로닝 하고 host로서 B. subtilis 168에 형질전환시켜 대량 발현시켰다. 생산된 혈전용해효소의 최 적활성 pH와 온도는 7.0과 $35^{\circ}C$로 확인되었다, 기질에 대한 분해양상을 조사한 결과 fibrin에서만 특이적으로 강한 분해가 일어났으며, skim milk에서 아주 약한 분해능을 보였으나 blood agar, gelatin, casein에서는 전혀 분해능을 보이지 않았다. 특히 blood agar plate에서 분해능이 없는 것으로 보아 혈액 내에서의 적혈구 파괴현상과 같은 부작용에 대한 위험을 배제할 수 있을 것으로 사료된다. BCF-1에 의해 생산된 혈전용해효소는 fibrin 특이적으로 활성을 나타냄을 확인할 수 있으며, 이는 임상적이나 산업적으로 적용하였을 때 부작용에 대한 위험적인 문제는 배제될 수 있으리라 생각된다.

• 한국축산식품학회지
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• 제38권1호
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• pp.135-142
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• 2018

Recently, research on the processing of raw functional materials with the aim of improving various physiological activities has been conducted. In this study, we investigated the antioxidant activity of royal jelly (RJ) hydrolysates obtained from three commercial proteases. Enzyme-treated royal jelly (ERJ), in which the RJ hydrolysates were converted into easy-to-absorb shorter chain monomers through the removal of two known allergen proteins, showed no difference in the content of (E)-10-hydroxydec-2-enoicacid (10-HDA) or the freshness parameter and showed a significant increase in total free amino acid content. The antioxidant activity of ERJ was determined by 1,1-diphenyl-2-picrylhydrazyl (DPPH) and chemical assays. The ERJ showed about 80% DPPH-radical scavenging activity at same concentration of ascorbic acid. The antioxidant effect of ERJ was confirmed to be due to reduction of intracellular reactive oxidative species (ROS) and nitric oxide (NO) production in LPS-treated macrophages. Moreover, ERJ significantly increased the activity of the antioxidant enzyme superoxide dismutase (SOD) and the level of the antioxidant glutathione (GSH) in a dose-dependent manner. Interestingly, these antioxidant activities of ERJ were stronger than those of non-treated RJ. These findings indicate that ERJ has high potential as an antioxidant agent for use in human and animal diets.