• KSBB Journal
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• 제24권3호
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• pp.253-258
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• 2009

본 연구에서는 ELISA와 면역-크로마토그래피 스트립기술을 결합하여 E. coli O157 : H7을 탐지할 수 있는 면역스트립 센서를 제작하기 위한 제작조건의 최적화 연구를 수행하였다. 포획항체 농도, 탐지항체 농도, 완충용액 첨가제 조성의 면역스트립 제작 또는 운전인자들의 최적화 조건을 결정하였다. 포획항체의 농도는 1 mg/mL를 최적 조건으로 선정하였고, 탐지항체의 최적 농도도 1 mg/mL로 결정하였다. 비특이적 결합을 방지하기 위한 시료 희석용 완충용액의 첨가제 조성으로는 0.5% Tween 20와 3% BSA 혼합 사용을 선정하였다.

• 대한수의학회지
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• 제62권1호
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• pp.4.1-4.5
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• 2022

This paper describes a fatal case of pneumonia in a 14-day-old dog caused by extraintestinal pathogenic Escherichia coli (ExPEC). The necropsy showed that almost all of left lobes of the lungs had dark-red consolidation. A histopathology examination revealed moderate acute fibrino-hemorrhagic necrotizing pneumonia with intralesional bacterial colonies. Non-suppurative epicarditis, congestion in the liver, and necrosis in the white pulp of the spleen also were found. E. coli with cytotoxic necrotizing factor 1 and α-hemolysin was isolated from the lung. This case was confirmed to have fatal pneumonia caused by ExPEC that led to a systemic infection.

• 한국식품과학회지
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• 제31권6호
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• pp.1577-1582
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• 1999

자체 제작한 고전압 펄스 전기장 장치를 이용하여 당근주스의 주요 변패 미생물인 대장균(Escherichia coli)의 불활성에 미치는 전기장 세기, 처리시간, 온도 등을 관찰하고 품질변화에 관하여 연구하였다. E. coli의 불활성에 필요한 최소 전기장세기인 임계전기장 세기(Ec)는 11.74 kV/cm, 최소처리시간(tc)는 $3.6{\mu}s$이었다. 사멸효과는 전기장의 세기가 클수록, 처리시간이 길수록 그리고 처리온도가 높을수록 증가하였다. 당근주스의 pH, 가용성 고형분 $(^{\circ}Brix)$, 적정산도는 거의 변화가 없었다. 색도는 50^{\circ}$ 열병합 PEF처리 주스만이 약간 감소하는 경향을 보였지만 다른 처리구들은 변화가 거의 없었다. ${\alpha},\;{\beta}-carotene$의 함량도 거의 변화가 없었다.

• 한국축산식품학회지
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• 제30권2호
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• pp.179-184
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• 2010

Escherichia coli O157:H7 is a food-borne pathogen that causes bloody diarrhea, hemorrhagic colitis, and hemolytic uremic syndrome (HUS). We compared three selective media and evaluated the performance of immunomagnetic separation (IMS) for the detection of low levels of E. coli O157:H7 in ground beef and radish sprouts with different levels of background flora. Bulk food samples (500 g for each trial) were artificially inoculated with nalidixic acid-resistant E. coli O157:H7 at the lowest dose that would generate 20 partial-positive samples of 25 g each. All samples were homogenized in mTSB (225 mL) and incubated overnight at $37^{\circ}C$. IMS was performed using the enriched mTSB samples (1 mL) along with conventional spreads plated onto three different selective media: Sorbitol MacConkey agar (SMAC), Sorbitol MacConkey agar with cefixime and tellulite (CT-SMAC), and Sorbitol MacConkey agar with nalidixic acid (NAL-SMAC) as the gold standard. Two suspicious colonies from each medium were selected and confirmed usinga serological test after transfer to tryptic soy broth with yeast extract (TSAYE). CT-SMAC was better than SMAC for detecting E. coli O157:H7 in all food types. Although there was no statistical difference in the number of positive samples when using IMS vs. non-IMS techniques, more positive samples were detected when IMS was used in both ground beef and radish sprouts. It appears that the improvement was more significant in radish sprouts, which had a higher level of background flora than ground beef. The results also suggest that the combination of CT-SMAC and IMS is sufficient to recover low levels of E. coli O157:H7 in high background flora food samples.

• Journal of Microbiology and Biotechnology
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• 제26권4호
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• pp.710-716
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• 2016

Gamma-aminobutyric acid (GABA) is a non-protein amino acid, which is an important inhibitor of neurotransmission in the human brain. GABA is also used as the precursor of biopolymer Nylon-4 production. In this study, the carbon flux from the tricarboxylic acid cycle was directed to the GABA shunt pathway for the production of GABA from glucose. The GABA shunt enzymes succinate-semialdehyde dehydrogenase (GabD) and GABA aminotransferase (GabT) were co-localized along with the GABA transporter (GadC) by using a synthetic scaffold complex. The co-localized enzyme scaffold complex produced 0.71 g/l of GABA from 10 g/l of glucose. Inactivation of competing metabolic pathways in mutant E. coli strains XBM1 and XBM6 increased GABA production 13% to reach 0.80 g/l GABA by the enzymes co-localized and expressed in the mutant strains. The recombinant E. coli system developed in this study demonstrated the possibility of the pathway of the GABA shunt as a novel GABA production pathway.

• Biotechnology and Bioprocess Engineering:BBE
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• 제8권4호
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• pp.227-232
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• 2003

A self-assembled monolayer of protein G was fabricated to develop an immunosensor based on surface plasmon resonance (SPR), thereby improving the performance of the antibodybased biosensor through immobilizing the antibody molecules (lgG). As such, 11-mercaptoundecanoic acid (11-MUA) was adsorbed on a gold (Au) support, while the non-reactive hydrophilic surface was changed through substituting the carboxylic acid group (-COOH) in the 11-MUA molecule using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrocholide (EDAC). The formation of the self-assembled protein G layer on the Au substrate and binding of the antibody and antigen were investigated using SPR spectroscopy, while the surface topographies of the fabricated thin films were analyzed using atomic force microscopy (AFM). A fabricated monoclonal antibody (Mab) layer was applied for detecting E. coli O157:H7. As a result, a linear relationship was achieved between the pathogen concentration and the SPR angle shift, plus the detection limit was enhanced up to 10$^2$ CFU/mL.

• 한국식품과학회지
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• 제52권1호
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• pp.103-108
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• 2020

본 연구는 식품에서 문제가 되는 시가독소생성 대장균(STEC)을 박테리오파지 증폭 기법을 통해 검출하고자 시가독소 생성 대장균에 대한 박테리오파지를 분리하였고 분리된 4종의 파지와 기 분리된 2종의 박테리오파지를 혼합하여 사용하였다. 분리된 박테리오파지는 형태학적 특성 및 제한효소 절단 패턴 등을 통해서 동정하였다. 5종의 파지는 E. coli O157:H7 및 non-O157 시가독소 생성 대장균을 모두 저해하는 특징을 가지는 것으로 나타났다. 박테리오파지 증폭 기법에서 중요한 단계인 세균에 감염되지 않은 박테리오파지를 제거하기 위해 10% (v/v) ferrous ammonium sulfate (FAS)을 사용하였으며 약 7-9 log PFU/mL 수준의 박테리오파지를 10분 내로 제거하는 것을 확인하였다. 시가독소 생성 대장균인 E. coli NCCP 13937을 검출하기 위해서는 약 6 log PFU/mL 이상의 박테리오파지 혼합액의 농도 및 약 4-5 log CFU/mL 이상의 목표 균주가 필요한 것으로 나타났다. 이러한 조건을 바탕으로 실제 판매되고 있는 신선식품에서 시가독소생성 대장균을 검출한 결과, 5시간 이내에 증폭된 약 2-3 log PFU/mL의 plaque를 통해 검출이 가능한 것을 확인하였다. 따라서 본 연구를 통해 박테리오파지 혼합액을 이용한 증폭 기법을 통해 시가독소 생성 대장균의 오염 여부를 보다 효율적으로 확인할 수 있음을 보여주었고 이를 적용한 제품을 개발하여 검출 단계의 간편화가 가능할 것으로 판단된다.

• 생명과학회지
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• 제8권1호
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• pp.40-50
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• 1998

Staphylococuus aureus and Escherichia coli is increasingly responsible for outbreaks of nosocomial infection around the world. Because serious infections due to these organisms currently necessiate use of non-$\beta$-lactam antimicrobial therapy and because strains is ofen resistant to many antimicrobial agents, infections with this organism are difficult to treat. Isolated strains from post operaton wounds of PNU hospital patient were tested for the antimicrobial susceptibility, resistant pattern and combined action to the 6 antibiotics. The minimal inhibitory concentraction of each antibiotic anc antibiotics combining in various ratios were measured by checkerboard dilution method. the synergism was determined through calculating the fractional inhibitory concentraction index (FICI). In case of S. aureus, 15 strains was shown to be highly sensitive to streptomycin and 13 strains to cephalothin. In case of E. coli, it is excellent senstitive 16 strains, sensitive 4 strains on cefoperazone, as like S. aureus, and thus the sensitive is most to be 66%. As the result of gaining MIC from S. aureus upon agar dilution method, MIC$_{50}$ was 8$\mu$g/ml, MIC$_{90}$ was 16$\mu$g/ml and thus the streptomycine is shown to be lowest. In case of E, coli, S. MIC$_{50}$ was 4$\mu$g/ml, MIC$_{90}$ was 16$\mu$g/ml, in streptomycin and thus is shown to be lower than S. aureus. As the result of comparing the resistance aspect of combining the antibiotics on S. aureus and E. coli, the resistant strain can be known to be reduced to the large range more than each 40% than combining with only aminoglycoside-series or cephalosporine-series. As the result of combining aminoglycoside-series, streptomycin and cephalothin or cefuroxime sensitive to S. aureus and E. coli in the above mentioned results, the increase or imporovement of effect is over 73% and 80%, respectively, thus the case od combining 2 antibiotics is shown to be better in the effect. Isolated strains from operating wounds were for the antimicrobial susceptibility. In case of S. aureus 15 strains was shown to be sensitive very much on streptomycin. In case of E. coli it is excellent sensitive 16 strains. As the results of combining aminoglycosides-series, streptomycin and cephalosporine series, cephalothin and cefuroxime, the increase or improvement of effect is over 73%, thus case of combining 2 antibiotics is shown to be better in the effect.

• Journal of Microbiology and Biotechnology
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• 제20권6호
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• pp.1018-1021
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• 2010

Metallic and non-metallic isomerases can be used to produce commercially important monosaccharides. To determine which category of isomerase is more suitable as a template for directed evolution to improve enzymes for galactose isomerization, L-arabinose isomerase from Escherichia coli (ECAI; E.C. 5.3.1.4) and tagatose-6-phosphate isomerase from Staphylococcus aureus (SATI; E.C. 5.3.1.26) were chosen as models of a metallic and non-metallic isomerase, respectively. Random mutations were introduced into the genes encoding ECAI and SATI at the same rate, resulting in the generation of 515 mutants of each isomerase. The isomerization activity of each of the mutants toward a non-natural substrate (galactose) was then measured. With an average mutation rate of 0.2 mutations/kb, 47.5% of the mutated ECAIs showed an increase in activity compared with wild-type ECAI, and the remaining 52.5% showed a decrease in activity. Among the mutated SATIs, 58.6% showed an increase in activity, whereas 41.4% showed a decrease in activity. Mutant clones showing a significant change in relative activity were sequenced and specific increases in activity were measured. The maximum increase in activity achieved by mutation of ECAI was 130%, and that for SATI was 190%. Based on these results, the characteristics of the different isomerases are discussed in terms of their usefulness for directed evolution of non-natural substrate isomerization.

• Asian-Australasian Journal of Animal Sciences
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• 제18권9호
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• pp.1336-1342
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• 2005

The present research work was conducted to evaluate the beneficial effects as well as the safety aspects of lactobacilli as probiotic. Lactobacilli were isolated from poultry faecal samples, feed samples and from some known preparations procured from poultry feed manufacturers. L. acidophilus and L. sporogenes were tested for the antibacterial activity against four poultry pathogens viz. Escherichia coli, Salmonella spp., Proteus spp. and Pseudomonas aeruginosa. Cell free supernatant (CFS) of L. acidophilus exhibited significantly higher antibacterial activity against Salmonella spp. at original pH (4.50${\pm}$0.02). At the adjusted pH (6.50${\pm}$0.02) significantly higher antibacterial activity was recorded against indicator organism except for P. aeruginosa. Likewise, L. sporogenes exhibited similar antibacterial activity at original as well as adjusted pH except for E. coli. Antibacterial activity against E. coli was significantly higher at adjusted pH than at original pH of CFS. The competitive exclusion of E. coli by lactobacilli over the intestinal epithelial cells (IEC) was checked. L. acidophilus strain I, which was of poultry origin, exhibited maximum attachment over IEC as compared to other three strains of non-poultry origin viz. L. acidophilus strain II, L. sporogenes strain I and II. Overall, L. acidophilus exhibited higher competitive exclusion as compared to L. sporogenes. All the lactobacilli of poultry origin were most sensitive to penicillin G, amoxycillin, ampicillin and chloramphenicol, least sensitive to sulphamethizole, ciprofloxacin, neomycin, norfloxacin and pefloxacin and resistant to metronidazole and nalidixic acid. The isolates from probiotic preparations were most sensitive to ampicillin, amoxycillin and tetracycline, least sensitive to sulphamethizole, norfloxacin, neomycin and ceftriazone and resistant to nalidixic acid and metronidazole. Eight of the multiple drug resistant lactobacilli isolates were studied for the presence of plasmids. Plasmids could be extracted from six isolates of lactobacilli. These plasmids could be responsible for bacteriocin production or for antibiotic resistance of the strains. The lactobacilli need further studies regarding their safety for use in the probiotic preparations.