• 한국미생물·생명공학회지
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• 제20권2호
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• pp.136-142
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• 1992

토양 분리균인 B.stearothermophilus chromosome의 유전자 은행으로부터 E.coli HB101 균주에 $\beta$-D-xylosidase 생산능력을 갖게하는 5.4Kb와 6.4Kb의 두 DNA 절편을 분리, pBR322에 크로닝하여 각각 pMG01과 pMG02의 재조합 플라스미드를 얻었다. 상기 두 B.stearothermophilus DNA 절편의 restriction map을 작성하고 이것을 기초로 하여 $\beta$-D-xylosidase 유전자인자의 위치를 확인함과 동시에 pUC18에 subcloning하여 각각 2.2kb와 1.0kb의 DNA 단편이 삽입된 $\beta$-D-xylosidase 양성의 pMG1와 pMG2를 분리하였다.

• 농업과학연구
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• 제47권3호
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• pp.645-655
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• 2020

Bacteria are a very common cause of food poisoning. Moreover, bacteria form biofilms to protect themselves from harsh environments. Conventional detection methods for foodborne bacterial pathogens including the plate count method, enzyme-linked immunosorbent assays (ELISA), and polymerase chain reaction (PCR) assays require a lot of time and effort. Hyperspectral imaging has been used for food safety because of its non-destructive and real-time detection capability. This study assessed the feasibility of using hyperspectral imaging and machine learning techniques to detect biofilms formed by Escherichia coli. E. coli was cultured on a high-density polyethylene (HDPE) coupon, which is a main material of food processing facilities. Hyperspectral fluorescence images were acquired from 420 to 730 nm and analyzed by a single wavelength method and machine learning techniques to determine whether an E. coli culture was present. The prediction accuracy of a biofilm by the single wavelength method was 84.69%. The prediction accuracy by the machine learning techniques were 87.49, 91.16, 86.61, and 86.80% for decision tree (DT), k-nearest neighbor (k-NN), linear discriminant analysis (LDA), and partial least squares-discriminant analysis (PLS-DA), respectively. This result shows the possibility of using machine learning techniques, especially the k-NN model, to effectively detect bacterial pathogens and confirm food poisoning through hyperspectral images.

• Journal of Animal Science and Technology
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• 제65권3호
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• pp.611-626
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• 2023

Escherichia coli (E. coli) and Salmonella enterica (SE) infections in pigs are major source associated with enteric disease such as post weaning diarrhea. The aim of this study was to investigate the effects of Pediococcus pentosaceus in weaned piglets challenged with pathogen bacteria. In Experiment.1 90 weaned piglets with initial body weights of 8.53 ± 0.34 kg were assigned to 15 treatments for 2 weeks. The experiments were conducted two trials in a 2 × 5 factorial arrangement of treatments consisting of two levels of challenge (challenge and non-challenge) with E. coli and SE, respectively and five levels of probiotics (Control, Lactobacillus plantarum [LA], Pediococcus pentosaceus SMFM2016-WK1 [38W], Pediococcus acidilactici K [PK], Lactobacillus reuteri PF30 [PF30]). In Experiment.2 a total of 30 weaned pigs (initial body weight of 9.84 ± 0.85 kg) were used in 4 weeks experiment. Pigs were allocated to 5 groups in a randomized complete way with 2 pens per group and 3 pigs per pen. Supplementation of LA and 38W improved (p < 0.05) growth performance, intestinal pathogen bacteria count, fecal noxious odor and diarrhea incidence. In conclusion, supplementation of 38W strains isolated from white kimchi can act as probiotics by inhibiting E. coli and SE.

• Journal of Microbiology and Biotechnology
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• 제26권2호
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• pp.356-363
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• 2016

Human papillomavirus (HPV), a non-enveloped, double-stranded DNA tumor virus, is a primary etiological agent of cervical cancer development. As a potential tool for prophylactic vaccination, the development of virus-like particles (VLPs) containing the HPV16 L1 capsid protein is highly desired. In this study, we developed a high-level expression system of the HPV16 L1 in Escherichia coli for the purpose of VLP development. The native gene of HPV16 L1 has many rare codons that cause the early termination of translation and result in the production of truncated forms. First, we optimized the codon of the HPV16 L1 gene to the preferable codons of E. coli, and we succeeded in producing the full-size HPV16 L1 protein without early termination. Next, to find the best host for the production of HPV16 L1, we examined a total of eight E. coli strains, and E. coli BL21(DE3) with the highest yield among the strains was selected. With the selected host-vector system, we did a fed-batch cultivation in a lab-scale bioreactor. Two different feeding solutions (complex and defined feeding solutions) were examined and, when the complex feeding solution was used, a 6-fold higher production yield (4.6 g/l) was obtained compared with that with the defined feeding solution.

• Journal of Microbiology and Biotechnology
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• 제31권9호
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• pp.1191-1199
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• 2021

Enteropathogenic Escherichia coli (EPEC), which belongs to the attaching and effacing diarrheagenic E. coli strains, is a major causative agent of life-threatening diarrhea in infants in developing countries. Most EPEC isolates correspond to certain O serotypes; however, many strains are non-typeable. Two EPEC strains, EPEC001 and EPEC080, which could not be serotyped during routine detection, were isolated. In this study, we conducted an in-depth characterization of their putative O-antigen gene clusters (O-AGCs) and also performed constructed mutagenesis of the O-AGCs for functional analysis of O-antigen (OAg) synthesis. Sequence analysis revealed that the occurrence of O-AGCs in EPEC001 and E. coli O132 may be mediated by recombination between them, and EPEC080 and E. coli O2/O50 might acquire each O-AGC from uncommon ancestors. We also indicated that OAg-knockout bacteria were highly adhesive in vitro, except for the EPEC001 wzy derivative, whose adherent capability was less than that of its wild-type strain, providing direct evidence that OAg plays a key role in EPEC pathogenesis. Together, we identified two EPEC O serotypes in silico and experimentally, and we also studied the adherent capabilities of their OAgs, which highlighted the fundamental and pathogenic role of OAg in EPEC.

• Journal of Microbiology and Biotechnology
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• 제34권3호
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• pp.700-709
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• 2024

Polyhydroxybutyrate (PHB) production from lignocellulosic biomass is economically beneficial. Because lignocellulosic biomass is a mixture rich in glucose and xylose, Escherichia coli, which prefers glucose, needs to overcome glucose repression for efficient biosugar use. To avoid glucose repression, here, we overexpressed a xylose regulator (xylR) in an E. coli strain expressing bktB, phaB, and phaC from Cupriavidus necator and evaluated the effect of xylR on PHB production. XylR overexpression increased xylose consumption from 0% to 46.53% and produced 4.45-fold more PHB than the control strain without xylR in a 1% sugar mixture of glucose and xylose (1:1). When the xylR-overexpressed strain was applied to sugars from lignocellulosic biomass, cell growth and PHB production of the strain showed a 4.7-fold increase from the control strain, yielding 2.58 ± 0.02 g/l PHB and 4.43 ± 0.28 g/l dry cell weight in a 1% hydrolysate mixture. XylR overexpression increased the expression of xylose operon genes by up to 1.7-fold. Moreover, the effect of xylR was substantially different in various E. coli strains. Overall, the results showed the effect of xylR overexpression on PHB production in a non-native PHB producer and the possible application of xylR for xylose utilization in E. coli.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 춘계학술발표대회
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• pp.501-504
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• 2000

The purification of a thermostable esterase expressed in Escherichia coli was investigated using thermoprecipitation of unclarified cell homogenates followed by after applying the heat-treated lysate to phenyl-sepharose column, and elution with detergent. Heat treatment at $70^{cdot}C$ was capable of removing to E. coli proteins. Specially, the thermoprecipitation with 15% polyethylene glycol 8000 can remove host proteins and nucleic acids efficiently. Various detergents were used to recover the esterase, which was strongly bound to phenyl-sepharose resin. Triton X-100, non-ionic detergent, was found to be the most efficient of all tested detergents.

• Journal of Microbiology and Biotechnology
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• 제28권7호
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• pp.1209-1216
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• 2018

This study aimed to evaluate the antimicrobial activity of 2-phenylethynyl-butyltellurium (PEBT) in Escherichia coli and the relation to its pro-oxidant effect. For this, we carried out the disk diffusion test, minimum inhibitory concentration (MIC) assay, and survival curve analysis. We also measured the level of extracellular reactive oxygen species (ROS), activity of the antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT), and level of non-protein thiols (NPSH). PEBT at 1.28 and 0.128 mg/disk exhibited antimicrobial capability in the disk diffusion test, with an MIC value of 1.92 mg/ml, whereas PEBT at 0.96, 1.92, and 3.84 mg/ml inhibited bacterial growth after a 9-h exposure. PEBT at 3.84, 1.92, and 0.96 mg/ml increased extracellular ROS production, decreased the intracellular NPSH level, and reduced the SOD and CAT activities. Glutathione or ascorbic acid in the medium protected the bacterial cells from the antimicrobial effect of PEBT. In conclusion, PEBT exhibited antimicrobial activity against E. coli, involving the generation of ROS, oxidation of NPSH, and reduction of the antioxidant defenses in the bacterial cells.

• 한국식품과학회지
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• 제31권2호
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• pp.368-374
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• 1999

식품공정에 있어서 비가열 살균 기술로 개발되고 있는 고전압 펄스 전기장(pulsed electric fields, PEF)을 과실주스의 품질 향상에 응용하고자 pilot-scale의 고전압 펄스 전기장 살균장치를 개발하였으며, 이것을 과실 주스의 모델로 선정한 사과주스의 대표적 오염지표균 또는 품질을 저하시키는 미생물 중 Escherichia coli, Bacillus subtilis, Rhodotorula minuta에 적용시켜 세포의 생리특성을 조사하였다. 내염성, 자외선 흡수 물질, 세포염색, 세포의 회복도, 세포의 표면구조 등을 조사함으로서 PEF의 미생물 생리특성에 대한 영향을 규명하고자 하였다. E. coli, B. subtilis, R. minuta를 40 kV/cm, 84 pulse, $10{\mu}s$ pulse duration의 PEF 조건에서 처리하여 내염성을 조사한 결과 1 log cycle 정도의 생존율 감소를 나타내었다. PEF 처리에 의한 세포 내부 성분의 유출 여부를 살펴본 자외선 흡수물질 측정결과 260 nm, 280 nm에서 모두 PEF 처리 세포가 무처리구에 비하여 현저하게 증가된 흡수물질 농도를 나타내었다. PEF 처리된 R. minuta를 세포 염색하여 관찰한 결과 세포막이 터져서 개열됨에 따라 염색시약이 세포 내, 외부로 고루 염색되어 세포 본래의 형태를 관찰할 수가 없었다. PEF 처리후 세포의 회복도는 무처리구에 비하여 E. coli와 B. subtilis가 약 5시간, R. minuta가 약 8시간 정도 지연된 lag time을 보였다. 그리고 PEF 처리후의 E. coli, B. subtilis, R. minuta는 정상적인 세포와는 달리 세포막이 상당한 손상을 입어 허물어진 상태를 관찰할 수 있었다.

• Asian-Australasian Journal of Animal Sciences
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• 제23권10호
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• pp.1268-1275
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• 2010

Post-weaning diarrhoea (PWD) and oedema disease (ED) caused by E. coli F18 always result in economic losses to pig producers, and no effective methods of controlling PWD and ED are presently available. FUT1 has been identified as a candidate gene controlling the expression of E. coli F18 receptor. This study examined the correlation between F18ab and F18ac adhesion phenotypes and the polymorphism at position M307 of the FUT1 gene in three pig breeds (231 Large White, 107 Landrace and 109 Songliao Black). The results showed: i) Both the susceptible genotypes (GG and GA) and the adhesion phenotypes (adhesive or weekly adhesive) were dominant in all three breeds with frequencies over 95%. ii) Three adhesion patterns of the two F18 variants F18ab and F18ac, i.e., ($ab^+$, $ac^+$), ($ab^+$, $ac^-$) and ($ab^-$, $ac^-$), were found in all three breeds, and there was no significant difference in the distribution of adhesion phenotypes of the two variants (separately or jointly) among the three breeds (p>0.05). iii) The FUT1 M307 genotypes were completely associated with the F18ab adhesion phenotypes and very strongly associated with the F18ac adhesion phenotypes. All individuals of genotype AA were non-adhesive to both F18ab and F18ac. All individuals of genotype GG or GA were adhesive to F18ab, whereas 11% of them were non-adhesive to F18ac. These results suggest that the polymorphism at FUT1 M307 can be used for marker-assisted selection of PWD and ED resistant pigs.