• 한국식품과학회지
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• 제43권5호
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• pp.611-617
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• 2011

본 연구는 열처리 방법에 의한 국내산 새싹채소 종자 3종(무, 배추, 다채)과 수입산 2종(무, 적양배추)의 발아율과 총 호기성세균 및 병원성 미생물의 저감화에 대해 실시하였다. 열처리는 40, 50, 60, 70$^{\circ}C$의 온도조건에서 각각 15, 30분 동안 실시하였다. 발아율은 60$^{\circ}C$ 이하 조건에서 열처리 후 배양 24시간 후 45-97%로 나타났으며, 대부분 70$^{\circ}C$ 이상의 조건에서 발아율이 감소하였다. 열처리에 의한 미생물 저감화 효과는 국내산 무 종자의 경우 총 호기성세균은 70$^{\circ}C$, 30분 열처리 조건에서 1.71 log CFU/g 감소하였으며 대장균군은 60$^{\circ}C$ 이상의 조건에서 검출한계이하로 나타났다. 국내산 배추 종자는 60$^{\circ}C$ 이상의 조건에서 총 호기성 세균과 대장균군은 각각 1.23-1.34, 2.77 log CFU/g 감소하였다. 국내산 다채 종자는 70$^{\circ}C$, 15분 열처리시 0.3 log CFU/g 감소되었다. 수입산 무 종자는 60$^{\circ}C$ 이상의 조건에서 2.12-2.30 log CFU/g 감소하였다. 수입산 적양배추 종자는 40$^{\circ}C$ 이상의 조건에서 0.66-0.84 log CFU/g 감소하였으며 대장균군은 검출한계 이하의 수준으로 검출되지 않았다. 대부분의 시료에서 B. cereus는 열처리에 의한 유의적인 차이가 나타나지 않았으며, 대장균, S. aureus 및 Salmonella sp.는 검출한계 수준에서 검출되지 않았다.

• Molecular & Cellular Toxicology
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• 제2권1호
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• pp.11-14
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• 2006

In this study we attempted to develop a novel genomic method to selectively isolate target functional microbial genomes from environmental samples. For this purpose, stable isotope probing (SIP) was applied in selectively isolating organic pollutant-assimilating populations. When soil microbes were fed with $^{13}C-labeled $ biphenyl, biphenyl-utilizing cells were incorporated with the heavy carbon isotope. The heavy DNA portion was successfully separated by CsCl equilibrium density gradient. And the diversity in the heavy DNA was sufficiently reduced, being suitable for the current DNA microarray techniques to detect biphenyl-utilizing populations in the soil. In addition, we proposed a new way to get more genetic information by combining this SIP method with selective metagenomic approach. The increased selective power of these new DNA isolation methods will be expected to provide a good quality of new genetic information, which, in turn, will result in development of a variety of biomarkers that may be used in assessing ecotoxicology issues including the impacts of organic hazards, and antibiotic-resistant pathogens on human and ecological systems.

• Journal of Microbiology and Biotechnology
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• 제33권10호
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• pp.1257-1267
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• 2023

Although rational genetic engineering is nowadays the favored method for microbial strain improvement, building up mutant libraries based on directed evolution for improvement is still in many cases the better option. In this regard, the demand for precise and efficient screening methods for mutants with high performance has stimulated the development of biosensor-based high-throughput screening strategies. Genetically encoded biosensors provide powerful tools to couple the desired phenotype to a detectable signal, such as fluorescence and growth rate. Herein, we review recent advances in engineering several classes of biosensors and their applications in directed evolution. Furthermore, we compare and discuss the screening advantages and limitations of two-component biosensors, transcription-factor-based biosensors, and RNA-based biosensors. Engineering these biosensors has focused mainly on modifying the expression level or structure of the biosensor components to optimize the dynamic range, specificity, and detection range. Finally, the applications of biosensors in the evolution of proteins, metabolic pathways, and genome-scale metabolic networks are described. This review provides potential guidance in the design of biosensors and their applications in improving the bioproduction of microbial cell factories through directed evolution.

• Journal of Microbiology and Biotechnology
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• 제23권12호
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• pp.1717-1725
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• 2013

Traditional soybean paste from Shandong Liangshan and Tianyuan Jiangyuan commercial soybean paste were chosen for analysis and comparison of their bacterial and fungal dynamics using denaturing gel gradient electrophoresis and 16S rRNA gene clone libraries. The bacterial diversity results showed that more than 20 types of bacteria were present in traditional Shandong soybean paste during its fermentation process, whereas only six types of bacteria were present in the commercial soybean paste. The predominant bacteria in the Shandong soybean paste were most closely related to Leuconostoc spp., an uncultured bacterium, Lactococcus lactis, Bacillus licheniformis, Bacillus spp., and Citrobacter freundii. The predominant bacteria in the Tianyuan Jiangyuan soybean paste were most closely related to an uncultured bacterium, Bacillus licheniformis, and an uncultured Leuconostoc spp. The fungal diversity results showed that 10 types of fungi were present in the Shandong soybean paste during the fermentation process, with the predominant fungi being most closely related to Geotrichum spp., an uncultured fungal clone, Aspergillus oryzae, and yeast species. The predominant fungus in the commercial soybean paste was Aspergillus oryzae.

• 한국식품과학회지
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• 제31권1호
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• pp.1-6
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• 1999

Salmonella spp.의 신속한 검출을 위하여 발진모듈, 수정결정 진동측정기, 박막형태의 수정결정으로 이루어진 압전류적(piezoelectric, PZ) 항체센서 시스템을 구성하였다. 수정결정의 금전극 표면에 Salmonella 구조 항원(common structural antigen)에 대한 항체를 protein A를 사용하여 고정화하고, 항체가 고정화된 수정결정과 미생물간의 결합반응에 의한 질량증가로 나타나는 진동수의 감소량을 측정하였다. PZ 항체센서는 $35^{\circ}C$, pH 7.2의 0.1M 인산 완충용액에서 Salmonella균에 대하여 가장 높은 감응도를 나타내었다. PZ 항체센서의 반응은 Salmonella균에 대하여 매우 선택적이었고 polystyrene bead 첨가시 센서의 감응도가 크게 증가하였다. Salmonella균의 농도가 $10^5{\sim}10^6\;CFU/mL$의 범위 내에 있을 때 쌍대수좌표에서 직선구간의 검량선을 얻을 수 있었고, Salmonella 검출에 소요되는 시간은 50분이내 이었다.

• KSBB Journal
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• 제29권5호
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• pp.361-371
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• 2014

Mycoplasma is well recognized as one of the most prevalent and serious microbial contaminants of biologic manufacturing processes. Conventional methods for mycoplasma testing, direct culture method and indirect indicator cell culture method, are lengthy, costly and less sensitive to noncultivable species. In this report, we describe a new TaqMan probe-based real-time PCR method for rapid and quantitative detection of mycoplasma contamination during manufacture of biologics. Universal mycoplasma primers were used for mycoplasma PCR and mycoplasma DNA was quantified by use of a specific TaqMan probe. Specificity, sensitivity, and robustness of the real-time PCR method was validated according to the European Pharmacopoeia. The validation results met required criteria to justify its use as a replacement for the culture method. The established real-time PCR assay was successfully applied to the detection of mycoplasma from human keratinocyte and mesenchymal stem cell as well as Vero cell lines artificially infected with mycoplasma. The overall results indicated that this rapid, specific, sensitive, and robust assay can be reliably used for quantitative detection of mycoplasma contamination during manufacture of biologics.

• 한국식품위생안전성학회지
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• 제31권6호
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• pp.465-470
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• 2016

본 연구는 전처리 방법이 새싹채소 재배에 사용되는 종자 중 식중독 세균의 검출율에 영향을 미치는지에 대해 검증하기 위해 시중 유통되고 있는 새싹채소 재배용 종자 19종을 수집하여 종자를 세척 방법과 발아 방법으로 전처리한 후 E. coli, E. coli O157:H7, Salmonella spp., L. monocytogenes의 검출율을 비교하였다. 또한 인위적으로 Salmonella enterica를 접종한 알팔파 종자에 대해서 무처리, 세척, 발아 방법으로 전처리한 후 S. enterica의 검출율을 검정하였다. 시중 유통되고 있는 종자를 수집하여 분석한 결과, 세척 방법과 발아 방법 등의 전처리 방법에 따라 E. coli 검출율과 양성시료에 차이가 있었음을 확인하였다. 인위적으로 S. enterica를 접종한 알팔파 종자에 대한 검출율 비교 검정에서도 분석시료량 대비 전처리방법별로 검출율이 차이가 나는 것으로 나타났고, 선택배지 종류에 따라서도 S. enterica의 검출율이 차이가 났다. 결론적으로 새싹채소 재배용 종자의 식중독 세균 분석에 있어서 종자의 전처리 방법, 시료당 분석시료량, 선택배지 종류 등이 검출율에 영향을 미치는 것으로 나타났다.

• Journal of Biosystems Engineering
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• 제39권3호
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• pp.215-226
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• 2014

Recently, spectroscopy has emerged as a potential tool for quality evaluation of numerous food and agricultural products because it provides information regarding both spectral distribution and image features of the sample (i.e., hyperspectral imaging). Spectroscopic techniques reveal hidden information regarding the sample and do so in a non-destructive manner. This review describes the various approaches of spectroscopic modalities, especially hyperspectroscopy and vibrational spectroscopies (i.e., Raman spectroscopy and Fourier transform near infrared spectroscopy) combined with chemometrics for the non-destructive assessment of contaminations and defects in agro-food products.

• Asian-Australasian Journal of Animal Sciences
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• 제12권1호
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• pp.129-138
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• 1999

If rumen bacteria can be manipulated to utilize nutrients (i.e., ammonia and plant cell wall carbohydrates) more completely and efficiently, the need for protein supplementation can be reduced or eliminated and the digestion of fiber in forage or agricultural residue-based diets could be enhanced. However, these approaches require a complete and accurate description of the rumen community, as well as methods for the rapid and accurate detection of microbial density, diversity, phylogeny, and gene expression. Molecular ecology techniques based on small subunit (SSU) rRNA sequences, nucleic acid probes and the polymerase chain reaction (PCR) can potentially provide a complete description of the microbial ecology of the rumen of ruminant animals. The development of these molecular tools will result in greater insights into community structure and activity of gut microbial ecosystems in relation to functional interactions between different bacteria, spatial and temporal relationships between different microorganisms and between microorganisms and reed panicles. Molecular approaches based on SSU rRNA serve to evaluate the presence of specific sequences in the community and provide a link between knowledge obtained from pure cultures and the microbial populations they represent in the rumen. The successful development and application of these methods promises to provide opportunities to link distribution and identity of gastrointestinal microbes in their natural environment with their genetic potential and in situ activities. The use of approaches for assessing pupulation dynamics as well as for assessing community functionality will result in an increased understanding and a complete description of the gastrointestinal communities of production animals fed under different dietary regimes, and lead to new strategies for improving animal growth.

• 한국축산식품학회지
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• 제35권5호
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• pp.674-682
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• 2015

The objective of this study was to evaluate the risk of illness from Campylobacter spp. on ham. To identify the hazards of Campylobacter spp. on ham, the general characteristics and microbial criteria for Campylobacter spp., and campylobacteriosis outbreaks were investigated. In the exposure assessment, the prevalence of Campylobacter spp. on ham was evaluated, and the probabilistic distributions for the temperature of ham surfaces in retail markets and home refrigerators were prepared. In addition, the raw data from the Korea National Health and Nutrition Examination Survey (KNHNES) 2012 were used to estimate the consumption amount and frequency of ham. In the hazard characterization, the Beta-Poisson model for Campylobacter spp. infection was used. For risk characterization, a simulation model was developed using the collected data, and the risk of Campylobacter spp. on ham was estimated with @RISK. The Campylobacter spp. cell counts on ham samples were below the detection limit (<0.70 Log CFU/g). The daily consumption of ham was 23.93 g per person, and the consumption frequency was 11.57%. The simulated mean value of the initial contamination level of Campylobacter spp. on ham was −3.95 Log CFU/g, and the mean value of ham for probable risk per person per day was 2.20×10⁻¹². It is considered that the risk of foodborne illness for Campylobacter spp. was low. Furthermore, these results indicate that the microbial risk assessment of Campylobacter spp. in this study should be useful in providing scientific evidence to set up the criteria of Campylobacter spp..