• The Korean Journal of Ecology
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• 제18권1호
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• pp.109-120
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• 1995

As the genetically modified microorganisms (GMMs) and their recombinant plasmid DNAs could be released into natural environments, their stabilities and impacts to indigenous microorganisls have become very importhant research subjects concerning with environmental and ecological aspects. In this study, the genetically modified E. coli CU103 and its recombinant pCU103 plasmid DNA, in which pcbCD genes involving in degradation of biphenyl and 4-chlorobiphenyl were cloned, were studied for their survival and stability in several different waters established under laboratory conditions. E. coli CU103 and its host E. coli XL1-Blue survived longer in sterile distilled water (SDW) and filtered autoclaved river water (FAW) than in filtered river water (FW). A lot of extracellular DNAs were released from E. coli CU103 by lytic action of phages in FW and the released DNAs were degraded by DNase dissolved in the water. Such effects of the factors in FW on stability of the recombinant pCU103 plasmid were also observed in the results of gel electrophoresis, quantitative analysis with bisbenzimide, and transformation assay. Therefore, the recombinant plasmids of pCU103 were found to be readily liberated from the genetically modified E. coli CU103 into waters by normal metabolic processes and lysis of cells. And the plasmid DNAs were quite stable in waters, but their stabilities could be affected by physicoKDICical and biological factors in non-sterile natural waters.

• Bulletin of the Korean Chemical Society
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• 제27권10호
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• pp.1681-1684
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• 2006

• 한국토양비료학회지
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• 제46권6호
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• pp.647-651
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• 2013

Animal manure compost is a commonly used fertilizer in organic vegetable and fruit production in Korea. However, livestock manure compost produced from animal feces can contain a lot of the non-pathogenic and pathogenic bacteria. Of particular concern are bacteria causing human food-borne illness such as Escherichia coli O157:H7 and Listeria monocytogenes. The objective of this study was to investigate effect of temperature on survival of E. coli O157:H7 and L. monocytogenes in livestock manure compost. Commercial livestock manure compost (manure 60%, sawdust 40%) was inoculated with E. coli O157:H7 and L. monocytogenes. Compost was incubated at four different temperatures (10, 25, 35, and $55^{\circ}C$) for 20 weeks. Samples were taken every week during incubation depending on the given conditions. E. coli O157:H7 persisted for up to 1 day in livestock manure compost at $55^{\circ}C$, over 140 days at $10^{\circ}C$, 140 days at $25^{\circ}C$, and 120 days at $35^{\circ}C$, respectively. L. monocytogenes persisted for up to 1 day in livestock manure compost at $55^{\circ}C$ and 140 days at $10^{\circ}C$, 70 days at $25^{\circ}C$, and 40 days at $35^{\circ}C$, respectively. The results indicated that E. coli O157:H7 and L. monocytogenes persisted longer under low temperature condition. E. coli O157:H7 survived longer than L. monocytogenes at three different temperatures (10, 25, and $35^{\circ}C$). The results are being used to develop guidelines on the management of manure to reduce the risks of E. coli O157:H7 and L. monocytogenes transmission to foods produced in the presence of animal waste.

• 한국생명과학회:학술대회논문집
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• 한국생명과학회 2001년도 제34회 학술심포지움
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• pp.3-8
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• 2001

Isopentenyl diphosphate (IPP) is the common, five-carbon building block in the biosynthesis of all carotenoids, IPP in Escherichia coli is synthesized through the non-mevalonate pathway. The first reaction of IPP biosynthesis in E. coli is the formation of 1-deoxy-D-xylulose-5-phosphate (DXP), catalyzed by DXP synthase and encoded by dxs. The second reaction in the pathway is the reduction of DXP to 2-C-methyl-D-erythritol-4-phosphate, catalyzed by DXP reductoisomerase and encoded by dxr. To determine if one or more of the reactions in the non-mevalonate pathway controlled flux to IPP, dxs and dxr were placed on several expression vectors under the control of three different promoters and transformed into three E. coli strains (DH5(, XL1-Blue, and JM101) that had been engineered to produce lycopene. Lycopene production was improved significantly in strains transformed with the dxs expression vectors. When the dxs gene was expressed from the arabinose-inducible araBAD promoter (PBAD) on a medium-copy plasmid, lycopene production was 2-fold higher than when dxs was expressed from the IPTG-inducible trc and lac promoters (Ptrc and Plac, respectively) on medium-copy and high-copy plasmids, Given the low final densities of cells expressing dxs from IPTG-inducible promoters, the low lycopene production was probably due to the metabolic burden of plasmid maintenance and an excessive drain of central metabolic intermediates. At arabinose concentrations between 0 and 1.33 mM, cells expressing both dxs and dxr from PBAD on a medium-copy plasmid produced 1.4 - 2.0 times more lycopene than cells expressing dxs only. However, at higher arabinose concentrations lycopene production in cells expressing both dxs and dxr was lower than in cells expressing dxs only. A comparison of the three E. coli strains transformed with the arabinose-inducible dxs on a medium-copy plamid revealed that lycopene production was highest in XL1-Blue.

• Fisheries and Aquatic Sciences
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• 제22권11호
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• pp.26.1-26.7
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• 2019

Background: The monitoring of pathogens of fishery auction markets is important to obtain safe fishery products regarding hygiene and sanitation. In this study, aerobic, coliform, Escherichia coli, and Vibrio cholerae were monitored in the fishery products and environmental samples obtained from fishery auction markets. Methods: The fishery products (flounder, octopus, skate, rock cod, sea bass, snail, monkfish, flatfish, comb pen shell, corb shell, conger eel, hairtail, croaker, and pilchard) were placed in filter bags, and the environmental samples (samples from the water tanks at the fishery auction markets, seawater from the fishery distribution vehicles, ice from wooden or plastic boxes, and surface samples from wooden and plastic boxes used for fish storage) were collected. Aerobic bacteria, E. coli, and coliform in the samples were enumerated on aerobic count plates and E. coli/coliform count plates, respectively. For V. cholerae O1 and V. cholerae non-O1 quantification, most probable number (MPN)-PCR analysis was performed. Results: Aerobic and coliform bacteria were detected in most samples, but E. coli was not detected. Wooden boxes were contaminated with high levels of aerobic and coliform bacteria in all seasons (spring, summer, and fall). During fall, V. cholerae non-O1 were detected in snails, hairtails, croakers, flatfishes, pilchards, plastic boxes, and water samples. Conclusions: These results indicate an increased prevalence of V. cholerae contamination in fishery products in fall, including food contact samples, which can be vehicles for cross-contamination.

• 한국식품영양학회지
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• 제36권1호
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• pp.17-24
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• 2023

To apply UV-C as a non-heating sterilization method to increase the microbiological safety of fresh seedless watermelon products, reductions in E. coli and quality changes by treatment dose (0, 2, 4, 8, 14, 20 kJ/m²) were investigated. The pH, sugar content, and hardness of watermelon inoculated with E. coli were not significantly different according to the UV-C treatment dose, but the polyphenol content was significantly decreased compared to the controls (425.4 GAE ㎍/g F.W.). When treated with 2 and 4 kJ/m², the lycopene content was 31.6 and 30.9 ㎍/g F.W., respectively, which was increased compared to the controls (28.5 ㎍/g F.W.). The arginine and citrulline content was also significantly increased compared to the controls. The number of E. coli was significantly decreased compared to the controls following UV-C treatment. Considering the degree of E. coli reduction, lycopene content, arginine content, citrulline content, and UV-C irradiation time, subsequent experiments were conducted by selecting a UV-C treatment dose of 2 kJ/m². The results of confirming the degree of reduction in the number of E. coli colonies by a single treatment and combined treatment with UV-C 2 kJ/m² and 70% ethanol showed that the combined treatment was most effective as colonies were decreased by 2.3 log CFU/g compared to the controls. Therefore, it is judged that UV-C 2 kJ/m² radiation and combined treatment with 70% ethanol could be applied as a non-heating sterilization method for fresh watermelon slices.

• 한국식품위생안전성학회지
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• 제38권1호
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• pp.31-36
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• 2023

본 연구는 집단급식소에서 제공되는 빈도수가 높은 비가열 및 가열조리 엽경채류에 사용되는 차아염소산나트륨 수에 대하여 미생물적 안전성을 평가하고자 수행되었다. 비병원성 대장균과 장출혈성 대장균의 칵테일(E. coli O157:H7)을 엽경채류(초기 균수 7-8 log CFU/g)에 인위적으로 오염시킨 후 차아염소산나트륨을 5분간 침지 후 흐르는 물에 3번 씻어서 생균수를 측정하였다. 실험 결과 초기 오염물질에 비해 살균효과가 1-2 log CFU/g 저감화하여 대조군에 대해 유의적인 차이가 있었다(P<0.05). 잎채소의 특성에 따라 약간의 차이가 있었는데 표면적이 클수록, 덜 거칠고 잎이 부드러울수록 살균효과가 높았다. 200 mg/kg으로 처리하였을 때 100 mg/kg에 비해 0.1-0.3 log CFU/g만큼 효과가 더 감소하였으나 농도 증가에 따른 유의적 차이는 없었다(P>0.05). 그러므로 학교급식위생관리지침에서 제시한 기준 이상으로 차아염소산나트륨 농도를 높이는 것은 불필요하다고 판단된다. 그러나 잎채소는 일반적으로 미생물의 초기 오염도가 높기 때문에 차아염소산나트륨 처리만으로는 안전한 수준의 저감을 달성하기 어려워 생물학적 위험이 잔존한다. 따라서 여름철에 가열하지 않은 잎채소의 대체 조리방법을 개발하는 것이 안전성에 보다 효과적인 것으로 판단된다.

• 한국식품영양과학회지
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• 제21권1호
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• pp.76-83
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• 1992

장독성대장균은 영유아설사 및 여행자설사의 주요한 원인균의 하나로서 이들이 생산하는 내열성장독소는 설사 유발 원인물질로 이들 생산균주는 비병원성 대장균과 비교하여 검정할 수 있다. 본 실험에서는 이러한 내열성장독소를 순수정제하였다. 저자들이 보고한 바 있는 형질전환균주 eKT-53을 사용하여 10L의 succinate salts 배지에 배양한 배양액을 30,000g로 원심분리하여 조 ST 용액을 사용하여 몇 단계의 정제과정을 통하여 정제하였다. 즉 Amberlite XAD-2 chromatography, DEAE-Sephacel anion exchange chromatography, Bio-Gel P-6 get filtration, Polyacrylamide slab gel 전기영동을 통하여 정제하고 장독소의 정제도 및 장독소의 생물학적 최소단위량을 조사하였다. 최종적으로 정제되어진 ST는 조 ST 용액에 비해 113배 더 정제되었고 회수율은 11%였다. 또한 생물학적 최소단위량이 2.8ng이였고 SDS-polyacrylamide disc gel상에서 단일 band를 나타내어 거의 순수정제된 것으로 보이며 면역을 위한 항원으로 이용할 수 있을 것으로 생각된다.

• Journal of Microbiology and Biotechnology
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• 제25권8호
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• pp.1339-1348
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• 2015

Until now, considerable effort has been made to engineer novel nitrogen-fixing organisms through the transfer of nif genes from various diazotrophs to non-nitrogen fixers; however, regulatory coupling of the heterologous nif genes with the regulatory system of the new host is still not well understood. In this work, a 49 kb nitrogen fixation island from P. stutzeri A1501 was transferred into E. coli using a novel and efficient transformation strategy, and a series of recombinant nitrogen-fixing E. coli strains were obtained. We found that the nitrogenase activity of the recombinant E. coli strain EN-01, similar to the parent strain P. stutzeri A1501, was dependent on external ammonia concentration, oxygen tension, and temperature. We further found that there existed a regulatory coupling between the E. coli general nitrogen regulatory system and the heterologous P. stutzeri nif island in the recombinant E. coli strain. We also provided evidence that the E. coli general nitrogen regulator GlnG protein was involved in the activation of the nif-specific regulator NifA via a direct interaction with the NifA promoter. To the best of our knowledge, this work plays a groundbreaking role in increasing understanding of the regulatory coupling of the heterologous nitrogen fixation system with the regulatory system of the recipient host. Furthermore, it will shed light on the structure and functional integrity of the nif island and will be useful for the construction of novel and more robust nitrogen-fixing organisms through biosynthetic engineering.

• 한국미생물·생명공학회지
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• 제7권3호
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• pp.165-173
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• 1979

자연선조사한 대장균B주의 세포분열회복활성분을 구명코저 자외선내성균인 대장균 B/r 주의 초음파추출액으로부터 활성성분分을 분리한 결과 $\beta$-NAD가 관여함이 발표되었다. 본고에서는$\beta$-NAD 이외 Magnesium이 활성물질의 안정화가 중요한 역할을 나타냄을 구명하였으며 10~30%의 서당밀 도구배원심분리에 의해 2 개의 새로운 활성부분이 있음을 확인하였다. 2 개의 활성물질 가운데 하나는 원심관의 최하부에 위치하였으며 또 다른 하나는 상부의 분자량 45,000 부위에서 회수되었다. 하부에 위치한 활성획분은 Mg++이 그 활성에 무관하였으나 상부의 저분자 활성부분은 $Mg^{++}$을 첨가하지 않으면 회수가 불가능하였다. 저분자활성부분은 pronase에 대해 감수성이었으며 DNA-ligase 는 아님이 추정되었다. 초원심분리과정에 $N_2$ gas를 처리할 경우 aeration에 비해 약 2 배의 활성이 나타났다. $Mg^{++}$은 $\beta$-NAD에 또하나의 회복활성 및 필수적인자로 요구된다고 생각된다.