• 제목/요약/키워드: E. coli system

검색결과 739건 처리시간 0.022초

우유내의 LP system의 생리기능 및 항균성에 관한여구 1. Escherichia coli O157:H7에 대한 항균효과 (Studies on the Biological Function and Antibacterial Effect of Lactoperoxidase System in Milk)

  • 정충일;남은숙;김대원;전형일
    • 한국식품위생안전성학회지
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    • 제13권1호
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    • pp.1-5
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    • 1998
  • 본 연구는 E. coli O157:H7에 대한 LP system(lactoperoxidae/thiocyanate/hydrogen peroxide)의 항균효과를 측정하기 위해 수행되었다. 초기 접종수준($10^{2},\;10^{4},\;10^{7}cfu/ml$), LP의 농도 (10ppm, 20ppm, 30ppm), 배지종류 (TSB, UHT milk, raw milk), 배양온도 ($5^{\circ}C,\;10^{\circ}C,\;15^{\circ}C$) 등에 따라 E. coli O157:H7에 대한 항균효과를 측정, 비교한 결과, 초기 접종수준을 $10^2/ml$으로 하였을 때와 LP의 농도를 10ppm 및 $5^{\circ}C$ 배양에서 항균력이 가장 높게 나타났다.

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Cellulomonas biazotea cellobiase gene의 대장균에의 형질발현 (Exprission of cellulomonas biazotea cellobiase gene in E. coli)

  • 박영길;연창석;최영길
    • 미생물학회지
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    • 제26권1호
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    • pp.6-12
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    • 1988
  • Cellobiase ($\beta$-glucosidase) is an enzyme of the cellulase system in cellulolytic microor-ganisms. The chromosomal DNA fragment which include cellobiase gene of Cellulomonas biazotea was cloned in Eschericia coli via plasmid pBR 322 vector. Restriction enzyme Sal I was used to obtain adequate size of fragments from C. biazotea. chromosomal DNA. The transformant of E. coli HB101 with recombinant plasmid pBG101 showed cellobiase activity, which is not ordinary in E. coli HB101. The enzyme activity of the transformant was as of 20% lower than that of C. biazotea.

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Immunosensor for Detection of Escherichia coli O157:H7 Using Imaging Ellipsometry

  • Bae Young-Min;Park Kwang-Won;Oh Byung-Keun;Choi Jeong-Woo
    • Journal of Microbiology and Biotechnology
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    • 제16권8호
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    • pp.1169-1173
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    • 2006
  • Imaging ellipsometry (IE) for detection of binding of Escherichia coli O157:H7 (E. coli O157:H7) to an immunosensor is reported. A protein G layer, chemically bound to a self-assembled layer of 11-mercaptoundecanoic acid (11-MUA), was adopted for immobilization of monoclonal antibody against E. coli O157:H7 (Mab). The immobilization of antibody was investigated using surface plasmon resonance. To fabricate antibody spots on a gold surface, protein G solution was spotted onto the gold surface modified with an 11-MUA layer, followed by immobilizing Mab on the protein G spot. Ellipsometric images of the protein G spot, the Mab spot, and Mab spots with binding of E. coli O157:H7 in various concentrations were acquired using the IE system. The change of mean optical intensity of the Mab spots in the ellipsometric images indicated that the lowest detection limit was $10^3$CFU/ml for E. coli O157:H7. Thus, IE can be applied to an immunosensor for detection of E. coli O157:H7 as a detection method with the advantages of allowing label-free detection, high sensitivity, and operational simplicity.

Comparative Analysis of Tagatose Productivity of Immobilized L-Arabinose Isomerase Expressed in Escherichia coli and Bacillus subtilis

  • Cheon, Ji-Na;Kim, Seong-Bo;Park, Seong-Won;Han, Jong-Kwon;Kim, Pil
    • Food Science and Biotechnology
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    • 제17권3호
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    • pp.655-658
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    • 2008
  • Although arabinose isomerase (E.C. 5.3.1.4), a commercial enzyme for edible tagatose bioconversion, can be expressed in an Escherichia coli system, this expression system might leave noxious by-products in food. To develop an eligible tagatose bioconversion with food-safe system, we compared the tagatose production activity of immobilized arabinose isomerase expressed in Bacillus subtilis (a host generally recognized as safe) with that of the enzyme expressed in E. coli. A 48% increase in tagatose production (4.3 g tagatose/L at $69.4\;mg/L{\cdot}hr$) was found using the B. subtilis-expressed immobilized enzyme system, compared to the E. coli-expressed enzyme system (2.9 g tagatose/L). The increased productivity with safety of the B. subtilis-expressed arabinose isomerase suggests that it is a more eligible candidate for commercial tagatose production.

Microwave 조사에 의한 Salmonella enteritidis, Salmonella typhimurium 과 E. coli의 불활성에 관한 연구 (Thermal Inactivation of Salmonella enteritidis, Salmonella typhimurium and E. coli O111 in Liquid Cultures During Microwave Radiation)

  • 이조윤;이강욱;배형철;김종우
    • 한국축산식품학회지
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    • 제18권3호
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    • pp.269-275
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    • 1998
  • The purpose of this study was to determine the thermal inactivation of Salmonella enteritidis, Salmonella typhimurium and E. coli O111 in liquid cultures treated with microwave energy. Furthermore, this study was to introduce new methodologies for studying nonthermal microwave effects on microorganisms, using controlled microwave energy and specially designed apparatuses. For the automatic temperature control during microwave heating, the real time data acquisition and computation system is designed with BASIC routine. The automatic temperature control system used in the experiments perform relatively stable control at the experiment temperature of 45, 50, 55 60$^{\circ}C$ and 65$^{\circ}C$ for 30 minutes. The effects of microwave heating on liquid cultures was compared with that of conventional heating, still reduces effectively the number of pathogenic bacteria in liquid cultures. While no particular differences between microwave heating and conventional heating was observed in the activation of E. coli at 45$^{\circ}C$ test, the activation of Sal. enteritidis and Sal. typhimurium was slightly reduced during the microwave treatments.

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Growth Inhibition of Escherichia coli during Heterologous Expression of Bacillus subtilis Glutamyl-tRNA Synthetase that Catalyzes the Formation of Mischarged Glutamyl-$tRNA_{l}$$^{Gln}$

  • Baick, Ji-Won;Yoon, Jang-Ho;Suk Namgoong;Dieter Soll;Kim, Sung-Il;Eom, Soo-Hyun;Hong, Kwang-Won
    • Journal of Microbiology
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    • 제42권2호
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    • pp.111-116
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    • 2004
  • It is known that Bacillus subtilis glutamyl-tRNA synthetase (GluRS) mischarges E. coli $tRNA_{1}$$^{Gln}$ with glutamate in vitro. It has also been established that the expression of B. subtilis GluRS in Escherichia coli results in the death of the host cell. To ascertain whether E. coli growth inhibition caused by B. subtilis GluRS synthesis is a consequence of Glu-$tRNA_{1}$$^{Gln}$ formation, we constructed an in vivo test system, in which B. subtilis GluRS gene expression is controlled by IPTG. Such a system permits the investigation of factors affecting E. coli growth. Expression of E. coli glutaminyl-tRNA synthetase (GlnRS) also amelio-rated growth inhibition, presumably by competitively preventing $tRNA_{1}$$^{Gln}$ misacylation. However, when amounts of up to 10 mM L-glutamine, the cognate amino acid for acylation of $tRNA_{1}$$^{Gln}$, were added to the growth medium, cell growth was unaffected. Overexpression of the B. subtilis gatCAB gene encoding Glu-$tRNA^{Gln}$ amidotransferase (Glu-AdT) rescued cells from toxic effects caused by the formation of the mis-charging GluRS. This result indicates that B. subtilis Glu-AdT recognizes the mischarged E. coli Glu-$tRNA_{1}$$^{Gln}$, and converts it to the cognate Gln-$tRNA_{1}$$^{Gln}$ species. B. subtilis GluRS-dependent Glu-$tRNA_{1}$$^{Gln}$ formation may cause growth inhibition in the transformed E. coli strain, possibly due to abnormal protein synthesis.

정수처리공정에 따른 일반세균과 대장균군의 제거에 관한 연구 (A Study on the Removal Effect of Bacteria and E. Coli. by Water Treatment Processes using Activated Carbon and Membrane)

  • 조태석;김영규;정문호
    • 한국환경보건학회지
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    • 제23권2호
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    • pp.24-27
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    • 1997
  • This study has been designed to check the removal effect of contaminated water by various water treatmemt processes using sediment filter, activated carbon, reverse osmosis membrane, ultra vilolet sterilizer and ultra filtration and then to analyze the change of pH, the concentration of chlorides, bacteria and E. coli. after 24 hours. pH has increased as much as 0.15-0.32 by activated carbon but decreased sharply by reverse osmosis treatment after 24 hours. The removal effect of chloride was low by activated carbon and ultra filter but high in reverse osmosis. The removal effect of bacteria and E. coli was low by activated carbon and membrane filter system using activated carbon. Ultra filtration process was effective for purify agricultural water containg bacteria and E.coli.

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Engineering of a Microbial Cell Factory for the Extracellular Production of Catalytically Active Phospholipase A2 of Streptomyces violaceoruber

  • Lee, Hyun-Jae;Cho, Ara;Hwang, Yeji;Park, Jin-Byung;Kim, Sun-Ki
    • Journal of Microbiology and Biotechnology
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    • 제30권8호
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    • pp.1244-1251
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    • 2020
  • Phospholipase A2 (PLA2) from Streptomyces violaceoruber is a lipolytic enzyme used in a wide range of industrial applications including production of lysolecithins and enzymatic degumming of edible oils. We have therefore investigated expression and secretion of PLA2 in two workhorse microbes, Pichia pastoris and Escherichia coli. The PLA2 was produced to an activity of 0.517 ± 0.012 U/ml in the culture broth of the recombinant P. pastoris. On the other hand, recombinant E. coli BL21 star (DE3), overexpressing the authentic PLA2 (P-PLA2), showed activity of 17.0 ± 1.3 U/ml in the intracellular fraction and 21.7 ± 0.7 U/ml in the culture broth. The extracellular PLA2 activity obtained with the recombinant E. coli system was 3.2-fold higher than the corresponding value reached in a previous study, which employed recombinant E. coli BL21 (DE3) overexpressing codon-optimized PLA2. Finally, we observed that the extracellular PLA2 from the recombinant E. coli P-PLA2 culture was able to hydrolyze 31.1 g/l of crude soybean lecithin, an industrial substrate, to a conversion yield of approximately 95%. The newly developed E. coli-based PLA2 expression system led to extracellular production of PLA2 to a productivity of 678 U/l·h, corresponding to 157-fold higher than that obtained with the P. pastoris-based system. This study will contribute to the extracellular production of a catalytically active PLA2.

MOLECULAR BREEDING OF GLUTATHIONE PRODUCING BACTERIAL STRAINS

  • 남용석
    • 한국미생물학회:학술대회논문집
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    • 한국미생물학회 1991년도 춘계학술발표대회 논문집
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    • pp.237-242
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    • 1991
  • In order to increase the production of glutathione by maximizing the expression of recombinant gsh plasmids, two genes responsible for the biosynthesis of glutathione were cloned. A gshI gene was cloned onto pBR322 plasmid as 3.6Kb PstI DNA fragment from E. coli K-12 chromosomal DNA. Also gshII gene was cloned onto pUC13 plasmid as 2.2Kb PstI-BamHI DNA fragment. In order to improve the glutathione producing activity more efficiently, various recombinant plasmids containing tandem repeated gshI genes or both genes in various copy number onto the same vector were constructed. E. coli cells harboring pGH501 plasmid (pUC8-gshI$\cdot$I$\cdot$II) showed the highest glutathione synthesizing activity. The conditions for glutathione production with an ATP-generating system such as acetate kinase reaction of E. coli cells or glycolytic pathway of yeast cells were examined using the E. coli cells harboring the pGH501 plasmid. When the acetate kinase reaction of E. coli cells was used as an ATP generating system, 20mM of L-csteine was converted into glutathione with a yield of $100\%$.

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Construction of Methanol-Sensing Escherichia coli by the Introduction of a Paracoccus denitrificans MxaY-Based Chimeric Two-Component System

  • Ganesh, Irisappan;Vidhya, Selvamani;Eom, Gyeong Tae;Hong, Soon Ho
    • Journal of Microbiology and Biotechnology
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    • 제27권6호
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    • pp.1106-1111
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    • 2017
  • Escherichia coli was engineered to sense methanol by employing a chimeric two-component system (TCS) strategy. A chimeric MxaY/EnvZ (MxaYZ) TCS was constructed by fusing the Paracoccus denitrificans MxaY with the E. coli EnvZ. Real-time quantitative PCR analysis and GFP-based fluorescence analysis showed maximum transcription of ompC and the fluorescence at 0.01% of methanol, respectively. These results suggested that E. coli was successfully engineered to sense methanol by the introduction of chimeric MxaYZ. By using this strategy, various chimeric TCS-based bacterial biosensors can be constructed and used for the development of biochemical-producing recombinant microorganisms.