• 제목/요약/키워드: Non-E. coli

검색결과 367건 처리시간 0.032초

Expression of Cyclodextrinase Gene from Paenibacillus sp. A11 in Escherichia coli and Characterization of the Purified Cyclodextrinase

  • Kaulpiboon, Jarunee;Pongsawasdi, Piamsook
    • BMB Reports
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    • 제37권4호
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    • pp.408-415
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    • 2004
  • The expression of the Paenibacillus sp. A11 cyclodextrinase (CDase) gene using the pUC 18 vector in Escherichia coli JM 109 resulted in the formation of an insoluble CDase protein in the cell debris in addition to a soluble CDase protein in the cytoplasm. Unlike the expression in Paenibacillus sp. A11, CDase was primarily observed in cytoplasm. However, by adding 0.5 M sorbitol as an osmolyte, the formation of insoluble CDase was prevented while a three-fold increase in cytoplasmic CDase activity was achieved after a 24 h-induction. The recombinant CDase protein was purified to approximately 14-fold with a 31% recovery to a specific activity of 141 units/mg protein by 40-60% ammonium sulfate precipitation, DEAE-Toyopearl 650 M, and Phenyl Sepharose CL-4B chromatography. It was homogeneous by non-denaturing and SDS-PAGE. The enzyme was a single polypeptide with a molecular weight of 80 kDa, as determined by gel filtration and SDS-PAGE. It showed the highest activity at pH 7.0 and $40^{\circ}C$. The catalytic efficiency ($k_{cat}/K_m$) values for $\alpha$-, $\beta$-, and $\gamma$-CD were $3.0{\times}10^5$, $8.8{\times}10^5$, and $5.5{\times}10^5\;M^{-1}\;min^{-1}$, respectively. The enzyme hydrolyzed CDs and linear maltooligosaccharides to yield maltose and glucose with less amounts of maltotriose and maltotetraose. The rates of hydrolysis for polysaccharides, soluble starch, and pullulan were very low. The cloned CDase was strongly inactivated by N-bromosuccinimide and diethylpyrocarbonate, but activated by dithiothreitol. A comparison of the biochemical properties of the CDases from Paenibacillus sp. A11 and E. coli transformant (pJK 555) indicates that they were almost identical.

Syntheses and Characterizations of Serine and Threonine Capped Water-Dispersible ZnS:Mn Nanocrystals and Comparison Study of Toxicity Effects on the growth of E. coli by the Methionine, Serine, Threonine, and Valine Capped ZnS:Mn Nanocrystals

  • Lim, Eun-Ju;Park, Sang-Hyun;Byun, Jong-Hoe;Hwang, Cheong-Soo
    • Bulletin of the Korean Chemical Society
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    • 제33권5호
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    • pp.1741-1747
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    • 2012
  • Water-dispersible ZnS:Mn nanocrystals were synthesized by capping the surface of the nanocrystals with conventional aminoacids ligands: serine and threonine. The aminoacids capped ZnS:Mn nanocrystal powders were characterized by XRD, HR-TEM, EDXS, ICP-AES and FT-IR spectroscopy. The optical properties were also measured by UV/Vis and solution photoluminescence (PL) spectroscopies in aqueous solvents. The solution PL spectra showed broad emission peaks around 600 nm with PL efficiencies of 9.7% (ZnS:Mn-Ser) and 15.4% (ZnS:Mn-Thr) respectively. The measured particle sizes for the aminoacid capped ZnS:Mn nanocrystals by HR-TEM images were about 3.0-4.0 nm, which were also supported by Debye-Scherrer calculations. In addition, cytotoxic effects of four aminoacids capped ZnS:Mn nanocrsystals over the growth of wild type E. coli were investigated. Although toxicity in the form of growth inhibition was observed with all the aminoacids capped ZnS:Mn nanocrystals at higher dose (1 mg/mL), ZnS:Mn-Met and ZnS:Mn-Thr appeared non-toxic at doses less than 100 ${\mu}g$/mL. Low biological toxicities were seen at doses less than 10 ${\mu}g$/ mL for all nanocrystals.

세균 배양액과 반응한 Lactobacillus plantarum의 생장속도 및 항균 활성에 대한 연구 (Growth Rate and Antimicrobial Activity of Lactobacillus plantarum Reacting with Bacterial Culture Supernatant)

  • 하은미
    • 한국미생물·생명공학회지
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    • 제43권3호
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    • pp.187-194
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    • 2015
  • 인간의 위장관은 매우 복잡한 장내균총으로 이루어져 있으며, 장내균총은 인간의 생리 기능에 있어 매우 중요한 역할을 담당하고 있다. 특히 유아기의 장내균총의 불균형은 면역연관 질병을 일으키는 요인이며 치료 목적으로 probiotics가 유용하게 활용되고 있다. Probiotics의 대표 균종은 L. plantarum으로서 영유아기에 병원균보다 가장 먼저 정착해야 면역 질환을 예방할 수 있다. 그러나 아직 Lactobacillus가 장의 우점종으로 정착하기 위한 환경적 요인과 항생물질분비 조절에 대한 이해는 부족한 실정이다.

내산성, 내답즙성이 높은 미생물을 이용한 생균제 개발

  • 김소영;정해영;조철희;박근형;손석민;이기영;이건순;김홍;채희정
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2003년도 생물공학의 동향(XII)
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    • pp.180-184
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    • 2003
  • 선별된 7종의 균주는 넓은 범위의 산과 담즙산에 대해 기존에 보고된 결과에 비하여 높은 내성을 나타냈고, 장내세균인 E. coli에 대해서도 우수한 항균력을 나타냈고, 항생제 tetracycline, nisin, kanamycin, ampicillin. streptomycin에 대해서도 높은 내성을 보였다. 또한 7종의 균주는 어느 정도의 산을 생성하고 있었고, 네 가지 효소 amylase, protease, lipase, cellulase에 대해서도 높은 활성을 나타냈다. 균주 특성실험 결과, 7종의 균주는 그람음성균으로 모두 구형, 비운동성이고 균체의 색은 노란색이거나 흰색을 띠고 있고, 다양한 유기물에 대하여 분해력을 지니고 있었다.

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Halomonas subglaciescola DH-1의 생장에 미치는 염화나트륨의 영향 (Effect of NaCl on Halomonas subglaciescola DH-1 Incapable of Growing at Non-Salinity)

  • 나병관;유영선;박두현
    • 한국미생물·생명공학회지
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    • 제35권4호
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    • pp.298-303
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    • 2007
  • 호염성세균 H. subglaciescola DH-1은 염화나트륨이 없거나 0.8 M 이하로 존재하는 환경에서 생장하지 못한다. 이 호혐성세균은 2.0 M의 염화나트륨이 존재하는 조건에서는 최적온도($30^{\circ}C$)보다 높은 $40^{\circ}C$에서 생장이 가능하였으나, 0.8 M의 염화나트륨이 존재하는 조건에서는 생장이 크게 저하되었다. 세포추출물을 염화나트륨이 존재하는 조건에서 $50^{\circ}C$로 1시간 동안 열처리하였을 때 세포내 효소의 활성이 유지되었으나, 염화나트륨이 없는 조건에서 열처리하였을 때 효소의 활성은 유지되지 않았다. 반면, 대장균의 세포추출물의 효소활성은 1.0 M이상의 염화나트륨이 존재할 때 온도 또는 pH와 관계없이 측정되지 않았다. H. subglaciescola DH-1은 pH $7{\sim}10$의 범위에서 생장하였고, 생장을 위한 최적 pH는 8이었다. 이러한 생리적인 특성으로부터 염화나트륨은 H. subglaciescola DH-1의 물질대사를 위한 필수적인 무기영양소라는 사실을 유추할 수 있다.

통계적 분석을 이용한 간이급수시설의 오염원에 관한 연구 (A Study on the Pollution Sources of Simple water Supply Piped System using Statistical Analysis)

  • 이홍근;김현용;백도현;김지영;이태호
    • 환경위생공학
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    • 제14권2호
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    • pp.56-67
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    • 1999
  • This study was performed to suggest the basic data and plans for the establishment of safe water supply plans in simple water supply piped system in the rural areas. In 4 different places, 24 points of water sources 36 points of taps from water sources were sampled. Of the whole 60 points, 55 points were ground water and 5 points were surface water. 14 items were measured for the analysis of water quality on each samples. The measured items were analyzed again by statistical method ; cluster analysis and principle components analysis. The results of this study are as followed. 1) In water quality analysis on water sources, 4 items, bacteria, E.coli, NH3-N and Fe exceed the standard. Of 24 points, 20 points(83%) on bacteria, 1 point(4%) on NH3-N and Fe exceed the standard. 2) In water quality analysis on near and remote taps, 4 items, bacteria, E.coli, NH3-N and Fe , exceed the standard. Of 36 points, 20 points (81%) on bactria, 1 pint(3%) on NH3-N and Fe exceed the standard. 3)Cluster analysis on water quality shows the differences by the kinds of water sources, geographical characteristics and distance from water sources. 4) Principle components analysis on ground water shows that Factor 1 and Factor 3 are natural fluctuation by the content of soil. Also, Factor 2 and Factor 4 are penetration of pollutants to underground. Therefore, it is needed to take deeper ground water in order to prevent from pollution in the areas which have ground water as water source . 5) Principle components analysis on surface water shows that Factor 1 is penetration of vacteria from surface to water source when rainfalls. Also, Factor 2 is fluctuation of water quality by the geographical characteristics. Therefore, the counterplans against non-point pollution source must be taken. Filtration and disinfection facilities are needed in the areas which have surface water as water source.

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방사선 그라프트 중합법을 이용한 이온교환 필터의 살균효과 (Sterilization Effect of the Ion-exchanger Filter Using by Radiation Graft Polymerization)

  • 김예진;홍용철;김민
    • Korean Chemical Engineering Research
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    • 제52권3호
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    • pp.378-381
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    • 2014
  • 방사선 그라프트 중합에 의한 이온교환필터를 이용하여 물에 포함된 미생물의 살균 효과를 연구하였다. 이온교환필터는 부직포 필터에 전자선을 조사한 후, GMA를 그라프트 중합시켰으며, 이 GMA 필터에 이온교환기를 도입시켜 이온교환필터(EtA, DEA, SS)를 도입하였다. 그 결과 이온교환기 밀도는 EtA의 경우 2.38 mol/kg, DEA는 1.79 mol/kg, SS는 0.75 mol/kg으로 나타났다. 이렇게 제작된 필터를 통해 E. coli의 살균력을 측정하였다. SS-diol 필터의 경우 log 4.65로 EtA, DEA에 비해 각각 약 3.00배, 1.10배 높은 제거율이 나타났다. 이는 상수처리 기준 3000 CFU/ml와 비교하여 우수한 결과를 나타냈음을 알 수 있었다.

다환방향족탄화수소에 대한 오존처리의 방법에 관한 연구 (A Study on the Degradation of PAH in Organic and Aqueous Phases by Ozone)

  • 최영익;손희종;정철우
    • 한국물환경학회지
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    • 제22권6호
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    • pp.1123-1129
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    • 2006
  • 대부분의 polycyclic aromatic hydrocarbons (PAHs) 은 최우선 오염물질로 간주되어지고 있는 매우 유독한 물질이다. Pyrene은 PAHs들 중에서도 그 유독성은 가히 심각하다. 그리고 Pyrene과 다른 PAHs화합물들은 물에 잘 용해가 되지 않는 소수성 성질을 가지고 있어 화학적 또는 생물학적 분해가 용이하지 않다. 이러한 성질을 극복하기 위하여 본 연구에서는 Pyrene을 대표 물질로 하여 2 단계 오존처리를 하였다. 첫 단계에서 Pyrene을 무극성인 핵산 용매에 대량 (2000 mg/L) 으로 녹여 오존처리를 하였다. 이때 Pyrene은 극성 분자들, 즉 알콜과 알데하이드 그리고 에시드 기능기를 가지는 물질들로 변화되었으며 이 변화된 물질들을 다시 극성 용매, 물에 녹여 두 번째 오존처리를 하였다. 두 번째 오존 처리된 Pyrene의 부산물들과 중간생성물들은 생물학적 처리로 가능한지 연구되어지기 위해 $BOD_5$와 COD 그리고 E-coli toxicity tests가 이루어졌다. 그 결과 오존 처리된 Pyrene은 유독하지도 않았고 Pyrene의 부산물들과 중간생성물들은 생물학적 처리가 용이하여졌다. Gas chromatograph (GC) 분석을 통해 Pyrene의 부산물들과 중간생성물들을 밝혀내었다. 이 연구를 토대로 소수성 성질을 가지는 많은 방향족 물질들을 처리하기가 매우 용이해졌다.

Incidence and Management of Toxicity Associated with L-Asparaginase in the Treatment of ALL and NK/T-Cell Lymphoma: an Observational Study

  • Yeang, Shu Hui;Chan, Alexandre;Tan, Chuen Wen;Lim, Soon Thye;Ng, HengJoo
    • Asian Pacific Journal of Cancer Prevention
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    • 제17권7호
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    • pp.3155-3160
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    • 2016
  • Background: L-asparaginase (ASNase) is commonly used in the treatment of acute lymphoblastic leukemia (ALL) and natural killer (NK)/T-cell lymphoma. This study was designed to describe the incidence of toxicity associated with ASNase in Asian adults. Secondary objectives were to investigate the management and impact of toxicity on subsequent ASNase use, and to compare the actual management against current recommendations. Materials and Methods: In this retrospective, multi-center, observational study, Asian patients ${\geq}18$ years old who received ${\geq}1$ dose of the native E. coli ASNase from 2008 to 2013 were included. Patients were excluded if they did not receive ASNase. Endpoints of this study were development of specific toxicities, whether ASNase was discontinued or re-challenged, and developmentg of recurrent toxicity. All data analyses were performed using SPSS version 20.0. Results: A total of 56 patients were analyzed. Mean (${\pm}SD$) age was 36.2 (${\pm}15.2$) years old, with 62.5% being males, 55.4% with ALL and 28.6% with NK/T-cell lymphoma. Hypersensitivity (12.5%) was associated with the highest incidence of toxicity (6 out of 7 patients had Grade 3 and 4 toxicity), followed by 10.7% for hepatic transaminitis, 3.6% for non-CNS thrombosis and 1.8% each for hyperbilirubinemia and pancreatitis. Hypersensitivity recurred in the 3 patients who were re-challenged with E. coli ASNase. Conclusions: ASNase is associated with a wide range of toxicities, with hypersensitivity being the most commonly observed among Asian adult patients.

Syntheses of Biologically Non-Toxic ZnS:Mn Nanocrystals by Surface Capping with O-(2-aminoethyl)polyethylene Glycol and O-(2-carboxyethyl)polyethylene Glycol Molecules

  • Kong, Hoon-Young;Song, Byung-Kwan;Byun, Jonghoe;Hwang, Cheong-Soo
    • Bulletin of the Korean Chemical Society
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    • 제34권4호
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    • pp.1181-1187
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    • 2013
  • Water-dispersible ZnS:Mn nanocrystals were synthesized by capping the surface of the nanocrystal with O-(2-Aminoethyl)polyethylene glycol (PEG-$NH_2$, Mw = 10,000 g/mol) and O-(2-Carboxyethyl)polyethylene glycol (PEG-COOH, Mw = 10,000 g/mol) molecules. The modified PEG capped ZnS:Mn nanocrystal powders were thoroughly characterized by XRD, HR-TEM, EDXS, ICP-AES and FT-IR spectroscopy. The optical properties were also measured by UV/Vis and photoluminescence (PL) spectroscopies. The PL spectra showed broad emission peaks at 600 nm with similar PL efficiencies of 7.68% (ZnS:Mn-PEG-NH2) and 9.18% (ZnS:Mn-PEG-COOH) respectively. The measured average particle sizes for the modified PEG capped ZnS:Mn nanocrystals by HR-TEM images were 5.6 nm (ZnS:Mn-PEG-NH2) and 6.4 nm (ZnS:Mn-PEG-COOH), which were also supported by Debye-Scherrer calculations. In addition, biological toxicity effects of the nanocrystals over the growth of wild type E. coli were investigated. They showed no biological toxicity to E. coli until very high concentration dosage of 1 mg/mL of the both nanocrystal samples.