• 제목/요약/키워드: Isolation plate

검색결과 173건 처리시간 0.023초

Aromatic hydrocarbon분해세균의 검출과 그 plasmid유전자의 특성 (Isolation of aromatic hydrocarbon-degrading bacteria and genetic characterization of their plasmid genes)

  • 김치경;김종우;김영창;민태익
    • 미생물학회지
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    • 제24권1호
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    • pp.67-72
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    • 1986
  • Aromatic hydrocarbon degrading bacteria were isolated from industrial waste by using an agar plate method. The isolate DY-1 was identified as Acinetobacter sp. and found to utilize phenanthrene as tis sole carbon source. THe bacteria were proved to produce salicylic acid as an intermediate from phenanthrene through naphthalene pathway, when the products in the culture were wxamined by thin-layer chromatography. THe $Phn^+$ genes were found to be involved in two plasmids of about 4 and 40kb which were lost and not detected in the DNA samples prepared from the mitomycin C-cured cells by a gel electrophoretic analysis.

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Haloarcular sp. EH-1에 의한 bacteriophage의 분리 (Isolation of Bacteriophage from Haloarcular sp, EH-1)

  • 정명주
    • 생명과학회지
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    • 제13권4호
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    • pp.505-510
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    • 2003
  • 멸치 젓갈에서 분리된 파아지의 일반적인 성질과 파아지 DNA 서열을 분석한 결과는 다음과 같다. 멸치 젓갈에서 분리된 파아지의 플라그는 대체로 작고 선명한 핀 형태로 직경 0.5∼l.0 mm이었으며, 파아지의 형태는 지름 68 nm의 대칭형 두부와 길이 100 nm의 contractile 미부를 가지고 있었고 base plate가 관찰되었다. 파아지 DNA의 크기는 약 20 Kbp 정도이었으며, DNA 염기서열은 H. salinarium 파아지 hp 32와 52.87%의 상동성을 나타내었으나 보다 정확한 분류를 위해 파아지 DNA의 전체 염기서열에 대한 분석이 이루어져야 할 것이다.

In vivo Antinociceptive and Anti-inflammatory Effect of the Two Triterpenes. Ursolic Acid and 23- Hydroxyursolic Acid. of Cussonia bancoensis

  • Tapondjou L.A;Choi, Jong-Won;Lee, Kyung-Tae;Jung, Hyun-Ju;Park, Hee-Juhn
    • 대한약학회:학술대회논문집
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    • 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
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    • pp.371.1-371.1
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    • 2002
  • Triterpenoids. Ursolic acid (1). 23-hydroxyursolic acid (2). and tormentic acid (3) were obtained by the hydrolysis of BuOH fraction of Cussonia bancoensis extract and further chromatographic isolation to test antinociceptive and anti-inflammatory effect of C. bancoensis (Aratiaceae). Compound 1 and 2 exhibited anti-nociceptive effects, which were determined by acetic acid-induced writhing test and hot plate test. However. the effect of tormentic acid was not significant (omitted)

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Characterization and Identification of Organic Selenium-enriched Bacteria Isolated from Rumen Fluid and Hot Spring Water

  • Dalia, A.M.;Loh, T.C.;Sazili, A.Q.;Jahromi, M.F.;Samsudin, A.A.
    • 한국미생물·생명공학회지
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    • 제45권4호
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    • pp.343-353
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    • 2017
  • In the present study, the isolation of selenium (Se)-enriched bacteria from rumen fluid and hot spring water was carried out. Rumen fluid samples were taken from cannulated goats fed a basal diet and the water samples were collected from Selayang hot spring, Selangor-Malaysia. A total number of 140 Se-tolerant isolates were obtained aerobically using an Se-enriched medium and spread plate technique. All the isolates were initially screened for the ability to transform the Se-containing medium to a red-orange culture using a spectrophotometer. Twenty isolates of dark red-orange medium were selected for a screening of the highest Se-containing protein accumulating strains using the dialysis technique and icp.ms to measure the Se content. Four isolates, identified as Enterobacter cloacae (ADS1, ADS7, and ADS11), and Klebsiella pneumoniae (ADS2) from rumen fluid origin, as well as, one isolate from hot spring water (Stenotrophomonas maltophilia (ADS18)), were associated with the highest biomass organic Se-containing protein when grown in a medium enriched with $10{\mu}g/ml$ sodium selenite. In addition, around $50{\mu}g/100{\mu}g$ of the absorbed inorganic Se was accumulated as an organic form. Organic Se-containing protein in all the selected strains showed antioxidant properties in the range of 0.306 to 0.353 Trolox equivalent antioxidant capacity (TEAC) mg/ml. Therefore, these strains may offer a potential source of organic Se due to their Se-tolerant nature and higher biomass organic to inorganic Se ratio.

대황으로부터 세균성 질염 치료를 위한 항균성 물질의 분리 및 특성 (Isolation and Characterization of an Antibacterial Substance from Rheum palmatum for Treatment of Bacterial Vaginosis)

  • 장지은;강동희;윤재우;김현수
    • KSBB Journal
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    • 제32권2호
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    • pp.133-139
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    • 2017
  • Rheum palmatum has traditionally been used as a preventive agent and medication against fever and infection. The aim of this study was to isolate and characterize an antibacterial substance from R. palmatum that is effective against bacterial vaginosis. A methanol extract from R. palmatum showed antibacterial activity against Lactobacillus vaginalis KC TC 3515, Chryseobacterium gleum KCTC 2904, and Sphingomonas paucimobilis KCTC 2834, which cause bacterial vaginosis. After extraction and pH control of the methanol extract from R. palmatum, we found that acidic and alkaline extracts did not show antibacterial activity. A neutral extract (50 mg/mL) displayed an inhibitory zone of 18 mm on a nutrient agar plate with C. gleum KCTC 2904. Fractions No. 11 and 12 among 41 fractions obtained by silica gel column chromatography produced inhibitory zones of 10 mm on nutrient agar plates with C. gleum KCTC 2904. $R_f0.15$ and $R_f0.17$ spots produced by TLC of fraction No. 11 showed antibacterial activity against C. gleum KCTC 2904. Isolation and purification of the peak at a retention time (Rt) of 9.427 min was achieved by HPLC of $R_f0.29spots$. The peak at Rt 9.427 min showed antibacterial activity against C. gleum KCTC 2904.

Polymer Film-Based Screening and Isolation of Polylactic Acid (PLA)-Degrading Microorganisms

  • Kim, Mi Yeon;Kim, Changman;Moon, Jungheun;Heo, Jinhee;Jung, Sokhee P.;Kim, Jung Rae
    • Journal of Microbiology and Biotechnology
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    • 제27권2호
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    • pp.342-349
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    • 2017
  • Polylactic acid (PLA) has been highlighted as an alternative renewable polymer for the replacement of petroleum-based plastic materials, and is considered to be biodegradable. On the other hand, the biodegradation of PLA by terminal degraders, such as microorganisms, requires a lengthy period in the natural environment, and its mechanism is not completely understood. PLA biodegradation studies have been conducted using mainly undefined mixed cultures, but only a few bacterial strains have been isolated and examined. For further characterization of PLA biodegradation, in this study, the PLA-degrading bacteria from digester sludge were isolated and identified using a polymer film-based screening method. The enrichment of sludge on PLA granules was conducted with the serial transference of a subculture into fresh media for 40 days, and the attached biofilm was inoculated on a PLA film on an agar plate. 3D optical microscopy showed that the isolates physically degraded the PLA film due to bacterial degradation. 16S rRNA gene sequencing identified the microbial colonies to be Pseudomonas sp. MYK1 and Bacillus sp. MYK2. The two isolates exhibited significantly higher specific gas production rates from PLA biodegradation compared with that of the initial sludge inoculum.

Application of Molecular Methods for the Identification of Acetic Acid Bacteria Isolated from Blueberries and Citrus Fruits

  • Gerard, Liliana Mabel;Davies, Cristina Veronica;Solda, Carina Alejandra;Corrado, Maria Belen;Fernandez, Maria Veronica
    • 한국미생물·생명공학회지
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    • 제48권2호
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    • pp.193-204
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    • 2020
  • Sixteen acetic acid bacteria (AAB) were isolated from blueberries and citric fruits of the Salto Grande region (Concordia, Entre Rios, Argentina) using enrichment techniques and plate isolation. Enrichment broths containing ethanol and acetic acid enabled maximum AAB recovery, since these components promote their growth. Biochemical tests allowed classification of the bacteria at genus level. PCR-RFLP of the 16S rRNA and PCR-RFLP of the 16S-23S rRNA intergenic spacer allowed further classification at the species level; this required treatment of the amplified products of 16S and 16S-23S ITS ribosomal genes with the following restriction enzymes: AluI, RsaI, HaeIII, MspI, TaqI, CfoI, and Tru9I. C7, C8, A80, A160, and A180 isolates were identified as Gluconobacter frateurii; C1, C2, C3, C4, C5, C6, A70, and A210 isolates as Acetobacter pasteurianus; A50 and A140 isolates as Acetobacter tropicalis; and C9 isolate as Acetobacter syzygii. The bacteria identified by 16S rRNA PCR-RFLP were validated by 16S-23S PCR-RFLP; however, the C1 isolate showed different restriction patterns during identification and validation. Partial sequencing of the 16S gene resolved the discrepancy.

Lactobacillus acidophilus가 생산한 항균물질(抗菌物質)에 관한 연구(硏究) (Studies on antimicrobial agent produced by lactobacillus acidophilus)

  • 김동신
    • 대한수의학회지
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    • 제24권2호
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    • pp.149-162
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    • 1984
  • The research was conducted(1) to confirm the agent(s) responsible for the antimicrobial activity contained in the fermented tomato juice with L. acidophilus(2) to extract and purify the antimicrobial agent(s)(3) to find the biological, physical and chemical properties of the agent(s). The following results were obtained and summarized as followings; 1. The agent responsible for the inhibitory activity was confirmed by both well assay method using fermented tomato juice with L. acidophilus and turbidimetric technique using the cell-free filtrate or neutralized filtrate of tomato acidohilus culture and found exerted antimicrobial agent other than lactic acid. 2. The procedures of purification : The isolation and purification of antimicrobial agent from the lyophilized acidophilus tomato culture were carried out by (1) methanol extraction (2) acetone extraction, (3) Sephadex G-50 gel filtration (4) paper chromatography and (5) thin layer chromatography. 3. The biological, physical and chemical properties of antimicrobial agent: The biological, physical, chemical properties of the purified antimicrobial agent were: (1) The antimicrobial activity was strong against test organisms; Bacillus subtilis(ATCC 6633), Escheichia coli(ATCC 25922), Staphylococcus aureus(ATCC 167), Pseudomonas fluorescens(KFCC 32394), Proteus vulgaris and Shigella dysenteriae. (2) The pH value of the agent was 2.0 and the inhibitory activity was lost when it was neutralized at 7.0 of pH and the agent was heat stable at $121^{\circ}C$ for 60 minutes. (3) The ultraviolet light absorption spectra of methanol-acetone extract and TLC fraction exhibited a maximum absorption at 260nm and 224nm respectively. (4) The most purified agent from TLC plate increased about 130-fold in activity. (5) The agent isolated from TLC plate was free from $H_2O_2$ or lactic acid. 4. Bioautographic assy: By means of bioautography of the agent on silica gel of TLC plate a strong inhibitory activity against B. subtilis was demonstrated. 5. Mass spectrometry: The agent obtained from TLC plate was analyzed by mass spectrometry which show the parent peak at m/e 264 suggesting the molecular weight of the compound and molecular group such as [$C_2H{^+}_4$], [CO], [CH=NH], [$C_3{H^}4_7$], [$\begin{array}{rcl}O\\{\parallel}\\CH_3-C\\\end{array}$], [$C_6-H{^+}_{11}$], [$C_5H{^+}_{11}$], [$\begin{array}{rcl}O\\{\parallel}\\C_5H_7-C^+\\\end{array}$] were suggested.

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Sodium dodecylbenzenesulfonate(soft type)를 분해하는 미생물의 분리 및 특성 (Isolation and characterization of sodium dodecylbenzenesulfonate(soft type)-degrading bacteria)

  • 전홍기;안영희;백형석
    • 미생물학회지
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    • 제27권3호
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    • pp.279-284
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    • 1989
  • 부산시내 주택단지의 하수나 토양에서 LAS 분해능이 우수한 균주를 유일한 탄소원으로서 sodium dodecylbenzenesulfonate(SOBS, soft type)를 포함하는 agar plate를 사용하여 분리히였다 이들 분리균중 2균주가. SOBS를 0.1% 함유한 완전배지에서 30$^{\circ}$C 24시간 배양했을 때' SOBS분해율이 최고 59% 정도까지 냐타내였다 긍속이온에 대한 이들 분리 균의 내성조사결과 대부분 $CdCl^{2}$,와 $ZnCl^{2}$에 비교적 내성이 큰 것으로 나타났으며 분리균의 항생제 내성시험에서는 ampicillin 에 대하여 내성을 나타내었다. 특히 분리균중 1개는 ampicillin에 대해 1,550mg/ml 이상에서도 내성을 나타내었으며 이 분리균주는 Klebsiella 속으로 동정되었다. 이 균주는 5kb와 4kb의 2개의 plasmid를 가지고 있었으며 이들은 E. coli C600 plasmic 균주에 형질전환시켰을 때 SDBS분해능을 나타내었으므로 SDBS분해능은 plasmid에 존재하는 유전인자에 의한 것임을 알 수 있었다.

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청국장에서 분리한 혈전용해효소 생산세균의 분리 및 동정 (Isolation and Charaterization of the Fibrinolytic Enzyme Producing Bacterium isolated from Naturally Fermented Chungkookjang)

  • 손병희;오계헌
    • 한국산학기술학회논문지
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    • 제7권3호
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    • pp.476-482
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    • 2006
  • 본 연구의 목적은 상업적으로 사용하기 위하여 혈전용해효소를 생성하는 세균인 MK-15를 분리하고 동정하는 것이다. 먼저 세균 MK-15는 자연 발효된 콩으로부터 농화분리하였으며, 이 세균에 대한 형태학적 및 다양한 생리학적 특성 조사를 실시하였다. MK-15의 배양 상등액으로부터 혈전용해효소 활성을 fibrin plate 방법으로 solid fibrinolytic activity를 측정하였다. 그 결과, 콩배지에서 자란 MK-15의 혈전용해활성은 대조군으로 사용된 plasmin의 혈전용해활성에 비하여 약 2.5배 높았다. 16S rRNA를 분석한 결과 균주 MK-15는 Bacillus subtilis 군과 99.9% 유사성을 나타내었다. 따라서, 이 세균은 Bacillus sp. MK-15로 명명하였으며. 균주 MK-15는 GenBank에 [DQ163021]로 등록하였다.

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