• 제목/요약/키워드: Selective isolation

검색결과 171건 처리시간 0.031초

${\beta}-Glucuronidase$ 생산 인체장내 Clostridium sp.의 분리 . 동정 (Isolation and Identification of ${\beta}-Glucuronidase$ producing Clostridium sp. from Fecal Microflora)

  • 박종현;신지영
    • 한국식품과학회지
    • /
    • 제31권5호
    • /
    • pp.1357-1362
    • /
    • 1999
  • 한국인의 장내세균중의 유해효소생성 균주를 분리하여 장내환경 개선연구에 활용하고자 ${\beta}-glucuronidase$$7{\alpha}-dehydroxylase$의 유해효소를 분비하는 장내세균으로 알려진 Clostridium속 균주를 분리하였다. 동일인의 장내세균을 2회에 걸쳐서 검색한 후 최다 우점종이외의 장내세균은 동일인이라도 식이등 외부환경에 의하여 변화될 수 있음을 알았다. 이어서 Neomycin-Nagler 선택배지를 사용하여 Clostridium spp.를 14종 분리하여 이들 균주가 유해효소를 생산하는지를 조사하였다. 일차 선발균주중 Clostridium sp. Isolate-11은 ${\beta}-glucuronidase$를 강력히 생성하여 0.021 unit/mg protein의 활성을 보여주었으나 이들 분리 균주중에서 $7{\alpha}-dehydroxylase$의 역가를 TLC상에서 검출할 수 있는 균주는 발견하지 못했다. Isolate-11를 배양생리학적으로 동정한 결과 이 균은 Clostridium scatologenes인 것으로 잠정동정하였다.

  • PDF

Isolation and Biological Properties of Novel Cell Cycle Inhibitor, HY558, Isolated from Penicillium minioluteum F558

  • Lee, Chul-Hoon;Lim, Hae-Young;Kim, Min-Kyoung;Cho, Youl-Hee;Oh, Deok-Kun;Kim, Chang-Jin;Lim, Yoon-Gho
    • Journal of Microbiology and Biotechnology
    • /
    • 제12권3호
    • /
    • pp.470-475
    • /
    • 2002
  • In the course of screening for a novel cell cycle inhibitor, a potent Cdk 1 inhibitor, HY558, was found from the culture broth of Penicillium minioluteum F558 isolated from a soil sample. The molecular ion of HY558 was identified at m/z 329 (MH+) with a molecular formula of $C_20H_44ON_2$. HY558 exhibited selective antiproliferative effects on various human cancer cell lines. Its $IC_50$ values were estimated to be 0.29 mM on HepG2, 0.30 mM on HeLa, 0.30 mM on HL6O, 0.33 mM on HT-29, and 0.25 mM on AGS cells. Interestingly, Hy558 demonstrated no antiproliferative effect with normal lymphocytes used as the control, and a low level of inhibition on the proliferation of A549 cancer cells. A flow cytometric analysis of HepG2 cells revealed an appreciable arrest of cells at the G1 and G2/M phases of the cell cycle following treatment with Hy558. furthermore, DNA fragmentation due to apoptosis was observed in HeLa cells treated with 0.46 mM of HY558.

Isolation and Characterization of Bifidobacterium longum subsp. longum BCBR-583 for Probiotic Applications in Fermented Foods

  • Yi, Da Hye;Kim, You-Tae;Kim, Chul-Hong;Shin, Young-Sup;Lee, Ju-Hoon
    • Journal of Microbiology and Biotechnology
    • /
    • 제28권11호
    • /
    • pp.1846-1849
    • /
    • 2018
  • Recent human gut microbiome studies have supported that the genus Bifidobacterium is one of the most beneficial bacteria for human intestinal health. To develop a new probiotic strain for functional food applications, fourteen fecal samples were collected from healthy Koreans and the strain BCBR-583 was newly selected and isolated from a 25-year-old Korean woman's fecal sample using the selective medium for Bifidobacterium. Subsequent fructose-6-phosphate phosphoketolase (F6PPK) test and 16S rRNA gene sequencing analysis of the strain BCBR-583 confirmed that it belongs to B. longum subsp. longum. The stress resistance tests showed that it has oxygen and heat tolerance activities (5- and 3.9-fold increase for 24 h at 60 and 120 rpm, respectively; $78.61{\pm}6.67%$ survival rate at $45^{\circ}C$ for 24 h). In addition, gut environment adaptation tests revealed that this strain may be well-adapted in the gut habitat, with gastric acid/bile salt resistance ($85.79{\pm}1.53%$, survival rate under 6 h treatments of gastric acid and bile salt) and mucin adhesion ($73.72{\pm}7.36%$). Furthermore, additional tests including cholesterol lowering assay showed that it can reduce $86.31{\pm}1.85%$ of cholesterol. Based on these results, B. longum BCBR-583 has various stress resistance for survival during food processing and environmental adaptation activities for dominant survival in the gut, suggesting that it could be a good candidate for fermented food applications as a new probiotic strain.

Isolation, Identification and Characterization of Phytophthora katsurae, Causing Chestnut Ink Disease in Korea

  • Lee, Jong-Kyu;Jo, Jong-Won;Shin, Keum-Chul;Lee, Sang-Hyun;Lee, Sang-Yong
    • The Plant Pathology Journal
    • /
    • 제25권2호
    • /
    • pp.121-127
    • /
    • 2009
  • Since July 2005, survey of chestnut ink disease was carried out in chestnut stands located at southern parts of Korea. Dead chestnut trees showing inky ooze on necrotic trunks were found in two different locations. In order to isolate and identify the causal fungus, infected tissues and soil samples around dead or dying trees were collected and placed on Phytophthora-selective medium. Rhododendron and chestnut tree leaves were used as a bait to isolate the fungus from soil samples by attracting zoospores in soil suspensions. On V-8 culture medium, the isolates produced homothallic oogonia with protuberances ($34.0-46.2{\times}21.9-26.7{\mu}m$) abundantly, but did not produced sporangia. Mass production of sporangia was possible by immersing agar plugs with actively growing mycelium in the creek water at $18^{\circ}C$ for 3 days. Sporangia were papillate, and ovoid to obpyriform ($17.0-38.9{\times}14.6-29.2{\mu}m$) in shape. Comparison of the ITS sequences revealed that the isolates had 100% identity to the P. katsurae isolates from Japan and New Zealand and 99.6% identity to other P. katsurae isolates. All of the examined isolates from Korea were completely identical to each other in ITS sequence. Numerous sporangia were formed in filtered as well as unfiltered creek water, but no sporangia formed in sterilized distilled water. Light induced sporangia formation, but has no influence on oospore formation. Amendments of ${\beta}$-sitosterol in culture media have no significant effect on mycelial growth but significantly stimulate oospore and sporangia formation.

Chitinase를 생성하는 Serratia sp. JM의 분리 및 특성 (Isolation and Characterization of Serratia sp. JM Producing Chitinase)

  • 차진명;진상기고한철이인화
    • KSBB Journal
    • /
    • 제11권1호
    • /
    • pp.92-98
    • /
    • 1996
  • 전남 법성포 해안의 갯벌 시료로부터 chitinase 생 생 균주를 분리하였으며, 분리된 균주 중에서 chiti­n nase 생성능이 우수한 JM을 선발 동정하여 Serranasetia sp. JM으로 명명하였다. Serratia sp. JM은 nu­trient 배지냐 MacConkey 배지에서 prodigiosin 색소를 생성하며, 정제 chitin이 포함된 한천 배지에서 는 chitinase 생성에 따른 clear zone 형성이 확인되 었다 Serratia sp. JM은 형태적, 생리.생화학적 특 성과 유기물 동화는 SUCCIniC, urea 및 pyruvic 산 을 제외하고는 공시 균주인 Serratia marcescens ATCC 27117과 유사하였으며, tetracyclin에 대해 서는 항생제에 대한 내성을 가지고 있었으나, kanamycin과 chloramphenicol에 대해서는 내성을 가지지 않았다. Serratia sp. JM의 chitinase 생성에 따른 최적온도와 pH는 $30^{\circ}C$ 와 7.5로 냐타났다. Serratia sp. JM은 120시간까지는 배양 시간이 증가 함에 따라 chiti-nase 생성과 pH는 점차 증가하였으 나, 배양 120시간 이후에는 chitin 분해에 따른 acetic acid의 축적에 따라 chitinase 생성과 pH는 감소 하였다.

  • PDF

Generation of Anti-HLA-DR4 Specific Antibodies by Immunization of the Recombinantly Expressed Allelic Subtype-Specific Region of the $HLA-DRB1^*0405$ Molecules

  • Park, Jung-Hyun;Cho, Eun-Wie;Lee, Yun-Jung;Chung, Jin;Hahm, Kyung-Soo;Kim, Kil-Lyong
    • BMB Reports
    • /
    • 제31권2호
    • /
    • pp.111-116
    • /
    • 1998
  • HLA-DR4 is the dominant allele of MHC class II genes in Koreans. In particular, the $DRB1^*0405$ subtype has been reported to be almost exclusively expressed in Far East Asians, and has also been observed to be strongly associated with rheumatoid arthritis in Koreans and the Japanese. Identification of this specific allele has been mainly performed by PCR-based methods, which is often time consuming, costly, and involves tedious procedures such as the isolation of genomic DNA, PCR, and gel electrophoresis. To develop a more convenient tool for screening vast amounts of samples as well as to generate reagents which might also be used in other applications, in this study, antibodies were produced against this specific HLA subtype. By PCR, an allelespecific region covering the ${\beta}1$ domain of $DRB1^*0405$ was amplified and recombinantly expressed in E.coli. Immunization of Lewis rats with the purified protein yielded an allele specific antiserum. Western blot analysis showed the selective detection of the HLA-DR ${\beta}-chain$. Using this antiserum, established cell lines and peripheral blood lymphocytes were analyzed on their HLA haplotype by fluorescence activated flow cytometry. These novel antibodies will provide a powerful tool in the detection and investigation of DR4 alleles.

  • PDF

Shigellosis

  • Niyogi Swapan Kumar
    • Journal of Microbiology
    • /
    • 제43권2호
    • /
    • pp.133-143
    • /
    • 2005
  • Shigellosis is a global human health problem. Four species of Shigella i.e. S. dysenteriae, S. flexneri, S. boydii and S. sonnei are able to cause the disease. These species are subdivided into serotypes on the basis of O-specific polysaccharide of the LPS. Shigella dysenteriae type 1 produces severe disease and may be associated with life-threatening complications. The symptoms of shigellosis include diarrhoea and/or dysentery with frequent mucoid bloody stools, abdominal cramps and tenesmus. Shigella spp. cause dysentery by invading the colonic mucosa. Shigella bacteria multiply within colonic epithelial cells, cause cell death and spread laterally to infect and kill adjacent epithelial cells, causing mucosal ulceration, inflammation and bleeding. Transmission usually occurs via contaminated food and water or through person-to-person contact. Laboratory diagnosis is made by culturing the stool samples using selective/differential agar media. Shigella spp. are highly fragile organism and considerable care must be exercised in collecting faecal specimens, transporting them to the laboratories and in using appropriate media for isolation. Antimicrobial agents are the mainstay of therapy of all cases of shigellosis. Due to the global emergence of drug resistance, the choice of antimicrobial agents for treating shigellosis is limited. Although single dose of norfloxacin and ciprofloxacin has been shown to be effective, they are currently less effective against S. dysenteriae type 1 infection. Newer quinolones, cephalosporin derivatives, and azithromycin are the drug of choice. However, fluoroquinolone-resistant S. dysenteriae type 1 infection have been reported. Currently, no vaccines against Shigella infection exist. Both live and subunit parenteral vaccine candidates are under development. Because immunity to Shigella is serotype-specific, the priority is to develop vaccine against S. dysenteriae type 1 and S. flexneri type 2a. Shigella species are important pathogens responsible for diarrhoeal diseases and dysentery occurring all over the world. The morbidity and mortality due to shigellosis are especially high among children in developing countries. A recent review of literature (KotIoff et al.,1999) concluded that, of the estimated 165 million cases of Shigella diarrhoea that occur annually, $99\%$ occur in developing countries, and in developing countries $69\%$ of episodes occur in children under five years of age. Moreover, of the ca.1.1 million deaths attributed to Shigella infections in developing countries, $60\%$ of deaths occur in the under-five age group. Travellers from developed to developing regions and soldiers serving under field conditions are also at an increased risk to develop shigellosis.

가두리 양식장의 Vibrio vulnificus 검출 및 제어 방법 (Detection of Vibrio vulnificus in Fish Farm and Bactericidal Methods on this Bacteria)

  • 성치남;송계민;이규호;양성렬
    • 미생물학회지
    • /
    • 제38권4호
    • /
    • pp.281-286
    • /
    • 2002
  • 2000년 1 월부터 2000년 10월까지 가두리 양식장에서 Vibrio vulnificus를 검출하였고 이들의 억제 방법을 연구하였다. 이 세균의 검출은 선택적 분리법과 vvhA 유전자를 확인하는 방법을 이용하였다. V. vulnificus는 수온이 $17^{\circ}C$이상인 5월부터 검출되었고 $19^{\circ}C$ 이상인 6월부터 9월까지는 대부분의 시료에서 검출되었다. V. vulnificus를 제어하기 위한 방법 중 냉동 및 냉장 처리는 살균효과를 나타내지 못했다. Citric acid도 균의 생장을 억제하지 못했으나, 500 mg/1이상의 EDTA가 첨가될 경우 균이 완전히 사멸되었다. 분말 광촉매인 산화티타니움은 자외선을 조사할 경우 15분~1 시간 이내에 이 세균을 완전히 사멸시키는 효과를 나타내었다. 산화티타니움을 유리 구슬에 코팅한 광촉매 장치를 이용하여 0.2/min의 turnover rate로 사멸효과를 얻었다.

Analysis of cellular fatty acid methyl esters (FAMEs) for the identification of leuconostoc strains isolated from kimchi

  • Lee, Jung-Sook;Chun, Chang-Ouk;Kim, Hong-Joong;Joo, Yun-Jung;Lee, Hun-Joo;Park, Chan-Sum;Park, Yong-Ha;Mheen, Tae-Ick
    • Journal of Microbiology
    • /
    • 제34권3호
    • /
    • pp.225-228
    • /
    • 1996
  • The cellular fatty acid methyl esters (FAMEs) analysis data obtained for clusters defined at a Euclidian distance of 17.5, in the classification of lactic acid bacteria isolated from kimchi, described by Lee et al. (4), was used for the identification of 79 Leuconostoc isolates. The test strains were isolated using a selective isolation medium specific for the genus Leuconostoc. These strains were then characterized according to their fatty acid profiles. The results show that all seventy nine test strains were identified to the known Leuconostoc clusters B, C, and D. Cluster B had the highest relative amount of the saturated fatty acid 16 : 0. The saturated fatty acid 16 : 0 and summed feature 9 were found as a major components in cluster C, which had a higher level of summed feature 9 than cluster B. Cluster D is characterized by the highest relative amount of the unsaturated fatty acid 18 : 1 w9c. It is suggested that FAMEs analysis can be successfully applied in the identification of lactic acid bacteria isolated from kimchi.

  • PDF

An Aptamer-Based Electrochemical Sensor That Can Distinguish Influenza Virus Subtype H1 from H5

  • Lee, Jin-Moo;Kim, JunWon;Ryu, Ilhwan;Woo, Hye-Min;Lee, Tae Gyun;Jung, Woong;Yim, Sanggyu;Jeong, Yong-Joo
    • Journal of Microbiology and Biotechnology
    • /
    • 제27권11호
    • /
    • pp.2037-2043
    • /
    • 2017
  • The surface protein hemagglutinin (HA) mediates the attachment of influenza virus to host cells containing sialic acid and thus facilitates viral infection. Therefore, HA is considered as a good target for the development of diagnostic tools for influenza virus. Previously, we reported the isolation of single-stranded aptamers that can distinguish influenza subtype H1 from H5. In this study, we describe a method for the selective electrical detection of H1 using the isolated aptamer as a molecular probe. After immobilization of the aptamer on Si wafer, enzyme-linked immunosorbent assay (ELISA) and field emission scanning electron microscopy (FE-SEM) showed that the immobilized aptamer bound specifically to the H1 subtype but not to the H5 subtype. Assessment by cyclic voltammetry (CV) also demonstrated that the immobilized aptamer on the indium thin oxide-coated surface was specifically bound to the H1 subtype only, which was consistent with the ELISA and FE-SEM results. Further measurement of CV using various amounts of H1 subtype provided the detection limit of the immobilized aptamer, which showed that a nanomolar scale of target protein was sufficient to produce the signal. These results indicated that the selected aptamer can be an effective probe for distinguishing the subtypes of influenza viruses by monitoring current changes.