• 제목/요약/키워드: Liquid cultures

검색결과 237건 처리시간 0.025초

Identification of Stenotrophomonas maltophilia LK-24 and its Degradability of Crystal Violet

  • Kim, Jeong-Dong;Yoon, Jung-Hoon;Park, Yong-Ha;Fusako Kawai;Kim, Hyun-Tae;Lee, Dae-Weon;Kang, Kook-Hee
    • Journal of Microbiology and Biotechnology
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    • 제12권3호
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    • pp.437-443
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    • 2002
  • A number of soil and wastewater samples were collected from the vicinity of an effluent treatment plant for the chemical industry. Several microorganisms were screened fur their ability to decolorize the triphenylmethane group of dyes. As a result, a novel crystal violet dye-degrading strain LK-24 was isolated. Taxonomic identification including 16S rDNA sequencing and phylogenetic analysis indicated that the isolate had a $99.5\%$ homology in its 16S rDNA base sequence with Stenotrophomonas maltophilia. The triphenylmethane dye, crystal violet, was degraded extensively by growing cells of Stenotrophomonas maltophilia LK-24 in agitated liquid cultures, although their growth was strongly inhibited in the initial stage of incubation. This group of dyes is toxic, depending on the concentration used. The dye was significantly degraded at a relatively lower concentration, below $100{\mu}g\;ml^-1$, yet the growth of the cells was totally suppressed at a dye concentration of $250{\mu}g\;ml^-1$. The degradation products of crystal violet were identified as 4,4'-bis(dimethylamino)-benzophenone and ${\rho}$-dimethylaminophenol by Gas chromatography-Mass spectrometry. The 4,4'-bis(dimethylamino)-benzophenone was easily obtained in a reasonable yield, as it was not metabolized further by S. maltophilia LK-24; however, the ${\rho}$-dimethylaminophenol was not easily identifiable, as it was further metabolized.

Bacterial Communities of Biofilms Sampled from Seepage Groundwater Contaminated with Petroleum Oil

  • CHO WONSIL;LEE EUN-HEE;SHIM EUN-HWA;KIM JAISOO;RYU HEE WOOK;CHO KYUNG-SUK
    • Journal of Microbiology and Biotechnology
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    • 제15권5호
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    • pp.952-964
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    • 2005
  • The diesel-degrading activities of biofilms sampled from petroleum-contaminated groundwaters in urban subway drainage systems were examined in liquid cultures, and the microbial populations of the biofilms were characterized by denaturing gel gradient electrophoresis (DGGE) and 16S rDNA sequence analysis. Biofilm samples derived from two sites (19 K and 20 K) at subway Station N and Station I could degrade around $80\%$ of applied diesel within 20 and 40 days, respectively, at $15^{\circ}C$, and these results were strongly correlated with the growth patterns of the biofilms. The closest phylogenetic neighbor of a dominant component in the 19 K biofilm was Thiothrix fructosivorans strain Q ($100\%$ similarity). Four dominant strains in the 20 K biofilm were closely related to Thiothrix fructosivorans strain Q ($100\%$ similarity), Thiothrix sp. CC-5 ($100\%$ similarity), Sphaerotilus sp. IF14 ($99\%$ similarity), and Cytophaga-Flexibacter-Bacterioides (CFB) group bacterium RW262 ($98\%$ similarity). Three dominant members in the Station I biofilms were very similar to uncultured Cytophagales clone CRE-PA82 ($91\%$ similarity), Pseudomonas sp. WDL5 ($97\%$ similarity), and uncultured CFB group bacterium LCK-64 ($94\%$ similarity). The microbial components of the biofilms differed depending on the sampling site. This is the first report on the isolation of clones highly similar to Thiothrix fructosivorans and Thiothrix sp. from biofilms in petroleum-polluted groundwaters, and the first evidence that these organisms may play major roles in petroleum degradation and/or biofilm-development.

Genetic Characterization of Two S-Adenosylmethionine-induced ABC Transporters Reveals Their Roles in Modulations of Secondary Metabolism and Sporulation in Streptomyces coelicolor M145

  • Shin, Su-Kyoung;Park, Hyun-Suh;Kwon, Hyung-Jin;Yoon, Hyun-Jin;Suh, Joo-Won
    • Journal of Microbiology and Biotechnology
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    • 제17권11호
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    • pp.1818-1825
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    • 2007
  • S-Adenosylmethionine (SAM) was previously documented to activate secondary metabolism in a variety of Streptomyces spp. and to promote actinorhodin (ACT) and undecylprodigiosin (RED) in Streptomyces coelicolor. The SAM-induced proteins in S. coelicolor include several ABC transporter components (SCO5260 and SCO5477) including BldKB, the component of a well-known regulatory factor for differentiations. In order to assess the role of these ABC transporter complexes in differentiation of Streptomyces, SCO5260 and SCO5476, the first genes from the cognate complex clusters, were individually inactivated by gene replacement. Inactivation of either SCO5260 or SCO5476 led to impaired sporulation on agar medium, with the more drastic defect in the SCO5260 null mutant (${\Delta}SCO5260$). ${\Delta}SCO5260$ displayed growth retardation and reduced yields of ACT and RED in liquid cultures. In addition, SAM supplementation failed in promoting the production of ACT and RED in ${\Delta}SCO5260$. Inactivation of SCO5476 gave no significant change in growth and production of ACT and RED, but impaired the promoting effect of SAM on ACT production without interfering with the effect on RED production. The present study suggests that SAM induces several ABC transporters to modulate secondary metabolism and morphological development in S. coelicolor.

배양 조건의 변화가 미생물에 의한 석탄의 액화에 미치는 영향 (Effects of Culture Condition on Solubilization of Coal by Microorganisms)

  • 이현호;신현재양지원
    • KSBB Journal
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    • 제11권4호
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    • pp.462-469
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    • 1996
  • 미생물을 이용한 석탄의 액화 연구를 통하여 몇가지 중요한 결론을 얻을 수 있었다. 호주산 갈탄의 효과적인 전처리제로는 HNOJ가 선정되었으며, S. viridosporus와 P. cocos의 최적 초기 pH는 각각 7.5와 4.5로 나타났으며, S. viridosporus가 초기액화 속도 및 전체 액화능에서 P. cocos 보다 우수한 액화능을 보였다. 액화능을 높이기 위한 첨가제로서는 S S. viridosporus는 요소가 효과적 이었고, p. cocos는 랩톤과 트립톤이 효과척으로 나타났다. 최고 액화능은 회분식 교반기 조업에서 S. viridosporus에서 85 % % (w/w)까지 향상시킬 수가 있었다. 액화 기간은 3.5 일까지 단축할 수가 있었다. 분광학적 분석 결과 및 각 단계별로 석탄의 성분을 분석한 결과에서 미생물에 의한 석탄의 액화는 석탄의 계속적 인 산화현 상에 기인함을 밝혔다. 또한, S. viridosporus에 의한 액화의 기작은 미생물의 외분비 물질에 의하여 진행되며, 액화 가작중 80 %는 이차대사생성물에 의한 비효소반응이였으며, 20%는 효소반응으로 나타났다.

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소리쟁이 (Rumex crispus)의 부정근과 모상근 배양에 의한 Anthraquinone 생산성 (Anthraquinone Productivity by the Cultures of Adventitious Roots and Hairy Roots from Curled Dock (Rumex crispus))

  • 장석원;김인현;한태진
    • 식물조직배양학회지
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    • 제26권1호
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    • pp.7-14
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    • 1999
  • 형질전환된 소리쟁이 모상근의 기내 배양시 그 생장 특성과 anthraquinone생산성을 조사하기 위하여 소리쟁이의 모상근을 Agrobacterium rhirogenes strain $A_4$ 접종에 의하여 잎 절편으로부터 유기하여 부정근과 비교하였다. 잎 절편에서의 부정근 형성은 5 $\mu$M NAA를 첨가한 MS배지에서 가장 양호하였다. Paper electrophoresis를 통한 opine분석 결과 모상근에서만 mannopine이 검출되고 부정근에서는 검출되지 않았다. 부정근과 모상근 모두에서 2차근인 측근 근단은 호르몬을 첨가하지 않은 배지에서 신장만 할 뿐 분지하지 않았으나, 1차근인 주근의 근단은 많은 측지를 내며 빠르게 생장하였다. Anthraquinone 함량 증대에 따른 부정근과 모상근의 생장에는 MS기본 배지가 가장 양호하였다. 부정근과 모상근 배양시 생장량과 anthraquinone함량은 5% sucrose를 포함한 0.05 $\mu$M NAA와 0.1 $\mu$M kinetin으로 조성된 MS 액체 배지에서 가장 높은 것으로 나타났으며, 모상근의 anthraquinone함량은 배양 일수에 따라 증가하다가 배양 25일 이후에는 감소하였다.

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황칠나무(Dendropanax morbifera)의 부정근 유도 및 기내증식조건 (Induction and in vitro proliferation of adventitious roots in Dendropanax morbifera)

  • 배기화;김지아;최용의
    • Journal of Plant Biotechnology
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    • 제36권2호
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    • pp.163-169
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    • 2009
  • Dendropanax morbifera (Araliaceae) is an endemic species in Korea and distributed in the southern part of Korea. The roots and stems of this plant have been used for folk medicine for the treatment of migraine headache, dysmenorrheal, and remove wind dampness and for Vanishes production. Production of adventitious roots in D. morbifera by in vitro cultures could be used as alternatives materials. Leaf, stem, and root segments from D. morbifera seedling were cultured on Murashige and Skoog (MS) medium supplemented with 3.0 mg/L IBA and 30 g/L sucrose. After 4 weeks of culture, the highest induction of adventitious roots was obtained from the leaf segment. Frequency of adventitious root formation on medium with various kinds of auxins (IAA, NAA, 2,4-D, and IBA) and various concentrations of IBA (0, 0.1, 0.5, 1.0, 3.0, and 5.0 mg/L) was tested. The maximum induction of adventitious root was obtained on medium with 1.0 mg/L IBA. In liquid culture, growth of root was best 1/2MS medium supplemented with 1.0 mg/L IBA and 30 g/L sucrose. Adventitious roots were cultured in 5 L bioreactor containing 1/2 MS medium supplemented with 1.0 mg/L IBA and 30 g/L sucrose and mass-production of adventitious roots was successfully achieved. This study demonstrated for the first time to produce adventitious roots in D. morbifera.

HPLC에 의한 Phaffia rhodozyma 중의 Astaxanthin의 측정법 (Liquid Chromatographic Determination of Astaxanthin in Phaffia rhodozyma)

  • Xu, Bao-Jun;Fang, Zhe-Ming;Kang, Hyo-Jin;Wang, Qi-Jun;Mo, Eun-Kyoung;Sung, Chang-Keun
    • 한국식품영양학회지
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    • 제16권3호
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    • pp.165-170
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    • 2003
  • Astaxanthin은 수산양식에 필수적인 carotenoid 첨가물일 뿐 아니라 우수한 항암 및 항산화 물질로서 알려져 있다. 현재 astaxanthin은 P. rhodozyma에 의해 생산 할 수 있으나 P. rhodozyma에 함유된 유효성분인 astafanthin의 정량적인 분석 방법이 제시되지 못하였다. 본 연구에서는 astaxanthin에 대한 정확한 정성법과 정량분석법을 제안하였다. 이 방법은HPLC의 역상컬럼을 사용하여 dichlorornethane및 4개의 혼합 용매를 이동상으로 사용하는 것이다. 이 제시된 방법을 활용했을 때 P. rhodozyma 배양액 중의 astaxanthin의 함유량을 정확하게 검출할 수 있다. DMSO 와 acetone의 혼합용매가 최적의 astaxanthin 추출용매이었고 또한 낮은 온도와 어두운 환경 이 최적의 시료처리조건이었다.

Thidiazuron 처리에 의한 히어리나무의 기내번식 (Micorpropagation of Corylopsis coreana by Thidiazuron Treatment)

  • 강효진;문홍규;이재선
    • Journal of Plant Biotechnology
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    • 제30권3호
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    • pp.263-267
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    • 2003
  • This study was conducted to evaluate the effect of thidazuron(TDZ) on shoot proliferation and growth from axillary buds of 20-years-old Corylopsis coreana. Shoots proliferation was effectively achieved on WPM(Woody Plant Medium) supplemented with 0.03∼0.1mg/L TDZ. The highest shoot number(6.5$\pm$0.7) was obtained on 0.1mg/L TDZ treatment. On the TDZ medium shoots formed as clusters less than 1cm in height and therefore needed to subculture on GA$_{3}$ containing medium to induce elongation. In consecutive cultures, phenolic compounds were excreted at the proximal part of the explants and inhibited growth of the explants. Growth inhibition by the compounds was overcome using liquid and paper bridge culture system. About 60% of the elongated shoots rooted on half- strength MS medium containing IBA. Generally, IBA was mire effective on in vitro rooting than NAA with optimal range of 0.5mg/L to 1.0mg/L. Rooted plantlets were transferred in an artificial soil(vermculite) and acclimatized in high humidity greenhouse condition. Survival rate differed greatly depending on rooting types of the explants. Two types of rooting were observed. The first type was direct rooting from the explants. The second type was callus formation followed by rooting from the callus. The explants showing the 1st type rooting survived can be multiplicated in vitro by TDZ treatment followed by elongation with GA$_{3}$ and rooting with IBA.

애기장대(Arabidopsis thaliana) 현탁배양세포의 원형질체로부터 식물체 재분화 (Plant Regeneration from Protoplasts of Suspension Cultured Cells in Arabidopsis thaliana)

  • 김명덕;김준철;진창덕;임창진;한태진
    • 식물조직배양학회지
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    • 제27권2호
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    • pp.125-131
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    • 2000
  • 원형질체는 현탁배양된 세포괴로부터 1% cellulase R-10, 0.25% pectolyase Y-23, 그리고 0.5% driselase가 포함된 CPD 효소용액에서 4시간의 진탕 (40 rpm)조건에서 쉽게 분리되었다. 분리된 원형질체는 1 mg/L 2,4-D, 0.5 mg/L kinetin , 200 rng/L spermidine 그리고 68 g/L glucose가 포함된 KM 액체배지에서 배양되었으며, 또한 현탁배양중인 동종의 feeder cell이 있는 CP배지 위에 액체배양배지를 1 mL균일하게 퍼트린 후,$25^{\circ}C$, 암조건으로 4주간 양육배양하였다. 원형질체 유래 SF 캘러스는 0.05 mg/L IAA, 7 mg/L 2lip와 30mg/L sucrose가 첨가된 MS 재분화 배지에서 광조건으로 4주동안 배양되었을 때, 60% 이상의 shoot를 형성하였다. 이후, 식물의생장조절물질이 첨가되지 않은 MS 배지로 옮겨 뿌리를 유도하였으며, 재분화된 식물체는 토양으로 옮겨 배양 8주후에 종자를 형성하였다.

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Enhanced Lovastatin Production by Solid State Fermentation of Monascus ruber

  • Xu Bao-Jun;Wang Qi-Jun;Jia Xiao-Qin;Sung Chang-Keun
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제10권1호
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    • pp.78-84
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    • 2005
  • The purpose of this study was to optimize the solid state cultivation of Monascus ruber on sterile rice. A single-level-multiple-factor and a single-factor-multiple-level experimental design were employed to determine the optimal medium constituents and to optimize carbon and nitrogen source concentrations for lovastatin production. Simultaneous quantitative analyses of the ${\beta}$-hydroxyacid form and ${\beta}$-hydroxylactone for of lovastatin were performed by the high performance liquid chromatography (HPLC) method with a UV photodiode-array (PDA) detector. The total lovastatin yield ($4{\sim}6\;mg/g$, average of five repeats) was achieved by adding soybean powder, glycerol, sodium nitrate, and acetic acid at optimized levels after 14 days of fermentation. The maximal yield of lovastatin under the optimal composition of the medium increased by almost 2 times the yield observed prior to optimization. The experimental results also indicated that the ${\beta}$-hydroxylactone form of lovastatin (LFL) and the ${\beta}$-hydroxyacid form of lovastatin (AFL) simultaneously existed in solid state cultures of Monascus ruber. while the latter was the dominant form in the middle-late stage of continued fermentation. These results indicate that optimized culture conditions can be used for industrial production of lovastatin to obtain high yields.