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오광근,이철우,전영중,이재홍 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.6
홍색비유황 광합성세균인 Rhodopseudomonas capsulata를 선택하여 packed-bed reactor에서 미생물막을 형성할 때, porous ceramic bead가 다른 담체에 비해 우수하였고, 일정한 유입농도하에서 체류시간(hydraulic retention tiem, HRT)이 짧을수록 미생물막 형성이 양호하였으며, 그 때 반응기내의 세포농도는 11,400mg/l로 현탁처리시의 세포농도에 비하여 3~8배 증가하였다. PBR에서 미생물막의 형성은 cell attachment, microcolony formation, biofilm formation의 단계를 거쳐 형성되는 것으로 관찰되었으며, PBR이 FBR보다 안정적인 미생물 부착을 보였고 특히 PBR에서는 BOD용적 부하가 15gBOD/ℓ·day 이상이 되어도 미생물막의 부착비율은 90% 이상을 유지하였다. 전자현미경으로 담체의 표면 및 내부에 고정화된 광합성세균을 확인할 수 있었다. The formation of microbial films(biofilm) by a non-sulfur phototrophic bacteria, Rhodopseudomonas capsulata, on inorganic media was studied. Porous ceramic beads(PCB) were superior to other immobilizing media for the biofilm formation in a packed-bed reactor. It was found that the formation of microbial films favored a lower hydraulic retention time, showing a higher ratio of cells attatched to the media to those suspended in the solution. The cell concentration in the biofilm reactor was as high as 11,400 mg/ℓ, which is 8-folds of the cell concentration in a ordinary suspended treatment. It was observed that the formation of microbial film by R. capsulata followed a general serial process of cell attachment, microcolony formation, and biofilm formation. The microbial films thus formed was very stable even for an extremely high volumetric BOD loading rate of 15 gBOD/ℓ·day. The scanning electron micrographs of the microbial films showed that the cells were attached to both the surface and pores of the media.
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영지(Ganoderma lucidum) 균사체의 액체배양에 의한 세포외 생물고분자의 생산조건과 특성
이신영,강태수 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.1
새로운 생물산업소재의 탐색 및 개발연구의 일환으로 Ganoderma lucidum(영지1호) 균사체의 액체배양에 의하여 세포외 생물고분자의 생산을 위한 최적 생산조건을 검토하였고, 생성 생물고분자를 분획 정제하여 이의 성분특성을 조사하였다. 세포의 생물고분자 생산을 위한 최적 배양조건은 glucose 5%, yeast extract 0.5%, (NH_4)_2HPO_4 0.1% 및 KH_2PO_4 0.05%(w/v)를 함유한 배지에서 pH 5.0, 온도 30℃ 및 교반속도 100 rpm일 때 얻어졌다. 이들 조건하에서 7일간 플라스크 배양하였을 때, 최대의 균체 및 세포외 생물고분자 농도는 각각 15.2 및 18.8 g/l이었으며, 비생육속도, 비기질소비속도 및 비생산속도는 각각 0.039 hr^-1, 0.043 gg ^-1hr^-1이었다. 세포의 생물고분자는 실온의 물추출에 의하여 수용성 및 물불용성 다당류로 분획되었으며, 두 시료 모두 97% 이상의 당을 함유하였다. 수용성 생물고분자 분획(BWS)과 물불용성 분획(BWI)은 모두 glucose, galactose, mannose, xylose 및 fucose를 함유하였으며, 구성당의 몰비는 각각 3.6 : 1.5 : 2.1 : 0.5: trace 및 2.9 : 3.1 : 2.0 : 1.6 : 0.3이었다. For the screening and the development of the new bio-material, cultural conditions for the exo-biopolymer (EBP) production through the submerged cultivation of Ganoderma lucidum mycelium were investigated. Also, the fractionations and the purifications of the exo-biopolymer were carried out and the chemical compositions of the exo-biopolymer were examined. The optimal culture conditions for the exo-biopolymer production were pH 5.0, 30℃ and 100 rpm of agitation speed in the medium containing of 5% (w/v) glucose, 0.5% (w/v) yeast extract, 0.1% (w/v) (NH_4_)_2HPO_4, and 0.05% (w/v) KH_2PO_4. In the flask cultivation for 7 days under these conditions, the concentration of the maximum exo-biopolymer and the cell mass were 15.4 g/l and 18.8 g/l, respectively. The specific growth rate was 0.039 hr^-1. In addition, the substrate consumption rate, and the exo-biopolymer production rate were 0.043 gg ^-1hr^-1 and 0.025 gg ^-1hr^-1, respectively. The exo-biopolymer was fractionated into BWS (water soluble exo-biopolymer) and BWI (war insoluble exo-biopolymer) by the water insoluble exo-biopolymer) by the water extraction, and the sugar contents fo two fractions were higher than 97% (based on dry basis). The components sugar of BWS and BWI fractions were glucose, galactose, mannose, xylose, and fucose. Their molar ratios were 3.6:1.5:2.1:0.5: trace and 2.9:3.1:2.0:1.6:0.3, respectively.
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( Dhayaalini Bala Gopal ),( Chua Ang Lim ),( Tzar Mohd Nizam Khaithir ),( Jacinta Santhanam ) 한국미생물생명공학회 2017 한국미생물·생명공학회지 Vol.45 No.4
Asymmetric PCR preferentially amplifies one DNA strand for use in DNA hybridization studies. Linear- After-The-Exponential-PCR (LATE-PCR) is an advanced asymmetric PCR method which uses innovatively designed primers at different concentrations. This study aimed to optimise LATE-PCR parameters to pro-duce single-stranded DNA of Candida spp. and Aspergillus spp. for detection via probe hybridisation. The internal transcribed spacer (ITS) region was used to design limiting primer and excess primer for LATE-PCR. Primer annealing and melting temperature, difference of melting temperature between limiting and excess primer and concentration of primers were optimized. In order to confirm the presence of single-stranded DNA, the LATE-PCR product was hybridised with digoxigenin labeled complementary oligonu-cleotide probe specific for each fungal genus and detected using anti-digoxigenin antibody by dot blotting. Important parameters that determine the production of single-stranded DNA in a LATE-PCR reaction are difference of melting temperature between the limiting and excess primer of at least 5℃ and primer con-centration ratio of excess primer to limiting primer at 20:1. LATE-PCR products of Candida albicans, Can-dida parapsilosis, Candida tropicalis and Aspergillus terreus at up to 1:100 dilution and after 1 h hybridization time, successfully hybridised to respective oligonucleotide probes with no cross reactivity observed between each fungal genus probe and non-target products. For Aspergillus fumigatus, LATE-PCR products were detected at 1:10 dilution and after overnight hybridisation. These results indicate high detection sensitivity for single-stranded DNA produced by LATE-PCR. In conclusion, this advancement of PCR may be utilised to detect fungal pathogens which can aid the diagnosis of invasive fungal disease.
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