• 제목/요약/키워드: yeast-based assay

검색결과 56건 처리시간 0.02초

Amylocolatosis sp.가 생산하는 항암물질의 정제 및 구강암 모델에 미치는 항암 효과 (Purification of Materials Produced by Amylocolatosis sp. and Anticancer Effect in Oral Cancer Model)

  • 김정;박영민;임도선
    • 치위생과학회지
    • /
    • 제3권1호
    • /
    • pp.11-14
    • /
    • 2003
  • 각종 토양시료로부터 M배지와 AMP배지에서 생육하는 500여종의 방선균을 분리하였으며, 그 중 전배양배지(GMY 배지)에서 가장 뛰어난 항암활성능을 소유한 균주를 불리하고 배양하여 세포외 항암성물질을 분리하여 이 물질을 완전정제하고 MTT 정량분석을 실시하여 암세포에 대한 세포독성검사를 실시하였다. 1. 정제는 배양한 균체를 완전히 제거하고 동량의 ethylacetate를 처리하여 배양액중의 항암성분을 추출하고 무수 magnesium sulfate로 건조 후 농축, ethanol로 용해, 10배량의 water을 첨가하여 $4^{\circ}C$에서 overnight 시킨 후 추출액을 원심분리(12,000 rpm, 30분)하여 methylene chloride로 용해시켜 silica gel 60 column(${\phi}35{\times}600mm$, Merck Co.), methylene chloride-ethanol(96:4) 용매로 용출하고 sephadex LH-20 column(${\phi}15{\times}300mm$, Pharmacia LKB)에서 100% methanol로 용출하여 HPLC(Waters, ODS)를 이용하여 최종적으로 완전 정제하였다. 2. Compound는 Gram(+) 세균(6균주), Gram(-) 세균(11균주), 효모(2균주), 곰팡이(1균주)에 대하여 항균 효과를 나타내었다. 3. 완전 정제된 물질에 대한 항암효과를 측정한 결과, 모든 실험 암세포에 대해서 뛰어난 항암효과를 나타내었으며 protein성 물질은 아닌 것으로 추정되었다.

  • PDF

Discovering Novel Genes of poultry in Genomic Era

  • S.K. Kang;Lee, B.C.;J.M. Lim;J.Y. Han;W.S. Hwang
    • 한국가금학회지
    • /
    • 제28권2호
    • /
    • pp.143-153
    • /
    • 2001
  • Using bioinformatic tools for searching the massive genome databases, it is possible to Identify new genes in few minutes for initial discoveries based on evolutionary conservation, domain homology, and tissue expression patterns, followed by further verification and characterization using the bench-top works. The development of high-density two-dimensional arrays has allowed the analysis of the expression of thousands of genes simultaneously in the humans, mice, rats, yeast, and bacteria to elucidate the genes and pathways involved in physiological processes. In addition, rapid and automated protein identification is being achieved by searching protein and nucleotide sequence databases directly with data generated from mass spectrometry. Recently, analysis at the bio-chemical level such as biochemical screening and metabolic profiling (Biochemical genomics) has been introduced as an additional approach for categorical assignment of gene function. To make advantage of recent achievements in computational approaches for facilitated gene discoveries in the avian model, chicken expression sequence tags (ESTs) have been reported and deposited in the international databases. By searching EST databases, a chicken heparanase gene was identified and functionally confirmed by subsequent experiments. Using combination of sub-tractive hybridization assay and Genbank database searches, a chicken heme -binding protein family (cSOUL/HBP) was isolated in the retina and pineal gland of domestic chicken and verified by Northern blot analysis. Microarrays have identified several host genes whose expression levels are elevated following infection of chicken embryo fibroblasts (CEF) with Marek's disease virus (MDV). The ongoing process of chicken genome projects and new discoveries and breakthroughs in genomics and proteomics will no doubt reveal new and exciting information and advances in the avian research.

  • PDF

The effect of nitrogen-fixing microorganisms on plant promotion in cabbage

  • Moon, Je-Hun;Jadamba, Chuluuntsetseg;Yoo, Soo-Cheul
    • 한국작물학회:학술대회논문집
    • /
    • 한국작물학회 2017년도 9th Asian Crop Science Association conference
    • /
    • pp.190-190
    • /
    • 2017
  • Chemical fertilizers have been used to increase crop production and contributed to escaping food shortages. However, excessive use of chemical fertilizers over a long period caused many problems such as environmental pollution and the hampered production potential of the land. Thus, it is necessary to develop eco-friendly bio-fertilizers that can replace the use of chemical fertilizers. Here, we tested the effect of some nitrogen-fixing microorganims on the plant growth promotion. Seventy free-living nitrogen fixing microorganisms were isolated from rhizosphere of crop cultivation fields, streamside soils and sludge in Ansung, Korea. Of them, three strains (NF2-4-1, Yeast; EMM409, Mesorhizobium; Gsoil662, Burkholderia) were selected to be most efficient in the capacity of N-fixing nitrogen based on colony forming cell assay in N-free media. To investigate the ability to promote plant growth, these strains were inoculated into the soil and cabbage were grown for 4 weeks in the grown chamber. Fresh weight, dry weight, and leaf area were measured from 4-week-old plants. Phenotypic analysis revealed that the growth of the plants inoculated with NF2-4-1 and EMM409 strains were significantly promoted compared to the mock-treated control plants, while Gsoil662-inoculated plants did not show statically significant promotion. These results indicate that these nitrogen-fixing microorganims can be used to develop plant growth promoting bio-fertilizers. Further analysis on nitrogen fixing level in soil by these strains will be tested.

  • PDF

음식물 쓰레기로부터 Protease를 생산하는 B. amyloliquefaciens JH-35의 분리 및 특성 (Isolation and Characterization of Protease Producing B. amyloliquefaciens JH-35 from Food Waste)

  • 유재홍;주진호;김성국;장인환
    • 한국환경농학회지
    • /
    • 제35권4호
    • /
    • pp.294-301
    • /
    • 2016
  • BACKGROUND: Recent studies have described the importance of microbes and enzymes that can compost food waste. This study was carried out to improve production of protease of isolated microbes from food waste. METHODS AND RESULTS: Seven bacteria isolated from various sources were screened for protease production by adding skim milk into the agar medium. About 7 microbes producing protease were tested, and strain JH-35 showed the highest protease activity among them. The strain was identified as Bacillus amyloliquefaciens JH-35 based on morphological, cultural, physiological characteristics and 16S rRNA. In the fermentation experiment, the assay B. amyloliquefaciens JH-35 showed the highest protease activity in the condition of 1% glucose, 1.5% yeast extract and 0.2%$ K_2HPO_4$. The optimal condition of culture with temperature $35^{\circ}C$, initial pH of 7 and shaking speed of 200 rpm and 24 hr. CONCLUSION: The protease of the B. amyloliquefaciens JH-35 had its activity at pH 7 and the optimal culture time was 24 hr. Also, B. amyloliquefaciens JH-35 was high salt tolerance. Our results suggest that B. amyloliquefaciens JH-35 from food waste may have the potential to degrade protein and carbohydrate in food waste.

Effects of Lactobacillus reuteri-based Direct-fed Microbial Supplementation for Growing-Finishing Pigs

  • Shon, K.S.;Hong, J.W.;Kwon, O.S.;Min, B.J.;Lee, W.B.;Kim, I.H.;Park, Y.H.;Lee, I.S.
    • Asian-Australasian Journal of Animal Sciences
    • /
    • 제18권3호
    • /
    • pp.370-374
    • /
    • 2005
  • Two experiments were conducted to evaluate the effects of direct-fed microbial supplementation on the growth performance and nutrient digestibility in pigs. In experiment 1, forty eight pigs were used in a 42-d growth assay. There were four pigs per pen and three pens per treatment. Dietary treatments included 1) NC (without antibiotic basal diet), 2) PC (NC diet+0.1% antibiotic, 100 g/kg chlortetracycline), 3) DFM-1 (NC diet+0.2% Lactobacillus reuteri and Lactobacillus salivarius complex) and 4) DFM-2 (NC diet+0.2% Lactobacillus reuteri, Lactobacillus salivarius, Lactobacillus plantarum and Yeast complex). For the overall period, no treatment had significant effects in growth performance. In the nutrition digestibility, the pigs fed DFM diets were improved in DM and N digestibility compared with the pigs fed NC and PC diets but it was not significantly different. In experiment 2, sixty four crossbred pigs were used in a 98-d growth assay. There were four pigs per pen and four pens per treatment. Dietary treatments included 1) HND (high nutrient diet), 2) LND (low nutrient diet), 3) HND+DFM (HND diet+0.2% Lactobacillus reuteri and Lactobacillus plantarum complex) and 4) LND+DFM (LND diet+0.2% Lactobacillus reuteri and Lactobacillus plantarum complex). For overall period of growing phase, the pigs fed LND diets had improved gain/feed (p=0.01) and for overall period in the finishing phase, the pigs fed LND with DFM diets had higher ADG, ADFI and gain/feed than the others but there were no significant differences (p>0.13). In total period of growing-finishing phase, the pigs fed LND diet had higher gain/feed than the pigs fed HND diets (p<0.05). In growing phase, there were not significant differences among the treatments means for DM and N digestibility. However, the pigs fed diets with DFM had improved N digestibility (p<0.02) compared to the pigs fed diets without DFM in finishing phase. In conclusion, DFM slightly improved the growth performance in growing-finishing pigs.

다기능 탄산칼슘 형성세균의 시멘트 건축물 적용위한 부식능 평가 및 건축물 정주미생물 중 방제 대상 결정 (Assessment of Bio-corrosive Effect and Determination of Controlling Targets among Microflora for Application of Multi-functional CFB on Cement Structure)

  • 박종명;박성진;김사열
    • 생명과학회지
    • /
    • 제25권2호
    • /
    • pp.237-242
    • /
    • 2015
  • 탄산칼슘 형성세균에 의한 건축물 균열보수 및 그들의 내구성 증진은 기존 화학제에 비해 친환경적이며, 영구적이라는 측면에서 그들의 응용에 대한 연구는 중요한 위치를 차지하고 있다. 그러나 균열복구 기전이 세균의 대사에 의한 calcite crystal 형성과정에서 기인되는 것으로, 대사과정 중 산 형성으로 시멘트 기반 콘크리트 건축물에 대한 위해 가능성 논란이 지속 제기되었다. 따라서 본 연구는 시멘트 모르타르에 대한 생물학적 부식능이 있는 방제대상 미생물을 결정하고 시멘트 콘크리트 건축물에의 안전한 적용을 위한 다기능성 탄산칼슘 형성세균을 평가하기 위해 수행되었다. 이를 위해 첫째, 건축물에서 분리된 균사형 진균, 효모, 세균 균주들을 대상으로 chalky test를 실시하여 $CaCO_3$에 대한 가용화능을 평가하였다. 그 결과 건축물에서 분리된 세균 균주의 경우 가용화 능이 없는 것으로 평가되었으며, C. sphaerospermum KNUC253 및 P. prolifica KNUC263 균주가 $CaCO_3$에 대한 가용화 활성을 보여, 시멘트 기반 건축물 표면에서 방제되어야 균주로 판단된다. 둘째, 독도 토양 및 건축물에서 분리되어 시멘트 표면 진균생장 억제효과, 시멘트 표면균열을 수복 및 수분 침투력 감소효과가 보고된 특허등록 균주 A. nicotianae KNUC2100, B. thuringiensis KNUC2103, S. maltophilia KNUC2100 및 B. aryabhatti KNUC205들이 $CaCO_3$을 가용화하지 않음을 볼 때 산 형성으로 인한 시멘트 모르타르 주성분인 $CaCO_3$에 대한 위해는 없을 것으로 평가된다. 이에 더하여 그들 중 B. aryabhatti KNUC205, A. nicotianae KNUC2100의 경우 건축소재 열화가능성의 판단근거로 널리 활용되고 있는 bio-degradative assay에서 다양한 고분자물질 구성요소에 대해 활성을 나타내지 않아 다양한 건축환경에 대해 활용 가능성을 보여주었다.