• Title/Summary/Keyword: Yeast bioassay

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Modification of Estrogenic Effect of Nonylphenol Combined with DEHP in Yeast-based Bioassay (형질전환효모를 이용한 내분비계장애물질검색과 Nonylphenol의 Estrogen 유사작용에 대한 DEHP의 상협작용)

  • 박미선;정해관;박현신;한의식;김종원;엄미옥;정상희;오혜영
    • Toxicological Research
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    • v.17 no.1
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    • pp.65-71
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    • 2001
  • The key targets of endocrine disruptors are nuclear hormone receptors, which bind to steroid hormones and regulate their gene transcription. A yeast-based steroid hormone receptor gene trascription assay was previously developed for the evaluation of chemicals with endocrine modulating activity. The yeast transformants used in this assay contain the human estrogen receptor along with the appropriate steroid response elements upstream of the $\beta$-galactosidase reporter gene. We tried to evaluate several natural and synthetic steroids of their potential to interact directly with the steroid receptor. Some putative endocrine disruptors, including nonylphenol, are weakly estrogenic. But the combined treatment oj these chemicals with di-(2-ethylhexyl)phthalate (DEHP) significantly increased the $\beta$-galactosidase activity in the yeast transformant. These results suggest that we also have to consider the synergistic effects of endocrine disruptors. In this study, we showed that yeast-based bioassay is a valuable tool for screening potential endocrine disruptors and quantitative determination of estrogenicity. And the possibility that the estrogen receptor binds multiple environmental chemicals adds another level of complexity to the interaction between the endocrine disruptors and the human hormone system.

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Multiple-end-point Bioassays Using Microorganisms

  • Iwahashi, Hitoshi
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.6
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    • pp.400-406
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    • 2000
  • Since the 1950s, the numbers of species and chemicals produced have significantly increased. Despite the fact that industrial chemicals have given us numerous benefits, there is no doubt that they have damaged the environment. The chemicals being dispersed on the earth should be carefully controlled to prevent adverse effects. Bioassay is one of the methods to assess chemical safety. In bioassay systems, chemical safety is estimated by monitoring biological responses to environmental pollutants and newly synthesized chemicals. This report introduces multiple-point bioassay systems that are based on chemical sensitivities of microorganisms, responses of one kind of organism, and micro-array technology. Multiple-end-point bioassays enable the prediction of chemicals in the environment and the understanding of toxicities of newly synthesized chemicals.

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A Bioassay of Ginseng Extract s Based on Yeast Growth Determination (효모성장 측정을 이용한 인삼추출물의 생물학적 검정)

  • Jung, Noh-Pal
    • Journal of Ginseng Research
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    • v.5 no.1
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    • pp.24-34
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    • 1981
  • For bioassay of the various extracts of ginseng, the growth determination method using Saccharomyces cerevisiae which was cultured with various doses of the extracts, was studied The water extract, Powder. and ethanol extract were more effective (about 45∼ 110% increase) than saponins or its fractions (about 20∼35% increase). The cold methanol residue showed a increase effect but it was not significant. The bioassay curves for the water extract, ethanol extract, the butanol extracted saponins and the cold methanol- residue were made from the experimental data. From these curves it is possible to find the relation between dose and effectiveness and the optimal doses of various ginseng extracts, and the amount of extract in a sample can be estimated The .angers of sample amount were 0.01% (100ppm) ∼0.32% (3200ppm) fo. the water extract, 0.025% (150ppm)∼0.1% (1000ppm) for the ethanol extract, and 0.008% (80ppm)∼0.016% (160ppm) for the saponins. It was impossible to determine the range for the cold methanol- residue, The acceleration effects on the cell proliferation by a only 0.0008% (8ppm) of the diol- and triol-saponin were measurable in earlier Period (24 hour treatment).

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Laboratory Scale Preparation of S-Adenosyl-L-Methionine from Yeast (효모로부터 S-Adenosyl-L-Methionine의 실험실 규모 생산)

  • 이종남;류양욱;최명언
    • Microbiology and Biotechnology Letters
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    • v.19 no.6
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    • pp.588-591
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    • 1991
  • S-adenosyl-L-methionine (SAM) is essential substrate for biological methylation reactions. The present work describes a reoptimized procedure of SAM preparation in laboratory scale by the method of yeast fermentation. The fermentation medium enriched with methionine and the culture conditions were reoptimized. The isolation steps consisted of 5 steps including extractions, precipitations, and chromatography. This improved procedure over original method provides relatively high yield of biologically active product within a 4 day-period.

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Allelopathic Potential and Substances from Cork Tree (Pbellodendron amurense Rupr.)

  • Park, Young-Goo;Choi, Myung-Suk;Yang, Jae-Kyung;Paik, Ki-Hyon
    • Journal of the Korean Wood Science and Technology
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    • v.29 no.3
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    • pp.92-98
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    • 2001
  • Allelopathic effects of the cork tree (Phellodendron amurense Rupr.) on several crops and soil miro-organisms were assessed using germination bioassay and antimicrobial assay, and allelochemicals were identified. In a germination bioassay, extract of cork tree inhibited at high concentration on germination of several crop seeds such as cabbage, lettuce, and cucumber. However, aqueous extracts inhibited powerfully growth of test organisms such as Streptococus aureus, S. aureus, S. typhimurium, and E. coli as bacteria, and Candida albicans as yeast, and Botrytis cineria and Alternata alternaria as fungi.. The cork tree extract showed strong antimicrobial activities against isolated soil fungi. The allelochemicals were separated using Silica gel, Sephadex LH-20 gel column chromatography and HPLC. The substances were analyzed by UV spectrometry and EI-mass spectrometry. The active allelochemicals were identified as isoquinoline alkaloids, berberine and palmatine.

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Comparison of Short-Term Toxicity Tests Based on Feeding Behavior and Temperature Control by Ceriodaphnia dubia (Ceriodaphnia dubia의 먹이섭생 기작과 온도조절에 근거한 급성독성조사법의 비교)

  • Park, Jong-Ho;Lee, Sang-Ill;Cho, Young-Oak
    • Journal of Korean Society on Water Environment
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    • v.20 no.1
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    • pp.48-54
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    • 2004
  • Two methods, a Ceriodaphnia algal uptake suppression test (CAUST) and a new toxicity test based on temperature control (TTBTC) which are based on feeding behaviour and temperature control, respectively, were developed and compared for the adoption as the better methodology for short-term toxicity screening. As previously published by Lee et aI., (1997), the CAUST method is based on the feeding behaviour of C. dubia and requires as little as 1 hour of contact time between C. dubia neonates and toxicant. However, even though CAUST requires only 1 hour of contact time, this method still take many hours for the preparation and measurement. Before the test starts, neonate digestive tracts were cleared by feeding yeast to the daphnids, Neonates were then exposed to toxicant, followed by addition of Scenedesmus subspiatus into the bioassay vessels. Daphnids were examined under the bright-field microscope with the presence of algae (indicated by a green colored digestive tract) or the absence of algae. Uptake indicated no toxic effect, whereas, absence of uptake indicated toxic inhibition. Unlike CAUST, the newly developed method (TTBTC) is based on just temperature control for the toxicity test of C. dubia. Initially, neonates are exposed to toxicants while the temperature of water bath containing media increased to $35.5^{\circ}C$. After 1.25 hour of contact time, the number of the daphnids, either live (no toxic effect) or dead (toxic effect), is counted without the aid of any instrument. In both methods, median effective concentrations ($EC_{50}$ values) were computed based on the results over a range of dosed toxicant concentrations. It showed that TTBTC was as sensitive as the standard 48-hour acute bioassay and CAUST. TTBTC and CAUST were much more sensitive than the I-hour I.Q. test and 30-minute Microtox. This study indicates that TTBTC is an easier and more rapid toxicity test than the standard 48-hour acute bioassay and even CAUST.

Detection of estrogenic hormone 17β-estradiol in soil samples by a recombinant yeast bioassay and supercritical fluid extraction

  • Shim, Jae-Han;Kim, Mi-Ra;Topp, Edward;Choi, Jeong-Heui;Mamun, Iqbal Rouf
    • Korean Journal of Environmental Agriculture
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    • v.27 no.4
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    • pp.447-455
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    • 2008
  • Recombinant yeast estrogenicity (YES) assay was used as a bioanalytical tool in order to screen $17{\beta}$-estradiol in the soil samples collected from different sites of South Korea. Solvent extraction and supercritical fluid extraction (SFE) methods were compared for the extraction of the estradiol from the soils. Most high detection of the estradiol based on YES assay was observed in the soils extracted with methanol. Different types of estrogenic hormones including $17{\beta}$-estradiol were suggested to be possibly exiting in the soils, since the methanol extracts of the soils showed an estrogenic activity that was not observed in the hexane extracts of the soil. SFE extracts showed estrogenic activity in some of the samples but methanol extract showed best activity.

Toxicity Evaluation of Endocrine Disrupting Chemicals Using Human HepG2 Cell Line, Lumbricus rubellus and Saccharomyces cerevisiae (HepG2 인간 세포주, Lumbricus rubellus 및 Saccharomyces cerevisiae를 이용한 내분비교란물질의 독성평가)

  • Sohn, Ho-Yong;Kim, Hong-Ju;Kum, Eun-Joo;Cho, Min-Seop;Lee, Jung-Bok;Kim, Jong-Sik;Kwon, Gi-Seok
    • Journal of Life Science
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    • v.16 no.6
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    • pp.919-924
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    • 2006
  • Toxicity evaluation systems for various chemicals and their metabolites have been developed during last decades. In this study, the acute toxicity of endocrine disrupting chemicals, such as endosulfan, bisphenol A, vinclozolin, and 3,5-dichloroaniline, was evaluated using HepG2 cell line, Lumbricus rubellus and Saccharomyces cerevisiae, respectively. The extents of toxicity of the chemicals in different bioassay systems varied substantially, such as endosulfan>3,5-dichloroaniline> bisphenol A in HepG2 cell line system, endosulfan>bisphenol A>3,5-dichloro aniline in L. rubellus system, and 3,5-dichloroaniline>endosulfan>bisphenol A in S. cerevisiae system. Meanwhile, no cytotoxicity was observed by treatment of vinclozolin in the evaluation systems. Our results suggest that earthworm and yeast are useful to evaluate acute toxicity of endocrine disrupting chemicals, and direct comparison of toxicity data from different bioassay systems is unattainable. Based on our results, we propose that the bioassay system with earthworm or yeast, a rapid, simple and economic system, could be applied as pre-test for the toxicity evaluation using human cell line or animals.

Identification and Characterization of Entomopathogenic Fungi Isolated from Bemisia tabaci in Korea (담배가루이(Bemisia tabaci )에서 분리한 곤충병원성진균의 동정 및 병원성 검정)

  • Park, Hyun-Rho;Ryu, Young-Hyun;Yeon, Il-Kyen;Nam, Sung-Hee;Kim, Dong-Geun;Han, Myung-Sae
    • Korean journal of applied entomology
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    • v.53 no.1
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    • pp.27-34
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    • 2014
  • Entomopathogenic fungi were isolated from Bemisia tabaci in an Oriental melon field, and their growth characteristics, factors related to a natural outbreak, and infectivity against Bemisia tabaci, Tetranychus urticae, and Frankliniella intonsa were investigated. The isolates had erect conidiophores bearing whorls of 4-6 phialides with a swollen base where cylindrical conidia of $3.0-3.4{\mu}m$ were attached. The isolates were identified as Isaria fumosorosea on the basis of morphological characteristics and an ITS sequence with 99% similarity. I. fumosorosea IFs-08 grew well on Sabouraud dextrose agar+yeast extract medium(3.2 mm/day/$24^{\circ}C$); it grew better at $35^{\circ}C$ than at $15^{\circ}C$. The isolates of I. fumosorosea-IFs were highly infective and killed 93.9-96.7% B. tabaci, 84.9-92.0% T. urticae, and 81.5-84.4% F. intonsa in bioassay, whereas three isolates (Isaria tenuipes, Isaria farinosa, and Isaria fumosorosea) from KACC showed a low infectivity of 10-20%. To the best of our knowledge, this is the first report of I. fumosorosea isolated from B. tabaci in Korea.

Expression of a Functional Human Tumor Necrosis Factor-${\alpha}$ (hTNF-$\alpha$) in Yeast Saccharomyces cerevisiae

  • Park, Seung-Moon;Mo, Ae-Young;Jang, Yong-Suk;Lee, Jae-Hwa;Yang, Moon-Sik;Kim, Dae-Hyuk
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.4
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    • pp.292-296
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    • 2004
  • The recombinant soluble human tumor necrosis factor-alpha (hTNF-$\alpha$) was expressed in a yeast Saccharomyces cerevisiae and its cytotoxicity was evaluated. A cDNA encoding hTNF-$\alpha$ was placed under the control of two different promoters: a glyceraldehyde-3-phosphate dehydrogenase (GPD) promoter and a yeast hybrid ADH2-GPD promoter, consisting of alcohol dehydrogenase II (ADH2) and the GPD promoter. A Northern blot analysis revealed that, although variation in the expression level of hTNF-$\alpha$ existed among transformants, the higher expression was obtained with the GPD promoter. Expressed hTNF-$\alpha$ protein (rhTNF-$\alpha$) was successfully secreted into the culture medium, producing 2.5 mg per liter of culture filtrate, with no changes in cell growth. The bioassay for observing the cytotoxicity to the murine L929 fibroblast cell line, with serial dilution of rhTNF-$\alpha$, indicated that the secreted rhTNF-$\alpha$ was bioactive and its dose-response was improved eight to ten times over that of the E. coli-derived rhTNF-$\alpha$.