A Sensitive and Inexpensive Yeast Bioassay for the Mycotoxin Zearalenone and Other Compounds with Estrogenic Activity

Mitterbauer, Rudolf; Weindorfer, Hanna; Safaie, Naser; Krska, Rudolf; Lemmens, Marc; Ruckenbauer, P.; Kuchler, Karl; Adam, Gerhard

doi:10.1128/aem.69.2.805-811.2003

Cited by 43 publications

(38 citation statements)

References 48 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…We previous reported that the PDR5 mutant strain shows high susceptibility to both DON and T-2 toxin [13], suggesting that the PDR5 coding protein strongly influences the elimination of trichothecene mycotoxin. Several similar results have already been reported [29] [30]. Thus the growth inhibition and PDR5 repression found in the present study may be of relevance to DON toxicity.…”

Section: Discussionsupporting

confidence: 77%

Gene expression profile of MAP kinase PTC1 mutant exposed to deoxynivalenol

忠宏

由美子

2011

CBIJ

View full text Add to dashboard Cite

Deoxynivalenol (DON) is a secondary metabolite that is generated by Fusarium species, which seriously affects both humans and livestock. Protein synthesis inhibition and ribotoxic stress, caused by induction of the mitogen activated protein kinase (MAPK) cascade, are thought to be responsible for the majority of DON toxicity. However, as DNA damage has also been reported, it is necessary to clarify all sources of toxicity. In this study, we conducted a DON exposure test using the PTC1 yeast mutant with disrupted MAPK-related genes, and observed gene expression changes using DNA microarray analysis. Our results indicated changes in the expression of genes associated with protein synthesis inhibition, as well as with DNA damage. At the same time, genes related to the synthesis of folic acid, a coenzyme in DNA synthesis, were inhibited. To complement the dysfunction of these genes, the growth media was supplemented with folic acid. As a result, the recovery of growth was confirmed, although it was a consistent effect and it did not reflect differences in susceptibility to DON toxicity.

show abstract

Section: Discussionsupporting

confidence: 77%

Gene expression profile of MAP kinase PTC1 mutant exposed to deoxynivalenol

忠宏

由美子

2011

CBIJ

View full text Add to dashboard Cite

show abstract

“…Mycotoxicity has also been thoroughly examined in yeast. Mutants with two or more gene deletions that disrupt the pleiotropic drug resistance 5 (PDR5) gene, which encodes a multidrug resistance protein, have lead to increases in mycotoxin susceptibility (McCormick et al, 1999, Mitterbauer et al, 2003, indicating that other stress response genes might also influence mycotoxin resistance in yeast. Recently, studies on mycotoxin-yeast interactions have revealed that metabolism and stress response genes are induced under patulin (Iwahashi et al, 2006) and citrinin (Iwahashi et al, 2007) exposure.…”

Section: Introductionmentioning

confidence: 99%

Mycotoxin Toxicity in Saccharomyces cerevisiae Differs Depending on Gene Mutations

Suzuki

Sirisattha

Mori

et al. 2009

FSTR

View full text Add to dashboard Cite

Mycotoxin inhibits the growth of Saccharomyces cerevisiae mutants with disrupted stress response genes, but no that of wild type yeast even at high concentrations. The S. cerevisiae genome database (SGD) project has recently generated many gene disruptants, including those with disrupted stress responseassociated genes. The present study examined the relationship between mycotoxicity and the disruption of functional yeast genes, especially those involved with stress responses. Growth tests showed a major impact in strains other than those with disrupted stress response genes as well as mycotoxin susceptibility of several strains by sodium dodecyl sulfate.

show abstract

“…Although a low-cost tool for monitoring ZEN levels with an engineered yeast strain has been developed, this method gives false-positive results because of other interfering estrogenic compounds, and it takes a long time to obtain results (18). In contrast, detoxification of mycotoxins appears to be a more attractive approach.…”

mentioning

confidence: 99%

Metabolism of Zearalenone by Genetically Modified Organisms Expressing the Detoxification Gene from Clonostachys rosea

Takahashi-Ando¹,

Ohsato²,

Shibata

et al. 2004

Appl Environ Microbiol

104

View full text Add to dashboard Cite

Zearalenone (ZEN) is converted to a nontoxic product by a lactonohydololase encoded by zhd101. An enhanced green fluorescent protein (EGFP) gene was fused to zhd101 (i.e., egfp::zhd101) and expressed in Escherichia coli. Both recombinant ZHD101 and EGFP::ZHD101 were purified to homogeneity and characterized. Maximal activity of ZHD101 toward ZEN was measured at approximately 37 to 45°C and pH 10.5 (k cat at 30°C, 0.51 s ؊1 ). The enzyme was irreversibly inactivated at pH values below 4.5 or by treatment with serine protease inhibitors. ZHD101 was also active against five ZEN cognates, although the efficiencies were generally low; e.g., the k cat was highest with zearalanone (1.5 s ؊1 ) and lowest with ␤-zearalenol (0.075 s ؊1 ). EGFP:: ZHD101 had properties similar to those of the individual proteins with regard to the EGFP fluorescence and lactonohydrolase activity. Fortuitously, EGFP::ZHD101 exhibited a good correlation between the fluorescence intensity and reaction velocity under various pH conditions. We therefore used egfp::zhd101 to visually monitor the lactonohydrolase activity in genetically modified organisms and evaluated the usefulness of zhd101 for in vivo detoxification of ZEN. While recombinant E. coli and transgenic rice calluses exhibited strong EGFP fluorescence and completely degraded ZEN in liquid media, recombinant Saccharomyces cerevisiae gave poor fluorescence and did not eliminate all the toxicity of the mycotoxin in the medium; i.e., the rest of ZEN was transformed into an unfavorable substrate, ␤-zearalenol, by an as-yet-unidentified reductase and remained in the medium. Even so, as much as 75% of ZEN was detoxified by the yeast transformant, which is better than the detoxification system in which food-grade Lactobacillus strains are used (H. El-Nezami, N. Polychronaki, S. Salminen, and H. Mykkuäne, Appl. Environ. Microbiol. 68: [3545][3546][3547][3548][3549] 2002). An appropriate combination of a candidate host microbe and the codon-optimized synthetic gene may contribute significantly to establishing a mycotoxin detoxification system for food and feed.Zearalenone (ZEN), 6-(10-hydroxy-6-oxo-trans-1-undecenyl)-␤-resorcylic acid lactone (Fig. 1), is a nonsteroid estrogenic mycotoxin that is produced by numerous Fusarium species in pre-or postharvest cereals (22). Under environmental conditions favorable for fungal growth, high levels of ZEN are frequently found in maize and other small grains, such as wheat and barley. The keto group at C-6Ј is reduced to a more estrogenic compound, ␣-zearalenol, and/or its stereoisomeric cognate ␤-zearalenol (which has less estrogenicity than ZEN) by various microorganisms, including industrial yeast strains (4) and animal intestinal microbes (13). These mycotoxins have adverse health effects on various animals that ingest moldinfected cereals and cereal-derived food products (16).ZEN causes severe morphological and functional disorders of reproductive organs in livestock, especially female swine (7). In feeding experiments with female swine, clear clin...

show abstract

A Sensitive and Inexpensive Yeast Bioassay for the Mycotoxin Zearalenone and Other Compounds with Estrogenic Activity

Cited by 43 publications

References 48 publications

Gene expression profile of MAP kinase PTC1 mutant exposed to deoxynivalenol

Gene expression profile of MAP kinase PTC1 mutant exposed to deoxynivalenol

Mycotoxin Toxicity in Saccharomyces cerevisiae Differs Depending on Gene Mutations

Metabolism of Zearalenone by Genetically Modified Organisms Expressing the Detoxification Gene from Clonostachys rosea

Contact Info

Product

Resources

About