Engineered bakers yeast as a sensitive bioassay indicator organism for the trichothecene toxin deoxynivalenol.
The aim of this study was to increase the sensitivity of Saccharomyces cerevisiae towards trichothecene toxins, in particular to deoxynivalenol (DON), in order to improve the utility of this yeast as a bioassay indicator organism. We report the construction of a strain with inactivated genes (PDR5, PDR10, PDR15) encoding ABC transporter proteins with specificity for the trichothecene deoxynivalenol, with inactivated AYT1 (encoding a trichothecene-3-O-acetyltransferase), and inactivated UBI4 and UBP6 genes. Inactivation of the stress inducible polyubiquitin gene UBI4 or the ubiquitin protease UBP6 increased DON sensitivity, the inactivation of both genes had a synergistic effect. The resulting pdr5 pdr10 pdr15 ayt1 ubp6 ubi4 mutant strain showed 50% growth inhibition at a DON concentration of 5 mg/l under optimal conditions. The development of a simple two step assay for microbial DON degradation in 96 well microtiter format and its testing with the DON detoxifying bacterium BBSH 797 is reported.
Abolmaali
Shamsozoha
S
University of Natural Resources and Applied Life Sciences, Muthgasse 18, A-1190 Vienna, Austria.
Mitterbauer
Rudolf
R
Spadiut
Oliver
O
Peruci
Michaela
M
Weindorfer
Hanna
H
Lucyshyn
Doris
D
Ellersdorfer
Günther
G
Lemmens
Marc
M
Moll
Wulf-Dieter
WD
Adam
Gerhard
G
D018528:ATP-Binding Cassette Transporters / Q000235:genetics; D000123:Acetyltransferases / Q000235:genetics; D001419:Bacteria / Q000378:metabolism; D010450:Endopeptidases / Q000235:genetics; D017353:Gene Deletion; D020128:Inhibitory Concentration 50; D008828:Microbiological Techniques; D012441:Saccharomyces cerevisiae / Q000187:drug effects* / Q000235:genetics / Q000254:growth & development; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics; D012680:Sensitivity and Specificity; D014255:Trichothecenes / Q000378:metabolism / Q000633:toxicity*; D025822:Ubiquitin C / Q000235:genetics
D018528:ATP-Binding Cassette Transporters; C529225:PDR10 protein, S cerevisiae; C529226:PDR15 protein, S cerevisiae; C092438:PDR5 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D014255:Trichothecenes; C417885:UBI4 protein, S cerevisiae; D025822:Ubiquitin C; D000123:Acetyltransferases; C110205:trichothecene 3-O-acetyltransferase; D010450:Endopeptidases; C109028:UBP6 protein, S cerevisiae; C007262:deoxynivalenol