Symposiumgids 2008 - Bodembreed

13C/12C - MtBE
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A
R 2 = 0,98
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MTBE degraded (%)
13C/12C - MtBE
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B
Figure 2 Stable isotope shifts correlate linearly with amount of MtBE degraded (A) and TBA
produced (B).
Finally, we have developed molecular detection methods to detect MtBE degrading organisms. We
evaluated the commonly used 16S rDNA TaqMan assay for quantitative detection of the described
MtBE-degrader Methylibium PM1 and found that it was unspecific, producing essentially false positive
results.
We have developed two sets of primer/probe combinations for quantitative realtime PCR detection of
functional genes involved in MtBE degradation. One set is targetting the putative MtBEmonooxygenase
of strain PM1 (Methylibium), and the other set is targetting the iso-butyryl-CoA
mutase of Aquincola. This latter enzyme was recently described to be essential for down-stream
degradation of MTBE and found in the genome of various MTBE-degraders. We have prepared
calibration curves and evaluated Q-PCR assays and found that they indeed amplify the targeted
sequences quantitatively. These assays are currently validated on various MtBE and/or TBA
degrading cultures.
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R 2 = 0,98
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TBA produced (µg/L)