Inhibition effect of isopropanol on acetyl-CoA synthetase expression level of acetoclastic methanogen, Methanosaeta concilii

Isopropanol is a widely found solvent in industrial wastewaters, which have commonly been treated using anaerobic systems. In this study, inhibitory effect of isopropanol on the key microbial group in anaerobic bioreactors, acetoclastic methanogens, was investigated. Anaerobic sludges in serum bottles were repeatedly fed with acetate and isopropanol; and quantitative real-time PCR was used for determining effect of isopropanol on the expression level of a key enzyme in acetoclastic methane production, acetyl-CoA synthetase of Methanosaeta concilii. Active Methanosaeta spp. cells were also quantified using Fluorescent in situ hybridization (FISH). Transcript abundance of acetyl-CoA synthetase was 1.23±0.62×10 ⁶mRNAs/mL in the uninhibited reactors with 222mL cumulative methane production. First exposure to isopropanol resulted in 71.2%, 84.7%, 89.2% and 94.6% decrease in mRNA level and 35.0%, 65.0%, 91.5% and 100.0% reduction in methane production for isopropanol concentrations of 0.1M, 0.5M, 1.0M and 2.0M, respectively. Repeated exposures resulted in higher inhibitions; and at the end of test, fluorescent intensities of active Methanosaeta cells were significantly decreased due to isopropanol. The overall results indicated that isopropanol has an inhibitory effect on acetoclastic methanogenesis; and the inhibition can be detected by monitoring level of acetyl-CoA transcripts and rRNA level.