A syngas compositional database with focus on trace impurities was established. For this work, ammonia (NH3) and benzene (C6H6) effects on cell growth, enzymatic activities of hydrogenase and alcohol dehydrogenase (ADH), and product formation were studied. NH3, after entering media, will be converted rapidly to NH4+, which will raise the total osmolarity of the media. NH3, as a common nutrient for the cell growth, is not the real culprit for cell growth inhibition. In essence, it is the high osmolarity resulting from the accumulation of NH4+ in the media which disrupts the normal regulation of the cells. It was concluded that at NH4+ concentration above 250 mM, the cell growth was substantially inhibited. However, P11 cells used in this study can likely adapt to an elevated osmolarity (up to 500 mM) although the mechanism is unknown. It was also found that higher osmolarity will eventually lead to higher ethanol per cell density. In conclusion, NH3 needs to be cleaned out of syngas feeding system. The realistic C6H6 concentration in the media coming from a gasifier was simulated in bioreactors and was measured by a GC/MS. The most realistic C6H6 concentration in the media was around 0.41 mM (upper limit 0.83 mM). However, five elevated concentrations of 0.64, 1.18, 1.72, 2.33, and 3.44 mM were doped into the media. It was found that at 3.44 mM cell growth and ethanol production were significantly affected. However, there was only negligible adverse effect on cell growth and ethanol production at 0.41 mM, which is the expected concentration in bioreactors exposed to syngas. Therefore, it is unnecessary to remove C6H6 from the gas feeding stream. A kinetic model for hydrogenase activity that included inhibition effects of NH4+ and C6H6 was developed. Experimental results showed that NH4+ is a non-competitive inhibitor for hydrogenase activity with KNH4+ of (649 ± 35) mM and KH2 of (0.19 ± 0.1) mM. This KH2 value is consistent with those reported in literature. C6H6 is also a non-competitive inhibitor but a more potent one compared to NH4+ (KC6H6=11.4 ± 1.32 mM). A KH2 value of (0.196 ± 0.022) mM is also comparable with literature and also with the NH4+ study. At a realistic C6H6 concentration of 0.41 mM expected in bioreactors exposed to syngas, hydrogenase activity is expected to be reduced by less than 5%. Forward ADH activity was not adversely affected up to 200 mM [NH4+].From the current work, NH3 should be targeted for removal but it is not necessary to remove C6H6 when designing an efficient gas cleanup system.



College and Department

Ira A. Fulton College of Engineering and Technology; Chemical Engineering



Date Submitted


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syngas impurity, hydrogenase activity, ethanol formation, inhibition, fermentation