Engineered Microbial Consortium for the Efficient Conversion of Biomass to Biofuels Page: 75
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From the above bar plots, it is clear that the wild type ZM4 does not make ethanol from
cellulose as earlier suggested. The ethanol content quantified from the first two experiments were
carryovers from the inoculum and with the washed cells, this was no longer an issue. Traces of
ethanol production continued to be seen with the wild type ZM4 alone for the five days and the
ten days samples even after the thorough washing. From the above plots, the ethanol detected from
wild type ZM4 alone after washing did not exceed 0.2 mM.
ZM4 is a very efficient ethanologen and the background noise seen here is likely as a result
of letting the cells grow to an OD 2=600 above 1.50 in the RM medium used for the inoculation
of the media containing cellulose. Though the cells were washed before use, it is likely that there
was still enough residual ethanol that was carried over to be detected in these assays. One clear
observation is the significant reduction in the amount of ethanol observed from wild type ZM4
cultures alone, once determined at 2.52 mM at its peak but less than 0.2 mM from the last trials
that involved washing the cells before use. The five-day cultures of batches 1, 2 and 3 suggest a
synergy in the co-culture, with batch 2 producing the highest amount of ethanol at slightly below
0.4 mM concentration; far less than was observed when the unwashed wild type ZM4 inoculum
was used. From the 10 days cultures of the same batches, batch 1 cultures showed improved
ethanol concentration across for Clostridium alone and for the co-culture of wild type ZM4 +
Clostridium. Batch 2 however had no ethanol detected for the co-culture, suggesting that the
Clostridium could have gone into the dormant/sporulation stage and the wild type ZM4 in the
medium could be all dead. A similar but different pattern can be seen in batch 3, where no ethanol
activity can be seen in the monocultures but still observable in the co-culture, again suggesting a
synergy.75
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Anieto, Ugochukwu Obiakornobi. Engineered Microbial Consortium for the Efficient Conversion of Biomass to Biofuels, dissertation, August 2014; Denton, Texas. (https://digital.library.unt.edu/ark:/67531/metadc699973/m1/86/: accessed July 17, 2024), University of North Texas Libraries, UNT Digital Library, https://digital.library.unt.edu; .