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The pH of the mixture of microalgal biomass and inoculum (fermenting microbes) ranged from 5.7 to 6.8 at the beginning of fermentation, and the final pH values after the 23 days of fermentation were nearly identical (5.5 ± 0.2) in all sets of tested samples.
This problem was partially solved by sparging air containing 12% oxygen at the beginning of fermentation.
Besides, 150 μL antifoam (Sigma, USA) was added at the beginning of fermentation to control the formation of foam.
Figure 1 shows the laccase activity, which increased from the beginning of fermentation in BMF, with maximal activity observed at 408 h (239 U/mL).
It can be seen that, compared to the beginning of fermentation, there is an increase in the fractions of 4.62 and 12.41% after 48 and 72 h, respectively.
When adding l-alanine and d-tryptophan (5 and 10 mM) at the beginning of fermentation, the CHA accumulation declined, as shown in Fig. 3a.
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Biomass was immobilised in situ by natural adsorption, i.e. through direct contact between cells for the inoculum and carrier particles at the beginning of fermentations.
The lowest bacterial inoculation level (10cells/mL) simulates typical contamination concentrations found in industry at the beginning of fermentations [ 12– 14], whereas the highest bacterial inoculation level exemplifies an extreme case of contamination.
Very-high gravity fermentation causes high stress, in particular osmostress in the beginning of the fermentation and ethanol stress at the end of the fermentation.
The increased concentration of fermentable sugars at the beginning of the fermentation phase in SSF following prehydrolysis did not affect the overall ethanol yield in the present study.
The observed different trend could be related to a lower pH value measured in C1 fermentation, which decreased from 5.1 at the beginning of the fermentation to 3.7 after two refreshment steps; it could contribute to control the growth of non-acidophilic bacteria.
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