DISCUSSION

The experiment of inoculum preparation for bacterial cellulose by shake flask

fermentation were aimed to study the effect of pH on the medium, to study the reducing sugar
of the medium and to study the production of bacterial cellulose using both agitated and static
condition.

In this experiment, one of the important parameters that might affects the production of
the bacterial cellulose is pH. Basically from the previous study by Rangaswamy, Vanitha, &
Hungund, (2015) stated that pH plays an important role in cellulose production and the
optimum pH for the cellulose production is 5.4 to 6.4. Moreover, the optimum pH for the
Acetobacter xylinum to growth is 5 to 6 and that is why in this experiment the medium and the
inoculum medium pH were adjust to 6.4 at the beginning. From the experiment, 4 sample has
been done at static condition and 4 sample has been done at agitated condition. Figure 1 is the
graph of pH for static condition. The graph trend in the Figure 1 represents the pH of the
medium shows that most of the sample either at static or agitated condition decreased their pH
value started from day 3. Same goes with graph in Figure 2 that represents the agitated
condition, the trendline of the graph also shows the decreasing of the pH value to acid. This is
because the Acetobacter xylinum produce acetic acid during their growth and the acetic acid
was reported as the by-product during the synthesis of bacterial cellulose (Zahan, Pa’e, &
Muhamad, 2015).

Other than that, the reducing sugar also has been observe in this experiment by refer to
the standard curve of glucose in Figure 5. The Figure 3 represents the graph of the results
reducing sugar for the static condition whereas the Figure 4 represents the graph of the results
reducing sugar for agitated condition. From the trendline of the graph in Figure 3 and 4, the
concentration of reducing sugar for both conditions were decreased as the fermentation day
increase. This is because during the bacterial cellulose production, the glucose act as the energy
source for the Acetobacter xylinum. So, the decrease of the glucose concentration is because
the Acetobacter xylinum consume. But, from the previous study by Zahan, Pa’e, Seng, &
Muhamad, (2013) stated that if the medium is at the high of glucose concentration, the
excessive glucose was actively converted by the membrane-bound Acetobacter
dehydrogenases into glutamic acid. This high glutamic acid presence in the medium will
decrease the yield of the production compared to the low glutamic acid. The process to produce
bacterial cellulose will be more efficient for in a medium with glucose (Masaoka, Ohe, &
Sakota, 1993).

Thus, the bacterial cellulose weight also has been observed for all sample but not all of
the sample contain bacterial cellulose only sample static 2, 3, 4 and agitated 1, 2, 4 contain
bacterial cellulose as shown in Figure 7 above. Although all of the sample is sample, but the
production of the bacterial cellulose is different. This is because the inoculum that has been
used for each of the sample is from the different source. Some sample might get the inoculum
that contain more cultured of Acetobacter xylinum which can contribute to the high production
of bacterial cellulose. Based on the observation from the results at Figure 6, the agitated
condition tends to produce more bacterial cellulose compared to the static condition. This is
because at the agitated conditions, the Acetobacter xylinum could produce more bacterial
cellulose with the condition where the flask is efficient for oxygen uptake and homogenous
environment for the bacterial to growth (Zahan et al., 2013).

Figure 7: Results for bacterial cellulose before dry.

REFERENCES

Masaoka, S., Ohe, T., & Sakota, N. (1993). Production of cellulose from glucose by
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Rangaswamy, B. E., Vanitha, K. P., & Hungund, B. S. (2015). Microbial Cellulose

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