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Comparative assessment of acid and enzyme pretreatment of Spirodella polyrhiza for bioethanol production

By: Asma Khattak, Zulfiqar Ali Malik, Abdul Haq, Qurrat ul ain Rana, Haji Khan, Fazl ur Rehman, Samiullah Khan, Aamer Ali Shah, Malik Badshah

Key Words: Amylase, Bacteria, Reducing sugar, Yeast fermentation, Bioethanol.

Int. J. Biosci. 18(6), 103-119, June 2021.


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Duckweeds are considered more sustainable compared to conventional crops due to no feed versus fuel competition. However, the feedstocks used for bioethanol production require an efficient pretreatment to increase the sugar yield resulting in reduced cost of the process. To combat low sugar yield challenges, five isolates named AG 1, AG 2, AG 3, AG 4 and AG 5 were isolated from soil. The bacterial isolate AG 3 (Bacillus AS1 MK321577) showed the highest amylase activity with the DNS method. The Bacillus AS1 MK321577 strain was evaluated for saccharification of Spirodella polyrhiza followed by bioethanol production via Saccharomyces cerevisiae. The specific activity of amylase was found to be 0.396 U/mg/min after 24 h incubation. Maximum amylase activity was observed in boiled potato peels as carbon sources in the production media at pH 6 and 45 ºC. Placket-Burman’s design for optimization of physical and nutritional parameters was used. The parameters selected for optimization were KH2PO4, pH, inocula, CaCl2, MgCl2, temperature, starch, MgSO4.7H2O, reaction time, NaCl and NaNO3. The current study reports that Bacillus AS1 MK321577 strain has the highest amylase activity. Acid pretreatment of S. polyrhiza was also carried out and compared with the amylase pretreatment. Overall, the highest reducing sugars yield (0.432 mg/ml) and highest bioethanol production (99%) were observed in the amylase pretreated sample. In comparison to acidic pretreatment, enzymatic pretreatment is a more environmentally friendly process and resulted in a 15% enhanced yield of reducing sugars.

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Comparative assessment of acid and enzyme pretreatment of Spirodella polyrhiza for bioethanol production

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Asma Khattak, Zulfiqar Ali Malik, Abdul Haq, Qurrat ul ain Rana, Haji Khan, Fazl ur Rehman, Samiullah Khan, Aamer Ali Shah, Malik Badshah.
Comparative assessment of acid and enzyme pretreatment of Spirodella polyrhiza for bioethanol production.
Int. J. Biosci. 18(6), 103-119, June 2021.
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