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To reduce the cost of bacterial cellulose (BC) production, the possibility of using acetone-butanol-ethanol (ABE) fermentation wastewater with high COD value (18 050 mg l−1) for BC production by Gluconacetobacter xylinus was evaluated. After 7 days of fermentation, the highest BC yield (1·34 g l−1) was obtained. The carbon sources including sugars (glucose and xylose), organic acids (acetic acid and butyric acid) and alcohol compounds (ethanol and butanol) were utilized by G. xylinus simultaneously during fermentation. Although the COD decrease ratio (about 14·7%) was low, the highest BC yield on COD consumption (56·2%, g g−1) was relatively high and the remaining wastewater could be used for further BC fermentation. Besides, the environment of ABE fermentation wastewater showed small influence on the BC structure by comparison with the BC products obtained in traditional HS medium using field emission scanning electron microscope (FE-SEM), Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD). Overall, ABE fermentation wastewater is one promising substrate for BC production.