Summary: | Lignocellulosic substrates are considered to be crucial substrates for the production of biofuels. The main objective of the study is to attempt to produce bioethanol using bio-wastes such as mango peels, orange peels, and tapioca shells as renewable sources by employing three bacteria viz., <i>Enterobacter cloacae</i> (ICBP1), <i>Pseudomonas aeruginosa</i> (ICBP7), and <i>Bacillus cereus</i> (ICBP15), which were chosen to produce cellulase enzymes using the submerged fermentation method, which is a novel method for the production of bioethanol. The “zone of clearance” in bacterial growth on CMC agar plates determined the choice. The mixed culture infected units produced a more reduced sugar, i.e., the presence of aldehyde and ketones except sucrose. At 72 h, greater than 41.0 ± 0.48 mL and 0.83 ± 0.07% of ethanol was recovered. This contrasts with the reduced quantities at 24 and 48 h. SDS-PAGE examination showed that the three cellulose-producing bacterial strains (ICPB1, ICPB7, and ICPB15) had enzyme molecular weights of 80–100, 20–30, and 14–20 kDa, respectively, compared to the other 17 isolates. Fourier-transform infrared (FTIR) spectroscopy was used to estimate the bioethanol. The spectrum bands from 1700 to 1800 cm<sup>−1</sup> showed bioethanol’s unique absorption characteristics, and GC-MS confirmed 31.38% ethanol. The findings of the research demonstrate that the utilization of fermentation technology, specifically employing microbes, to produce bioethanol from bio-wastes such as fruits and vegetables has the potential to address the worldwide fuel energy requirements.
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