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Nanocellulose made by 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO)-catalyzed oxidation, described as TEMPO-oxidized cellulose nanofibers (TOCNs), has a high density of negative charges on its surface. Its use in microbial fermentation systems is expected to benefit microbial process stability. In particular, microbial stability is strongly required in acetone-butanol-ethanol (ABE) fermentation associated with the solvent-extraction process of butanol production. Here, TOCNs derived from oil palm empty fruit bunches pulp were added to extractive ABE fermentation media containing glucose as a main source, which can be potentially obtained from biomass by saccharification. Then, microbial fermentation was carried out using free or immobilized bacterial cells, to produce butanol from glucose. The presence of TOCNs induced higher total butanol production in broth by improving the growth environment of Clostridium saccharoperbutylacetonicum N1-4, which was used as the butanolproducing strain. Microscopic analysis revealed that the spider-web-like TOCN network helped to entrap bacterial cells in alginate beads, by ionic crosslinking of TOCNs and alginates via Ca2+ ions, to increase stability of bacterial cells in the composite gel beads. The addition of TOCNs to fermentation media had significant positive effects on the total butanol yield. [ABSTRACT FROM AUTHOR]