Bacterial Tolerance to 1-Butanol and 2-Butanol: Quantitative Assessment and Transcriptomic Response

The unique fuel characteristics of butanol and the possibility of its microbial production make it one of the most desirable environmentally friendly substitutes for petroleum fuels. However, the highly toxic nature of 1-butanol to the bacterial strains makes it unprofitable for commercial production. By comparison, 2-butanol has similar fuel qualities, and despite the difficulties in its microbial synthesis, it holds promise because it may be less toxic. This paper is the first comprehensive study to compare bacterial tolerance to different butanol isomers by examining the growth of 31 bacterial strains under 1-butanol and 2-butanol stress conditions. The presented results reveal that all tested strains showed a higher tolerance to 2-butanol than to 1-butanol at each solvent concentration (1%, 2%, and 3% v/v). Moreover, with an increased solvent concentration, bacterial cells lost their resistance to 1-butanol more rapidly than to 2-butanol. A comparison of the transcriptome profiles of the reference strains Bacillus subtilis ATCC 168 and E. coli ATCC 25922 disclosed a specific response to butanol stress. Most notably, in the presence of 2-butanol E. coli ATCC 25922 showed a reduced expression of genes for chaperones, efflux pumps, and the flagellar apparatus, as well as an enhancement of membrane and electron transport. B. subtilis, with 2-butanol, did not perform emergency sporulation or escape, as some global transcriptional stress response regulators were downregulated. The overexpression of ribosomal RNAs, pyrimidine biosynthesis genes, and DNA- and RNA-binding proteins such as pcrA and tnpB was crucial in the response.