Oligosaccharides containing galactosyl moieties belong to two main groups: raffinose family oligosaccharides (RFO, α-GOS) and lactose-type β-galactooligosaccharides (β-GOS), both well-known for their prebiotic effect. The present review investigates the vast amounts of recent research on the structures of GOS and their beneficial impact. It focuses on the molecular interactions between GOS and probiotics in vitro and in vivo, the enzymology of the processes, and the genetic prerequisites for the synthesis and degradation of GOS by probiotic bacteria. The preferences of probiotic strains belonging to the Bifidobacterium and Lactobacillus genera are elucidated to form and degrade GOS of a certain length, structure, and linkages between monomers. A brief overview of the industrial production of β-GOS by natural and recombinant strains included the methods and production efficiency evaluation.

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.

Bacillus licheniformis 24 (BL24) is an efficient, non-pathogenic producer of 2,3-butanediol (2,3-BD). However, during inulin fermentation, the strain produces large amounts of exopolysaccharides (EPS), which interfere with the process’ performance. The present study aims to investigate the effect that inactivation of the sacB gene, encoding levansucrase in BL24, has on 2,3-BD production efficiency. Knockout of the sacB gene was accomplished via insertional inactivation. The sacB-knockout variant formed 0.57 g/L EPS from sucrose and 0.7–0.8 g/L EPS from glucose and fructose, a 15- and 2.5-fold reduction relative to the wild type, respectively. Likewise, during batch fermentation with soluble inulin Frutafit^® CLR, the mutant BLΔsacB produced significantly less EPS than the wild type, allowing the maintenance of pH at values favoring 2,3-BD synthesis. At pH 6.50, BLΔsacB reached a record titer of 128.7 g/L 2,3-BD, with productivity of 1.65 g/L/h, and a yield of 85.8% of the theoretical maximum. The obtained concentration of 2,3-BD is two-fold higher compared to that of the wild type. Subsequent RT-qPCR assays confirmed a successful sacB knockout. Three of the genes involved in inulin hydrolysis (sacA, sacC, and fruA) maintained their expression levels compared to the wild type, while that of levB increased. Although total EPS accumulation could not be completely eliminated via sacB gene knockout alone, the overall reduction in EPS content has enabled the highest yield of 2,3-BD from inulin to date, a promising result for the industrial production from inulin-rich substrates.