Search for: All records

Abstract Adaptive laboratory evolution (ALE) can be used to make bacteria less susceptible to oxidative stress. An alternative to large batch scale ALE cultures is to use microfluidic platforms, which are often more economical and more efficient. Microfluidic ALE platforms have shown promise, but many have suffered from subpar cell passaging mechanisms and poor spatial definition. A new approach is presented using a microfluidic Evolution on a Chip (EVoc) design which progressively drives microbial cells from areas of lower H₂O₂concentration to areas of higher concentration. Prolonged exposure, up to 72 h, revealed the survival of adaptive strains ofLacticaseibacillus rhamnosusGG, a beneficial probiotic often included in food products. After performing ALE on this microfluidic platform, the bacteria persisted under high H₂O₂concentrations in repeated trials. After two progressive exposures, the ability ofL. rhamnosusto grow in the presence of H₂O₂increased from 1 mmH₂O₂after a lag time of 31 h to 1 mmafter 21 h, 2 mmafter 28 h, and 3 mmafter 42 h. The adaptive strains have different morphology, and gene expression compared to wild type, and genome sequencing revealed a potentially meaningful single nucleotide mutation in the protein omega‐amidase.