Engineering an AlkS-PalkB Transcription Factor-Based Biosensor With Improved Sensitivity to Isobutanol and Other Short-Chain Alcohols.

ABSTRACT: Isobutanol, a promising biofuel with higher energy content than ethanol, presents a sustainable alternative through biosynthesis. However, enhancing yield remains challenging due to the inefficiencies in microbial synthesis. This study introduces a transcription factor-based biosensor using the AlkS-PalkB system in Escherichia coli, which correlates green fluorescence with isobutanol concentration. Employing directed evolution, we modified AlkS to detect isobutanol, significantly improving biosensor specificity. Initial modifications increased the dynamic response from non-detectable to a 2.60-fold change. Subsequent optimisations through site-directed mutagenesis and promoter engineering further enhanced this response to a 5.56-fold change, equivalent to a 114% increase. Although engineer

SUBMITTER: Chen J 

PROVIDER: S-EPMC12740625 | biostudies-literature | 2025 Dec

REPOSITORIES: biostudies-literature