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Optimizing enzyme properties to enhance dihydroxyacetone production via methylglyoxal biosensor development
Kaibo Zhang1,2, Mengying Li2,3, Jinsheng Wang2,4
1School of Chemistry and Life Science, Changchun University of Technology, Changchun, 130012, Jilin, China.
Microbial Cell Factories
|May 25, 2024
Summary
This study developed a novel methylglyoxal biosensor to screen for improved dihydroxyacetone (DHA) production enzymes. The biosensor successfully identified HdpA enzyme mutants that significantly increased DHA yield, offering a new method for cosmetic ingredient optimization.
Area of Science:
- Biotechnology
- Enzyme Engineering
- Metabolic Engineering
Background:
- Dihydroxyacetone (DHA) is a key cosmetic ingredient produced via dihydroxyacetone phosphate dephosphorylation.
- The enzyme dihydroxyacetone phosphate dephosphorylase (HdpA) from Escherichia coli is crucial for DHA production.
- Limited understanding of HdpA hinders efficient industrial production and engineering efforts.
Purpose of the Study:
- To develop a high-throughput screening tool for identifying HdpA hypermutants.
- To engineer a biosensor for indirect detection and selection of enhanced DHA production.
Main Methods:
- Constructed a methylglyoxal biosensor using the FrmR regulator and a modified constitutive promoter.
- Engineered hybrid promoters with FrmR binding sites to enable inducible expression.
- Validated the biosensor's response to methylglyoxal and its correlation with DHA production.
- Applied flow cytometry for sorting DHA-producing strains based on biosensor signal.
Main Results:
- The methylglyoxal biosensor demonstrated a 40-fold increase in promoter activity upon methylglyoxal addition.
- Fluorescence intensity correlated with methylglyoxal concentrations (10-500 μM).
- The biosensor successfully distinguished DHA-producing from non-producing strains.
- Screening identified two HdpA mutants (267G>T and D110G/G151C) with 68% and 114% increased DHA production, respectively.
Conclusions:
- A novel methylglyoxal biosensor was developed for genetically encoded screening.
- This biosensor provides a robust platform for indirectly measuring DHA levels via methylglyoxal.
- The system enables efficient screening of HdpA hypermutants to enhance DHA production for industrial applications.
