Interfacial Engineering Circumvents Activity-Stability Trade-Off in L-Threonine Aldolase for High-Yield L-threo-MTPS
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View abstract on PubMed
Summary
This summary is machine-generated.Interfacial engineering enhanced L-Threonine aldolase (LTA) stability and activity, improving β-hydroxy-α-amino acids (β-HAAs) synthesis. This breakthrough advances industrial production of valuable amino acid derivatives.
Area Of Science
- Biocatalysis and Enzyme Engineering
- Protein Engineering
- Industrial Biotechnology
Background
- L-Threonine aldolase (LTA) is crucial for synthesizing β-hydroxy-α-amino acids (β-HAAs).
- Existing LTA variants suffer from limited thermostability and an activity-stability trade-off, hindering industrial applications.
- Targeting nonconserved residues at subunit interfaces offers a potential strategy for enzyme improvement.
Purpose Of The Study
- To enhance the thermostability and catalytic activity of *Faecalimicrobium dakarense* LTA through interfacial engineering.
- To investigate the impact of mutations at subunit interfaces on enzyme performance and stability.
- To optimize the production of agrochemical precursors using engineered LTA.
Main Methods
- Site-directed mutagenesis was employed to introduce specific amino acid substitutions (L17F/V129R/L206W) at subunit interfaces.
- Enzyme activity, thermostability (T50^60, Tm), and product yield were assessed under varying conditions.
- Molecular dynamics simulations were used to elucidate the structural basis for enhanced enzyme performance.
Main Results
- The M3 variant displayed a 1.23-fold increase in specific activity and significantly improved thermostability (+20.5 °C for T50^60, +13.1 °C for Tm).
- M3 achieved a 2.49-fold higher yield of the agrochemical precursor L-threo-4-methylsulfonyl-phenylserine in a 1 L system.
- Scale-up to 10 L demonstrated high conversion (95.7%), diastereomeric excess (84.9% de), and space-time yield (10.5 g·L⁻¹·h⁻¹).
Conclusions
- Interfacial engineering effectively decoupled the activity-stability trade-off in LTA, leading to a superior biocatalyst.
- The M3 variant demonstrates significant potential for the industrial synthesis of β-HAAs and related compounds.
- This strategy provides a generalizable approach for improving multimeric enzymes for biotechnological applications.
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