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Updated: Jun 19, 2026

Enzymatic Cascade Reactions for the Synthesis of Chiral Amino Alcohols from L-lysine

Enzymatic Cascade Reactions for the Synthesis of Chiral Amino Alcohols from L-lysine

Published on: February 16, 2018

Engineering a PLP-dependent Mannichase.

Shaonan Liu¹, Yang Hai¹

¹Department of Chemistry and Biochemistry, University of California, Santa Barbara, Santa Barbara, CA, United States.

Methods in Enzymology

|October 30, 2025

Summary

This summary is machine-generated.

Researchers engineered a pyridoxal 5'-phosphate (PLP)-dependent enzyme, LolT, into a Mannichase. This biocatalyst enables efficient, enantioselective synthesis of α,β-diamino acids, expanding synthetic chemistry applications.

Keywords:

Biocatalysis LolT Mannich reaction PLP-dependent enzyme Protein engineering α,β-Diamino acid

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Last Updated: Jun 19, 2026

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Area of Science:

Biocatalysis and Enzyme Engineering
Synthetic Organic Chemistry
Medicinal Chemistry

Background:

Pyridoxal 5'-phosphate (PLP)-dependent enzymes are known for diverse bond-forming capabilities.
A PLP-dependent enzyme for intermolecular Mannich reactions yielding α,β-diamino acids was previously undescribed.
LolT, a PLP-dependent Mannich cyclase, was investigated for repurposing.

Purpose of the Study:

To engineer LolT into a synthetically valuable Mannichase for α,β-diamino acid synthesis.
To enhance the non-native Mannichase activity of LolT through protein engineering.
To establish a method for repurposing PLP enzymes for novel transformations.

Main Methods:

Iterative site-saturation mutagenesis of LolT.
Development of a double high-throughput screening platform.
Biochemical and analytical validation of enzyme variants.

Main Results:

Engineered LolT variant (LolT^V4) showed >60-fold improvement in catalytic turnover.
Enabled gram-scale, one-step, enantioselective synthesis of L-tambroline.
Demonstrated successful repurposing of a PLP enzyme for a non-natural reaction.

Conclusions:

Engineered LolT serves as a valuable Mannichase for synthesizing α,β-diamino acids.
This work provides a blueprint for expanding biocatalysis scope using PLP enzymes.
The developed methods facilitate the discovery of novel biocatalysts for synthetic and medicinal chemistry.