Engineering a PLP-dependent Mannichase.

Shaonan Liu, Yang Hai

Abstract read

In one paragraph

Article in Methods in enzymology, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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1 · What the graph read from it

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2 · The registry

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3 · Its place in the literature

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0 citing papers in PubMed.

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4 · The record

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5 · Who and what money

Authors and funding

2 authors.

Shaonan LiuDepartment of Chemistry and Biochemistry, University of California, Santa Barbara, Santa Barbara, CA, United States.

Yang HaiDepartment of Chemistry and Biochemistry, University of California, Santa Barbara, Santa Barbara, CA, United States. Electronic address: hai@chem.ucsb.edu.

Funding

Understanding the specificity and non-native catalytic activity of PLP-dependent enzymesR35GM151205 · NIGMS · UNIVERSITY OF CALIFORNIA SANTA BARBARA · PI Yang Hai · 2023 to 2026

$1.8M

NIGMS NIH HHS R35 GM151205

6 · The paper itself

Abstract

Pyridoxal 5'-phosphate (PLP)-dependent enzymes are versatile biocatalysts known for their ability to form diverse C-C, C-N, and C-S bonds. Despite this catalytic diversity, a PLP-dependent enzyme capable of promoting an intermolecular Mannich reaction to access α,β-diamino acids has not been described. Here, we report the engineering of LolT, a PLP-dependent Mannich cyclase from the loline alkaloid biosynthetic pathway, into a synthetically valuable Mannichase. Using iterative site-saturation mutagenesis and a double high-throughput screening platform, we identified mutations that significantly enhance LolT's non-native Mannichase activity. The best-performing variant, LolT

Indexed as

Protein EngineeringPyridoxal PhosphateBiocatalysisHigh-Throughput Screening AssaysMutagenesis, Site-DirectedMutationPyridoxal PhosphateBiocatalysisLolTMannich reactionPLP-dependent enzymeProtein engineeringα,β-Diamino acid

Identifiers

PMID41167839

DOI10.1016/bs.mie.2025.08.007

PMCPMC12614450

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