Mutated Esterase for Non-Natural Substrate Selectivity

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Solution Overview

Problem

Wild-type biocatalysts exhibit unsatisfactory reactivity, stability, and selectivity for non-natural substrates, limiting their effectiveness in organic synthesis, particularly in producing chiral compounds with high chemical and regional selectivity.

Innovation Solution

A site-directed mutagenesis approach is applied to an esterase enzyme, introducing specific mutations such as N51G and combinations thereof, enhancing its substrate binding and catalytic efficiency, thereby improving enzyme specificity and activity.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Adaptability or versatility

If wild-type esterase is used for catalysis, then the enzyme has good reactivity for natural substrates, but the reactivity, stability and selectivity for non-natural substrates are unsatisfactory

Engineering Contradiction:

Improvesubstrate scopeVSAvoidreactivity and selectivity

Core Design Contradiction:

Adaptability or versatilityVSReliability

Solution Approach 1:

The patent applies site-directed mutagenesis to change specific amino acid residues (N51G, M52F/L/N/Y/G/W, W115P, T117L/M/F/W/A/I, S140A/G/N/C/T/V/L/P, A142V/L/P/S, V167M, I195L/F/T/V, W196I/L/V, D217M/Q/A/S/G, L231T/I, V267E/C/I/V, S295T/A/Y/F/M/N) in the esterase protein structure. These parameter changes at the molecular level modify the enzyme's active site characteristics, enabling it to accommodate and catalyze non-natural substrates while maintaining or improving reactivity and selectivity

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The patent introduces different amino acid mutations at specific local positions (residues 51-52, 115, 117, 140, 142, 167, 195-196, 217, 231, 267, 295) of the esterase molecule. Each mutation targets a specific local region to optimize substrate binding and catalytic function, creating localized improvements in enzyme performance for non-natural substrates

Inventive Principle:

Principle #3Local quality

2Productivity

If more enzyme is used to improve reaction efficiency, then the production cost increases

Engineering Contradiction:

Improvereaction efficiencyVSAvoidenzyme usage

Core Design Contradiction:

ProductivityVSQuantity of substance

Solution Approach 1:

The patent modifies the enzyme's catalytic parameters through amino acid substitution, creating variants with enhanced specific activity. The mutated esterase demonstrates significantly improved catalytic efficiency toward non-natural substrates, allowing reduced enzyme dosage while maintaining or improving productivity

Inventive Principle:

Principle #35Parameter changes

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

The mutated esterase demonstrates significantly improved specificity and activity, reducing enzyme usage and production costs while achieving high enantiomeric excess in the production of chiral acids.

Implementation Method 1

Esterase refers to the general name of a class of enzymes with the ability to hydrolyze an ester bond

Methodology Applied

Scientific EffectHydrolysis: Hydrolysis

Implementation Method 2

enhancing its substrate binding and catalytic efficiency, thereby improving enzyme specificity and activity

Methodology Applied

Scientific EffectEnzyme-substrate binding: Enzyme

Data Source

PatentUS12516303B2Esterase mutant and use thereof

Publication Date: 2026.01.06 ASYMCHEM LIFE SCI TIANJIN

AI summary

Provided are an esterase mutant and use thereof. The amino acid sequence of the esterase mutant has a sequence as shown in SEQ ID NO: 1, and sites at which amino acid mutations occur include an N51G site.