Engineered Transaminase for Chiral Amine Synthesis

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Solution Overview

Problem

Current methods for synthesizing rivastigmine, a drug used to treat Alzheimer's and Parkinson's disease-related dementia, are inefficient, costly, and hazardous, lacking effective transaminase biocatalysts for producing intermediate chiral amine compounds.

Innovation Solution

Engineered transaminase polypeptides with specific amino acid sequence modifications are used to convert 3-hydroxyacetophenone to (S)-3-(1-aminoethyl)-phenol with enhanced activity, enantiomeric excess, and stability, allowing for efficient production of rivastigmine intermediates.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If conventional chemical synthesis methods are used for rivastigmine production, then the process can proceed with existing technology, but the synthesis is inefficient, costly, and hazardous

Engineering Contradiction:

Improvesynthesis efficiencyVSAvoidhazardous conditions

Core Design Contradiction:

ProductivityVSObject-affected harmful factors

Solution Approach 1:

The patent replaces conventional chemical synthesis methods with biocatalytic transaminase enzymes. The transaminase catalysts perform the chemical transformation that previously required hazardous chemical reagents and conditions, thereby eliminating harmful factors while improving synthesis efficiency and productivity

Inventive Principle:

Principle #28Mechanics substitution (Replace mechanical system)

Solution Approach 2:

The patent modifies the reaction parameters by using enzymatic catalysis instead of chemical catalysis. The transaminase enzymes operate under milder conditions with higher selectivity and efficiency, changing the fundamental parameters of the synthesis process to achieve improved productivity and reduced hazards

Inventive Principle:

Principle #35Parameter changes

2Productivity

If transaminase biocatalysts are developed for chiral amine production, then synthesis efficiency and enantiomeric excess improve, but the biocatalyst development and optimization require significant research investment

Engineering Contradiction:

Improvesynthesis efficiencyVSAvoidbiocatalyst complexity

Core Design Contradiction:

ProductivityVSDevice complexity

Solution Approach 1:

The patent segments the biocatalyst development into distinct components: identifying specific amino acid positions (X18, X163, X235, X244, X323, X383, X424, X427) that control enzyme activity and selectivity, and systematically optimizing each position independently to achieve the desired performance without overwhelming complexity

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The patent systematically varies amino acid residues at specific positions in the transaminase sequence to optimize catalytic activity and enantiomeric excess. This parameter-based approach allows controlled improvement of synthesis efficiency while managing biocatalyst complexity through targeted modifications

Inventive Principle:

Principle #35Parameter changes

3Manufacturing precision

If engineered transaminase polypeptides with multiple amino acid modifications are used, then enzymatic activity and enantiomeric excess increase, but the protein sequence complexity and optimization difficulty increase

Engineering Contradiction:

Improveenantiomeric excessVSAvoidpolypeptide sequence complexity

Core Design Contradiction:

Manufacturing precisionVSDevice complexity

Solution Approach 1:

The patent applies local quality by making specific amino acid modifications at predetermined positions (X18, X163, X235, X244, X323, X383, X424, X427) rather than random modifications. Each position is selected based on its specific role in determining enantiomeric excess and catalytic activity, allowing precise control over product quality while managing sequence complexity

Inventive Principle:

Principle #3Local quality

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 engineered transaminase polypeptides demonstrate at least 2-fold increased activity and high enantiomeric excess in converting 3-hydroxyacetophenone to (S)-3-(1-aminoethyl)-phenol, improving the efficiency and cost-effectiveness of rivastigmine production.

Implementation Method 1

engineered transaminase polypeptides with specific amino acid sequence modifications are used to convert 3-hydroxyacetophenone to (S)-3-(1-aminoethyl)-phenol

Methodology Applied

Scientific EffectTransaminase catalysis: Catalysis

Implementation Method 2

The engineered transaminase polypeptides demonstrate at least 2-fold increased activity and high enantiomeric excess in converting 3-hydroxyacetophenone to (S)-3-(1-aminoethyl)-phenol

Methodology Applied

Scientific EffectEnzyme catalysis: Enzyme

Data Source

PatentEP2582799B1Biocatalysts and methods for the synthesis of (s)-3-(1-aminoethyl)-phenol

Publication Date: 2017.12.20 CODEXIS INC

AI summary

The present disclosure provides engineered transaminase polypeptides having improved properties as compared to naturally occurring transaminases including the ability of converting the substrate, 3'-hydroxyacetophenone to (S)-3-(1-aminoethyl)-phenol in enantiomeric excess and high percentage conversion. Also provided are polynucleotides encoding the engineered transaminases, host cells capable of expressing the engineered transaminases, and methods of using the engineered transaminases to synthesize (S)-3-(1-aminoethyl)-phenol and related compounds useful in the production of active pharmaceutical ingredients.