PheP Symporter E272K Variant for Aromatic Amino Acid Production

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Solution Overview

Problem

Existing methods for producing aromatic amino acids, such as phenylalanine and tryptophan, in microorganisms like Escherichia coli and Corynebacterium, do not effectively leverage the potential of phenylalanine:H+ symporter PheP for enhanced productivity.

Innovation Solution

A variant of phenylalanine:H+ symporter PheP is developed by substituting glutamic acid at position 272 with lysine, resulting in a variant with increased activity, which is integrated into recombinant strains to enhance L-aromatic amino acid production.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If wild-type strains or conventional mutant strains are used for aromatic amino acid production, then production capability is maintained, but productivity is insufficient and cannot be significantly improved

Engineering Contradiction:

Improvearomatic amino acid productivityVSAvoidproduction stability

Core Design Contradiction:

ProductivityVSReliability

Solution Approach 1:

The invention applies parameter changes by substituting specific amino acids in the PheP transporter protein sequence (e.g., E272K substitution) to modify its functional properties. This changes the transporter's activity parameters, enabling enhanced phenylalanine uptake capability while maintaining protein stability and function, thereby resolving the contradiction between improving productivity and maintaining reliability

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The invention applies local quality by making targeted amino acid substitutions at specific positions (e.g., position 272) within the PheP protein sequence rather than global modifications. This localized change optimizes the transporter's function in specific regions while preserving the overall protein structure and stability, achieving enhanced productivity without compromising production reliability

Inventive Principle:

Principle #3Local quality

2Productivity

If multiple proteins (enzymes, transcription factors, transport proteins) are targeted to increase L-aromatic amino acid productivity, then productivity may improve, but research complexity and time required increase significantly

Engineering Contradiction:

ImproveL-aromatic amino acid productivityVSAvoidresearch complexity

Core Design Contradiction:

ProductivityVSDevice complexity

Solution Approach 1:

The invention applies the taking out principle by extracting and focusing on a single key limiting factor - the phenylalanine transporter PheP - rather than simultaneously modifying multiple proteins. By identifying and optimizing this specific transporter protein through amino acid substitution, the invention simplifies the research approach while achieving significant productivity improvements in L-aromatic amino acid production

Inventive Principle:

Principle #2Taking out (Extraction)

Data Source

PatentEP4610350A1Novel phenylalanine:h+ symporter phep variant, and method for producing l-aromatic amino acids using same

Publication Date: 2025.09.03 DAESANG CORP

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AI summary

The present invention relates to a novel variant of phenylalanine:H+ symporter PheP and a method of producing L-aromatic amino acids using the same. The phenylalanine:H+ symporter PheP variant is obtained by substituting one or more amino acids in the amino acid sequence constituting phenylalanine:H+ symporter PheP to change the activity of the protein, and a recombinant microorganism comprising the variant is capable of efficiently producing L-tryptophan, L-phenylalanine, or L-tyrosine.