DAHP Synthase Variant for oAB Fermentation Yield

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Solution Overview

Problem

Existing methods for producing ortho-aminobenzoic acid (oAB) through microbial fermentation are not efficiently improved by using known variants of 3-deoxyarabinoheptulosanate 7-phosphate synthase, particularly in Corynebacterium glutamicum.

Innovation Solution

Utilization of a specific variant of 3-deoxyarabinoheptulosanate 7-phosphate synthase with an amino acid sequence defined by SEQ ID NO. 11 or its variants, which is recombinantly expressed in microbial cells and cultured under conditions that enhance metabolic activity, to improve the production of oAB in microbial fermentation.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If known variants of 3-deoxyarabinoheptulosanate 7-phosphate synthase are used for oAB production in Corynebacterium glutamicum, then the production efficiency is limited, but using the specific variant from EP 0 077 196 shows surprising improvement

Engineering Contradiction:

ImproveoAB production efficiencyVSAvoidenzyme performance consistency

Core Design Contradiction:

ProductivityVSReliability

Solution Approach 1:

The patent applies parameter changes by modifying the amino acid sequence of DAHP synthase based on the variant from EP 0 077 196, specifically adapting it for Corynebacterium glutamicum expression. The sequence modifications (SEQ ID NO: 11 and variants) change the enzyme's catalytic parameters to improve oAB production efficiency while maintaining functional reliability in the new host system.

Inventive Principle:

Principle #35Parameter changes

2Productivity

If DAHP synthase activity is increased to improve oAB production, then metabolite flow through shikimate pathway increases, but enzyme inhibition by aromatic amino acids limits further improvement

Engineering Contradiction:

ImproveoAB production rateVSAvoidaromatic amino acid inhibition

Core Design Contradiction:

ProductivityVSObject-affected harmful factors

Solution Approach 1:

The patent converts the harmful inhibition effect of aromatic amino acids into a benefit by selecting and optimizing a DAHP synthase variant that exhibits reduced sensitivity to this inhibition. The enzyme variant from EP 0 077 196, when expressed in C. glutamicum, maintains high activity even in the presence of aromatic amino acids, effectively turning the inhibitory environment into a favorable condition for sustained high-level oAB production.

Inventive Principle:

Principle #22Blessing in disguise (Convert harm into benefit)

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 use of this enzyme variant significantly increases the production of oAB, achieving extracellular concentrations of at least 1 g/L, preferably 2 g/L, and most preferably 4 g/L, thereby enhancing the efficiency and yield of oAB production in microbial fermentation.

Implementation Method 1

3-Deoxyarabinoheptulosanate 7-phosphate synthase (DAHP synthase) is an enzyme that catalyzes the condensation of phosphoenolpyruvate and erythrose 4-phosphate to form 3-deoxyarabinoheptulosanate 7-phosphate

Methodology Applied

Scientific EffectEnzyme catalysis: Enzyme

Implementation Method 2

producing ortho-aminobenzoic acid (oAB) by microbial fermentation

Methodology Applied

Scientific EffectFermentation: Fermentation

Data Source

PatentUS20250043254A1Particularly suitable 3-desoxyarabinoheptulosanate-7-phosphate synthase for the fermentative preparation of ortho-aminobenzoic acid

Publication Date: 2025.02.06 COVESTRO DEUTSCHLAND AG

AI summary

The present invention relates to the use of specific variants of 3-deoxyarabinoheptulosanate 7-phosphate synthase for producing ortho-aminobenzoic acid by means of microbial fermentation and microorganisms suitable for this purpose.