Corynebacterium glutamicum Transformant for 4-ABA Production

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Solution Overview

Problem

Current methods for producing 4-aminobenzoic acid (4-ABA) are inefficient and environmentally unsustainable, relying on fossil resources and requiring high energy, while existing biorefinery techniques using microorganisms yield low productivity and are not practical for industrial-scale production.

Innovation Solution

A transformant of Corynebacterium glutamicum is engineered to produce 4-ABA by introducing genes encoding para-aminobenzoate synthase component I, para-aminobenzoate synthase component II, and 4-amino-4-deoxychorismate lyase, allowing for high-yield production from saccharides under aerobic conditions, with the bacterium exhibiting high resistance to 4-ABA, thus enabling efficient and sustainable production.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Ease of manufacture

If conventional multi-phase reactions using petroleum are used to synthesize 4-ABA, then the production pathway is established, but massive energy is needed and dependence on fossil resources continues

Engineering Contradiction:
Improveproduction pathwayVSAvoidenergy consumption
Core Design Contradiction:
Ease of manufactureVSUse of energy by moving object

Solution Approach 1:

The patent replaces the conventional chemical synthesis pathway (mechanical/chemical system requiring strong acids and high temperature) with a biological system using genetically modified microorganisms that catalyze 4-ABA production through enzymatic reactions under mild conditions, thereby substituting energy-intensive chemical processes with biologically-driven reactions

Inventive Principle:
Principle #28Mechanics substitution (Replace mechanical system)

Solution Approach 2:

The patent changes the fundamental parameters of the production system by shifting from petroleum-based chemical synthesis (high temperature, strong acids, multiple phases) to sugar-based biological fermentation (ambient temperature, physiological pH, single phase), fundamentally altering the energy profile and sustainability of the process

Inventive Principle:
Principle #35Parameter changes

2Object-affected harmful factors

If biorefinery techniques using microorganisms are used to produce 4-ABA, then environmental sustainability is improved, but productivity is low and not practical for industrial production

Engineering Contradiction:
Improveenvironmental impactVSAvoid4-ABA productivity
Core Design Contradiction:
Object-affected harmful factorsVSProductivity

Solution Approach 1:

The patent segments the 4-ABA biosynthesis pathway into distinct enzymatic steps (chorismate → 4-amino-4-deoxychorismate via PabA/PabB, then 4-amino-4-deoxychorismate → 4-ABA via PabC) and introduces specific genes encoding these enzymes into host microorganisms, enabling controlled and optimized production of each step to achieve high overall productivity

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The patent creates composite biological systems by combining multiple genetic elements (pabA, pabB, and pabC genes from different sources) into a single engineered microorganism, creating a composite enzymatic system that works synergistically to achieve high 4-ABA productivity while maintaining environmental sustainability

Inventive Principle:
Principle #40Composite materials

3Reliability

If strong acids and high temperature conditions are used in synthesis reactions, then the chemical pathway proceeds, but massive energy is needed

Engineering Contradiction:
Improvereaction pathwayVSAvoidenergy requirement
Core Design Contradiction:
ReliabilityVSUse of energy by stationary object

Solution Approach 1:

The patent replaces harsh chemical reaction conditions (strong acids, high temperature) with biological catalysis using engineered microorganisms that perform the same chemical transformations under physiological conditions, eliminating the need for energy-intensive equipment and processes while maintaining reaction reliability

Inventive Principle:
Principle #28Mechanics substitution (Replace mechanical system)

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 Corynebacterium glutamicum transformant achieves high concentration and yield of 4-ABA, reducing production costs and environmental impact, making it suitable for industrial applications as a raw material for polymers and pharmaceuticals.

Implementation Method 1

a para-aminobenzoate synthase component I (PabB) synthesizes 4-amino-4-deoxychorismate from the ammonium group and chorismate

Methodology Applied
Scientific EffectEnzymatic reaction: Enzyme

Implementation Method 2

4-amino-4-deoxychorismate is converted to 4-ABA by a 4-amino-4-deoxychorismate lyase (PabC)

Methodology Applied
Scientific EffectEnzymatic reaction: Enzyme

Implementation Method 3

The 4-ABA productivity thereof, however, is 35 mM (4.8 g/L) per 48 hours, which is low, and is unsatisfactory for the industrial production. Still further, Patent Document 1 reports the production of 4-ABA by using a transformant obtained by introducing pabAB derived from Saccharopolyspora erythraea into Streptomyces lividans

Methodology Applied
Scientific EffectAerobic metabolism: Aerobic Digestion

Data Source

PatentEP3421599B1Coryneform bacterial transformant and method for producing 4-aminobenzoic acid or salt thereof using same
Publication Date: 2021.05.05 RES INST OF INNOVATIVE TECH FOR THE EARTH
  • EP3421599B1 patent drawingFigure 1
  • EP3421599B1 patent drawingFigure 2~3
  • EP3421599B1 patent drawing

AI summary

Provided is a microorganism that is capable of efficiently producing para-aminobenzoic acid (4-ABA) or a salt thereof, using saccharides as raw materials, and a method for efficiently producing 4-ABA or a salt thereof by using this microorganism. A transformant obtained by introducing, into a coryneform bacterium, a gene that encodes 4-amino-4-deoxychorismate lyase, a gene that encodes a para-aminobenzoate synthase component I, and a gene that encodes a para-aminobenzoate synthase component II, is capable of efficiently producing 4-ABA or a salt thereof from saccharides.