Recombinant Bacteria Co-Expressing G6PD and MDH for Organic Acid Production

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Solution Overview

Problem

Current methods for producing organic acids, such as succinic acid, face challenges in achieving high titer, productivity, and yield, which are crucial for economic viability in industrial applications, and existing technologies do not effectively optimize these parameters simultaneously.

Innovation Solution

Recombinant bacteria expressing heterologous glucose-6-phosphate dehydrogenase and malate dehydrogenase enzymes, either natively or through overexpression, are used to enhance organic acid production, specifically by co-expressing these enzymes in a single fermentation process to promote higher yields and productivity.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If conventional fermentation methods are used for organic acid production, then the process is simple to operate, but the titer, productivity, and yield are insufficient for economic viability

Engineering Contradiction:
Improveorganic acid production efficiencyVSAvoidfermentation process complexity
Core Design Contradiction:
ProductivityVSDevice complexity

Solution Approach 1:

The patent applies parameter changes by modifying the genetic composition of the microorganism through recombinant DNA technology. Specifically, it introduces heterologous genes encoding glucose-6-phosphate dehydrogenase and malate dehydrogenase enzymes, and optimizes the expression levels of native enzymes involved in the succinate fermentation pathway. This genetic parameter modification enables the microorganism to achieve high titer, high productivity, and high yield of organic acid production while maintaining a relatively simple fermentation process.

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent uses recombinant microorganisms as intermediaries to bridge the gap between simple fermentation processes and high productivity requirements. The genetically modified microorganism acts as a biological catalyst that converts substrate to organic acid with enhanced efficiency, thereby achieving economic viability without complicating the overall fermentation process structure.

Inventive Principle:
Principle #24Intermediary (Mediator)

2Productivity

If high titer and high productivity are achieved through conventional means, then production efficiency improves, but yield and economic viability remain compromised

Engineering Contradiction:
Improveorganic acid production rateVSAvoidorganic acid yield
Core Design Contradiction:
ProductivityVSQuantity of substance

Solution Approach 1:

The patent simultaneously optimizes multiple metabolic parameters by introducing heterologous enzymes and overexpressing key pathway enzymes. The glucose-6-phosphate dehydrogenase increases NADPH availability, while malate dehydrogenase enhances the reductive branch of the TCA cycle. This coordinated parameter optimization ensures that high productivity is achieved without compromising yield, as both titer and productivity are enhanced through the same genetic modifications.

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 approach results in improved organic acid production, achieving high concentrations and productivity rates, thereby optimizing the economic viability of the fermentation process by balancing titer, productivity, and yield.

Implementation Method 1

expressing heterologous glucose-6-phosphate dehydrogenase or overexpressing glucose-6-phosphate dehydrogenase

Methodology Applied
Scientific EffectEnzyme catalysis: Enzyme

Implementation Method 2

expressing heterologous malate dehydrogenase or overexpressing malate dehydrogenase

Methodology Applied
Scientific EffectEnzyme catalysis: Enzyme

Implementation Method 3

producing organic acids... culturing the recombinant bacteria with a carbon source under conditions favoring organic acid production

Methodology Applied
Scientific EffectFermentation: Fermentation

Data Source

PatentEP2904104B1Recombinant microorganisms for producing organic acids
Publication Date: 2018.08.15 MICHIGAN BIOTECHNOLOGY INSTITUTE
  • EP2904104B1 patent drawingFigure 1
  • EP2904104B1 patent drawingFigure 2
  • EP2904104B1 patent drawingFigure 3

AI summary

Recombinant microorganisms that co express enzymatic glucose-6-phosphate dehydrogenase and malate dehydrogenase are generated to produce organic acids.