Microbial Organisms with Reductive TCA Pathway Enzymes for Acetyl-CoA Flux

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Solution Overview

Problem

Current methods for producing chemicals like 1,3-butanediol, 4-hydroxybutanoic acid, and isopropanol rely on petroleum-based feedstocks, which are costly and unsustainable, and do not efficiently utilize renewable carbon sources.

Innovation Solution

Engineering non-naturally occurring microbial organisms with enhanced carbon fixation pathways, such as the reductive TCA cycle and Wood-Ljungdahl pathway, to increase carbon flux through acetyl-CoA, allowing for the production of these chemicals from carbohydrate-based carbon feedstocks and syngas components like CO and H2.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If petroleum-based feedstocks are used for chemical production, then current production methods can be maintained, but sustainability and cost are worsened

Engineering Contradiction:

Improvechemical production efficiencyVSAvoidsustainability

Core Design Contradiction:

ProductivityVSReliability

Solution Approach 1:

The patent changes the fundamental parameter of carbon source from petroleum-based to renewable carbohydrate-based feedstocks. By introducing exogenous nucleic acids encoding reductive TCA pathway enzymes into microbial organisms, the metabolic pathways are altered to enable efficient conversion of renewable carbon sources into target chemicals, thereby improving sustainability without compromising production efficiency

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The patent replaces petrochemical conversion processes with biologically-based metabolic pathways. Microbial organisms engineered with exogenous genes perform the chemical synthesis function that previously required petroleum refining and petrochemical processing, substituting a biological system for a mechanical/chemical industrial process

Inventive Principle:

Principle #28Mechanics substitution (Replace mechanical system)

2Reliability

If conventional metabolic pathways are used in microorganisms, then existing production systems can be maintained, but carbon flux through acetyl-CoA is insufficient for efficient chemical production

Engineering Contradiction:

Improveproduction system stabilityVSAvoidcarbon flux efficiency

Core Design Contradiction:

ReliabilityVSProductivity

Solution Approach 1:

The patent segments the metabolic pathway by introducing exogenous nucleic acids that encode specific reductive TCA pathway enzymes. This adds distinct enzymatic steps to the existing microbial metabolism, creating a segmented pathway that efficiently channels carbon flux through acetyl-CoA to produce target chemicals while maintaining overall system stability

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The patent dynamically enhances microbial metabolism by introducing exogenous genetic material that can be regulated and optimized. The engineered pathways allow for dynamic control of carbon flux distribution, enabling the system to adaptively route metabolites toward desired products while maintaining cellular homeostasis

Inventive Principle:

Principle #15Dynamics

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

This approach significantly enhances the yield and efficiency of producing chemicals like 1,3-butanediol, 4-hydroxybutanoic acid, and isopropanol by utilizing renewable carbon sources, reducing reliance on petroleum-based feedstocks and improving product yields from carbohydrate-based carbon feedstocks.

Implementation Method 1

enhance carbon fixation capabilities

Methodology Applied

Scientific EffectCarbon fixation: Photosynthesis

Implementation Method 2

reductive TCA pathway which includes at least one exogenous nucleic acid encoding a reductive TCA pathway enzyme

Methodology Applied

Scientific EffectReductive TCA cycle:

Implementation Method 3

production of these chemicals from carbohydrate-based carbon feedstocks and syngas components like CO and H2

Methodology Applied

Scientific EffectSyngas conversion:

Data Source

PatentUS8048661B2Microbial organisms comprising exogenous nucleic acids encoding reductive TCA pathway enzymes

Publication Date: 2011.11.01 GENOMATICA INC

AI summary

A non-naturally occurring microbial organism includes a microbial organism having a reductive TCA or Wood-Ljungdahl pathway in which at least one exogenous nucleic acid encoding these pathway enzymes is expressed in a sufficient amount to enhance carbon flux through acetyl-CoA. A method for enhancing carbon flux through acetyl-CoA includes culturing theses non-naturally occurring microbial organisms under conditions and for a sufficient period of time to produce a product having acetyl-CoA as a building block. Another non-naturally occurring microbial organism includes at least one exogenous nucleic acid encoding an enzyme expressed in a sufficient amount to enhance the availability of reducing equivalents in the presence of carbon monoxide or hydrogen, thereby increasing the yield of redox-limited products via carbohydrate-based carbon feedstock. A method for enhancing the availability of reducing equivalents in the presence of carbon monoxide or hydrogen includes culturing this organism for a sufficient period of time to produce a product.