Engineered Microorganisms for Fatty Acid Chain Length Control

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Solution Overview

Problem

Current methods for producing fatty acids and fatty acid derivatives face challenges due to the need for costly feedstock processing, volatile feedstock prices, and sustainability concerns, particularly with petrochemical and oleochemical sources, which require more specific and sustainable production methods.

Innovation Solution

Genetically modified organisms are developed to produce specific fatty acid chain lengths by encoding heterologous nucleic acid sequences for enzymes like 3-ketoacyl-CoA synthase, ketoacyl-CoA reductase, hydroxyacyl-CoA dehydratase, and enoyl-CoA reductase, enabling the production of fatty acids and derivatives with controlled carbon chain lengths using malonyl-CoA as a substrate.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Quantity of substance

If petrochemical or oleochemical feedstocks are used to produce fatty acids, then production volume is achieved, but feedstock price volatility and sustainability concerns worsen

Engineering Contradiction:
Improvefatty acid production volumeVSAvoidfeedstock price stability
Core Design Contradiction:
Quantity of substanceVSReliability

Solution Approach 1:

The microorganism is engineered to produce fatty acids autonomously using readily available carbon sources like glucose. The metabolic pathway is internally configured to convert carbon sources into specific chain length fatty acids through controlled elongation cycles, eliminating dependence on volatile petrochemical or oleochemical markets

Inventive Principle:
Principle #25Self-service

Solution Approach 2:

The invention changes the production parameter from feedstock-based to microorganism-based synthesis. By manipulating genetic expression of elongation enzymes and controlling cultural conditions, the system produces fatty acids with specific chain lengths directly, transforming the production paradigm away from feedstock processing

Inventive Principle:
Principle #35Parameter changes

2Quantity of substance

If oleochemical feedstocks are used to produce fatty acids, then production volume is achieved, but sustainability performance worsens due to deforestation concerns

Engineering Contradiction:
Improvefatty acid production volumeVSAvoidenvironmental impact from deforestation
Core Design Contradiction:
Quantity of substanceVSObject-affected harmful factors

Solution Approach 1:

The microorganism autonomously synthesizes fatty acids using carbon sources such as glucose, eliminating the need for palm oil or other oleochemical feedstocks associated with deforestation. The system serves itself by converting simple carbon sources into complex fatty acid structures through engineered metabolic pathways

Inventive Principle:
Principle #25Self-service

Solution Approach 2:

The invention replaces sustainable-but-controversial oleochemical feedstocks with renewable carbon sources like glucose that have minimal environmental footprint. The microorganism consumes these short-lived carbon sources to produce fatty acids, avoiding the long-term environmental damage of deforestation

Inventive Principle:
Principle #27Cheap short-living objects (Disposable)

3Manufacturing precision

If conventional feedstock processing methods are used to isolate specific fatty acid chain lengths, then fatty acid purity is improved, but production cost worsens due to fractionation and distillation requirements

Engineering Contradiction:
Improvefatty acid chain length specificityVSAvoidproduction cost
Core Design Contradiction:
Manufacturing precisionVSEase of manufacture

Solution Approach 1:

The microorganism is pre-engineered with specific metabolic pathways that directly produce fatty acids of desired chain lengths during growth. By controlling the expression of elongation enzymes and cultural conditions beforehand, the system synthesizes target fatty acids selectively, eliminating the need for post-production separation processes

Inventive Principle:
Principle #10Preliminary action

Solution Approach 2:

Instead of producing a mixture of fatty acids and then separating them through fractionation and distillation, the invention inverts the approach by engineering the microorganism to produce only the desired chain length fatty acids directly. This reverses the traditional sequence from separation-after-production to selective-production-before-separation

Inventive Principle:
Principle #13The other way round (Inversion)

4Manufacturing precision

If conventional feedstock processing methods are used to isolate specific fatty acid chain lengths, then fatty acid purity is improved, but production complexity worsens due to multiple processing steps

Engineering Contradiction:
Improvefatty acid chain length specificityVSAvoidprocessing procedure complexity
Core Design Contradiction:
Manufacturing precisionVSDevice complexity

Solution Approach 1:

The invention extracts and eliminates the complex fractionation and distillation steps from the production process. By engineering the microorganism to produce specific chain length fatty acids selectively, the system removes the need for downstream separation equipment and multiple processing stages, simplifying the overall manufacturing flow

Inventive Principle:
Principle #2Taking out (Extraction)

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 allows for the efficient and sustainable production of fatty acids and derivatives with high specificity and purity, reducing production costs and environmental impact by utilizing renewable resources and minimizing the need for costly feedstock processing.

Implementation Method 1

a 3-ketoacyl-CoA synthase, capable of catalyzing a condensation reaction to condense malonyl-CoA with an acyl-CoA to form a 3-ketoacyl-CoA

Methodology Applied
Scientific EffectCondensation reaction:

Implementation Method 2

a ketoacyl-CoA reductase, capable of catalyzing a reduction reaction to reduce a 3-keto group in the 3-ketoacyl-CoA to a hydroxyl group to form a 3-hydroxyacyl-CoA

Methodology Applied
Scientific EffectReduction reaction: Reduction

Implementation Method 3

a hydroxyacyl-CoA dehydratase, capable of catalyzing a dehydration reaction to remove water from the 3-hydroxyacyl-CoA to form an enoyl-CoA

Methodology Applied
Scientific EffectDehydration reaction:

Implementation Method 4

an enoyl-CoA reductase capable of catalyzing a reduction reaction to reduce an enoyl group in the enoyl-CoA to form an acyl-CoA

Methodology Applied
Scientific EffectReduction reaction: Reduction

Data Source

PatentEP3022310B1Microorganisms and methods for the production of fatty acids and fatty acid derived products
Publication Date: 2019.10.16 CARGILL INC
  • EP3022310B1 patent drawingFigure 1
  • EP3022310B1 patent drawingFigure 2
  • EP3022310B1 patent drawingFigure 3

AI summary

This invention relates to metabolically engineered microorganism strains, such as bacterial strains, in which there is an increased utilization of malonyl-CoA for production of a fatty acid or fatty acid derived product, wherein the modified microorganism produces fatty acyl-CoA intermediates via a malonyl-CoA dependent but malonyl-ACP independent mechanism.