Mutant Microalgae DGLA Production via Desaturase Block

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Solution Overview

Problem

Current methods for producing dihomo-gamma-linolenic acid (DGLA) face challenges due to low DGLA/AA ratios and susceptibility to oxidation in fungal-derived sources, necessitating a natural, 'green' alternative with improved yield and stability.

Innovation Solution

A mutant strain of microalgae Parietochloris incisa defective in its Δ5 desaturase gene is cultivated, resulting in a significantly higher DGLA/AA ratio and incorporation of β-carotene as a natural antioxidant, reducing oxidation risks.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Quantity of substance

If fungal mutants are used to produce DGLA, then DGLA can be produced, but the DGLA/AA ratio is low (approximately 12) and the product is susceptible to oxidation

Engineering Contradiction:
ImproveDGLA concentrationVSAvoidoxidation stability
Core Design Contradiction:
Quantity of substanceVSReliability

Solution Approach 1:

The patent changes the source organism from fungal mutants to microalgal mutants, fundamentally altering the biochemical pathway and product characteristics. This parameter change enables achieving a DGLA/AA ratio greater than 25 while incorporating natural antioxidants that prevent oxidation

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent converts the potential harm of oxidation susceptibility into a benefit by incorporating β-carotene, a natural antioxidant, into the microalgal cell structure. This natural antioxidant prevents oxidation of the DGLA-rich oil, transforming a vulnerable product into a stable one

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

2Quantity of substance

If wild-type microalgae are cultivated, then AA can be produced, but DGLA accumulation is limited due to functional Δ5 desaturase activity

Engineering Contradiction:
ImproveDGLA accumulationVSAvoidDGLA/AA ratio
Core Design Contradiction:
Quantity of substanceVSProductivity

Solution Approach 1:

The patent extracts or removes the functional Δ5 desaturase enzyme from the microalgal cell through genetic mutation. This extraction prevents the conversion of DGLA to AA, causing DGLA to accumulate in the cell while maintaining high productivity of this valuable fatty acid

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

Instead of using wild-type microalgae that convert DGLA to AA through functional desaturase, the patent inverts the approach by using mutants with defective or absent desaturase activity. This inversion causes the metabolic flux to be blocked at the DGLA stage, resulting in high DGLA accumulation and improved DGLA/AA ratio

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

3Reliability

If synthetic antioxidants are added to fungal-derived PUFAs, then oxidation can be prevented, but the product becomes more complex and less natural

Engineering Contradiction:
Improveoxidation resistanceVSAvoidproduct composition complexity
Core Design Contradiction:
ReliabilityVSDevice complexity

Solution Approach 1:

The patent enables the microalgal cell to serve itself by producing and incorporating β-carotene, a natural antioxidant, within its own structure. This self-service approach provides oxidation protection without requiring external addition of synthetic antioxidants, keeping the product composition simple and natural

Inventive Principle:
Principle #25Self-service

Solution Approach 2:

The patent converts the potential harm of oxidation susceptibility into a benefit by incorporating β-carotene, a natural antioxidant, into the microalgal cell structure. This natural antioxidant prevents oxidation of the DGLA-rich oil, transforming a vulnerable product into a stable one

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 process achieves a DGLA/AA ratio greater than 25, with DGLA concentration exceeding 30% of total fatty acids, and provides a stable, oxidation-resistant DGLA-rich oil suitable for various applications, including animal feeds and health supplements.

Implementation Method 1

a mutant strain of the microalga Parietochloris incisa that is defective in its Δ5 desaturase (Δ5D) gene

Methodology Applied
Scientific EffectEnzyme deficiency (Δ5 desaturase): Enzyme

Implementation Method 2

incorporation of β-carotene as a natural antioxidant, reducing oxidation risks

Methodology Applied
Scientific EffectOxidation resistance: Oxidation

Data Source

PatentUS8871808B2Over-production of dihomo linolenic acid by a mutant strain of Parietochloris incisa
Publication Date: 2014.10.28 BEN GURION UNIVERSITY OF THE NEGEV

AI summary

The present invention is primarily directed to a mutant strain of Parietochloris incisa, characterized in comprising a substantially reduced intracellular concentration of arachidonic acid (AA) together with a substantially increased intracellular concentration of dihomo-gamma-linolenic acid 20:3ω6 (DGLA).