Mucor circinelloides Recombinant Strain for SDA Production
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Solution Overview
Problem
Current methods for producing stearidonic acid (SDA) are limited due to high fermentation costs and low biomass yield in Mortierella alpina, while Mucor circinelloides has high biomass but lacks SDA production capabilities, necessitating a strategy to enhance SDA production efficiently.
Innovation Solution
The introduction of the Δ15 desaturase gene from Mortierella alpina into Mucor circinelloides via homologous recombination to create a recombinant strain capable of producing SDA, with optimized fermentation conditions using safflower or sunflower seed oil, resulting in a SDA content of up to 8.3% of total fatty acids.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If Mortierella alpina is used for SDA production, then SDA content can be produced, but fermentation cost is high and biomass yield is low
Solution Approach 1:
The patent combines the beneficial traits of two different microorganisms: Mucor circinelloides provides high biomass yield and easy cultivation, while Mortierella alpina provides the Δ15 desaturase enzyme capability for SDA production. By transferring the Δ15 desaturase gene from M. alpina into M. circinelloides through homologous recombination, the invention merges the high biomass advantage of M. circinelloides with the SDA synthesis capability of M. alpina, resolving the contradiction between SDA production and biomass yield.
Solution Approach 2:
The patent uses an intermediary approach by introducing the Δ15 desaturase gene from M. alpina as a foreign gene into the M. circinelloides host. This gene acts as an intermediary element that enables the host organism to perform the SDA synthesis function it naturally lacks, while maintaining the host's advantageous characteristics of high biomass and ease of cultivation.
2Quantity of substance
If Mortierella alpina is used for SDA production, then SDA content can be produced, but fermentation cost is high
Solution Approach 1:
The patent merges the cost-effective cultivation characteristics of Mucor circinelloides with the SDA production capability of Mortierella alpina. M. circinelloides is known for its ease of cultivation and lower fermentation costs compared to M. alpina, while the transferred Δ15 desaturase gene provides the necessary SDA synthesis function, thereby reducing fermentation costs while maintaining SDA production.
3Productivity
If Mucor circinelloides is used for SDA production, then biomass is high, but SDA production capability is lacking
Solution Approach 1:
The patent introduces the Δ15 desaturase gene from M. alpina as an intermediary genetic element into M. circinelloides. This foreign gene serves as the missing functional component that enables SDA production in the host, while the host's high biomass production capability remains intact. The gene transfer approach allows M. circinelloides to simultaneously achieve high biomass and SDA production.
Solution Approach 2:
The patent transforms Mucor circinelloides from a single-function organism (high biomass production only) into a multi-functional organism that can both produce high biomass and synthesize SDA. By introducing the Δ15 desaturase gene, the host organism gains the additional capability of SDA production while maintaining its original high biomass production function, achieving multi-functionality.
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 recombinant Mucor circinelloides strain Mc-Δ15 achieves a significant increase in SDA content, reaching 5.2% to 8.3% of total fatty acids, providing a viable industrial solution for SDA production.
Implementation Method 1
Stearidonic acid SDA (18:4n-3) is synthesized by α-linolenic acid ALA (18:3n-3) under the catalysis of Δ6 desaturase, or γ-linolenic acid (GLA, 18:3, n-6) under the catalysis of Δ15 Des.
Implementation Method 2
Δ15 Des is capable of catalyzing GLA (18:3, n-6) to produce SDA
Implementation Method 3
fermentation technology thereof, including that Δ15 Des obtained by cloning from a Mortierella alpina strain is transferred into Mucor circinelloides by a homologous recombination method
Data Source
AI summary
Provided is construction of Mucor circinelloides cell factory for producing stearidonic acid (SDA) and fermentation technology. Δ15 Desaturase gene is obtained by cloning from Mortierella alpina, the gene is ligated to an integrative plasmid pMAT1552, and transformed into a Mucor circinelloides defective strain Mu402, and Δ15 Desaturase gene is integrated on Mucor circinelloides genome through homologous recombination, to obtain the recombinant strain Mc-Δ15, finally, the expression of the Δ15 Desaturase gene in Mucor circinelloides is realized. The recombinant new strain is accession number CGMCC No. 15888, and the classification name is Mucor circinelloides-D15D.