Two-Phase Fermentation for Isoprenoid Yield
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Solution Overview
Problem
Conventional methods for producing isoprenoids, such as extraction from plants and microbes, and chemical synthesis, face challenges like low yield, high costs, and end-product inhibition, while two-phase fermentation systems with water-immiscible solvents are complex and prone to contamination, making industrial-scale production inefficient.
Innovation Solution
A two-phase fermentation process is developed by adding a water-immiscible organic solvent to an aqueous medium, optimizing the solvent-to-medium ratio, and using an oxygen probe to maintain optimal oxygen tension, facilitating selective extraction and reducing cytotoxic effects, thereby enhancing productivity and yield.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional extraction methods are used to produce isoprenoids, then the production can be achieved, but the yield is low and the production methods are complicated
Solution Approach 1:
The patent changes the fundamental production parameter from extraction to fermentation, using metabolic engineering to transform microorganisms into production factories. This shifts the process from extracting existing compounds to synthesizing new compounds through controlled biological metabolism, dramatically improving yield while simplifying the production workflow
Solution Approach 2:
The patent replaces mechanical extraction processes with biological fermentation processes. Instead of using solvents and mechanical separation to extract isoprenoids from plant material, the system uses genetically modified microorganisms to biosynthesize the compounds, eliminating complex extraction and purification steps
2Productivity
If chemical synthesis methods are used to produce isoprenoids, then the production can be achieved, but the cost is high due to expensive starting materials and extensive purification steps
Solution Approach 1:
The patent implements self-service by enabling microorganisms to autonomously synthesize isoprenoids through their metabolic pathways. The engineered cells use readily available carbon sources from the culture medium to produce the target compounds, eliminating the need for expensive chemical starting materials and multiple purification steps required in traditional chemical synthesis
Solution Approach 2:
The patent fundamentally changes the synthesis approach from chemical to biological, using metabolic engineering to rewire microbial metabolism. This allows the use of cheap carbon sources like glucose instead of expensive chemical precursors, and the biological system naturally handles purification through cellular compartmentalization and secretion
3Productivity
If fermentation is used to produce isoprenoids, then the productivity is improved, but end-product inhibition occurs because the product is cytotoxic to the microorganisms
Solution Approach 1:
The patent applies extraction by using a water-immiscible organic solvent phase to continuously remove the isoprenoid product from the aqueous fermentation medium. This extractive fermentation approach maintains a concentration gradient that drives product partitioning into the organic phase, preventing cytotoxic accumulation in the microbial cells while sustaining high production rates
Solution Approach 2:
The patent introduces a water-immiscible organic solvent as an intermediary phase that mediates between the aqueous fermentation medium and the hydrophobic isoprenoid product. This intermediary facilitates product removal and protection of microorganisms by providing a separate phase for product accumulation, eliminating direct contact between high concentrations of cytotoxic product and living cells
4Productivity
If isoprenoids are produced through fermentation, then the productivity is high, but product loss occurs through evaporation due to the volatile nature of isoprenoids
Solution Approach 1:
The patent uses a water-immiscible organic solvent as an intermediary phase that captures volatile isoprenoid products immediately upon formation. This intermediary phase acts as a trap for volatile compounds, preventing their evaporation into the gas phase while allowing continuous production in the aqueous phase, thereby eliminating product loss through evaporation
Solution Approach 2:
The patent exploits phase transitions by utilizing the partitioning of isoprenoids from the aqueous phase into the organic solvent phase. This phase transfer occurs continuously during fermentation, and the organic phase serves as a condensed reservoir for the volatile product, preventing vaporization and maintaining high productivity without loss
5Productivity
If a water-immiscible organic solvent is added to the fermentation medium, then product extraction is improved, but the system becomes complex and prone to contamination
Solution Approach 1:
The patent merges the fermentation and product extraction processes into a single integrated operation. By adding the water-immiscible organic solvent directly to the fermentation medium, the system combines microbial growth, product synthesis, and product removal in one continuous process, eliminating the need for separate extraction equipment and reducing overall system complexity
Solution Approach 2:
The water-immiscible organic solvent serves multiple functions simultaneously: it acts as an extractant for product removal, a protective barrier against cytotoxicity, and a trap for volatile compounds. This multi-functionality reduces the need for additional process steps and equipment, simplifying the overall fermentation system while improving product recovery
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 method allows for an industrially scalable and robust production of isoprenoids with reduced toxicity and volatility issues, maintaining high productivity and yield by optimizing solvent selection and oxygen management, thus overcoming the limitations of conventional methods.
Implementation Method 1
The target product partitions between the extractant and the aqueous fermentation medium when the two are brought into contact, thereby reducing the concentration of the product in the aqueous medium.
Implementation Method 2
providing the two phase system with an oxygen probe; then performing a calibration of said oxygen probe; then optimizing oxygen tension in said two phase system
Data Source
AI summary
The invention relates to a two phase fermentation process for producing an organic compound, in particular an isoprenoid and to a bioreactor comprising a two phase fermentation system for producing an organic compound.