Perfluorocarbon Underlay for Microbial Metabolite Extraction
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Solution Overview
Problem
Current methods for extracting metabolites from microbial cultures using hydrophobic solvents face challenges such as scalability issues, emulsion formation, and safety concerns due to flammability and environmental hazards, particularly when dealing with larger culture volumes and turbid bioreactors.
Innovation Solution
The use of denser-than-water, biocompatible, and non-toxic liquid perfluorocarbons as an underlay in microbial cultures for extracting hydrophobic metabolites, combined with a secondary extraction step using environmentally friendly solvents like ethanol, allows for continuous and non-destructive extraction of metabolites at standard ambient temperature and pressure.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If hydrophobic solvents (decane, dodecane, heptadecane) are used as overlay for metabolite extraction, then extraction efficiency is improved, but emulsion formation occurs limiting lifetime and product extractability
Solution Approach 1:
The patent inverts the traditional overlay approach by using a denser hydrophobic solvent (perfluorocarbon) as an underlay beneath the aqueous culture medium. This inversion prevents emulsion formation because the underlay configuration avoids direct contact between the hydrophobic solvent and turbulent culture broth, while still enabling effective metabolite extraction through the culture medium interface.
Solution Approach 2:
The patent changes the density parameter of the extraction solvent by selecting perfluorocarbons (density 1.5-2.0 g/cm³) that are denser than water, as opposed to traditional lighter hydrocarbons. This parameter change enables the solvent to form a stable underlay layer that remains separated from the culture medium, preventing emulsion formation while maintaining extraction efficiency.
2Productivity
If alkane solvents are used for extraction, then metabolite partitioning is improved, but flammability and compatibility issues with silicone tubing arise
Solution Approach 1:
The patent replaces traditional alkane solvents with perfluorocarbon solvents that have superior safety profiles. Perfluorocarbons are non-flammable and chemically inert, eliminating fire hazards and compatibility issues with silicone-based bioreactor components, while maintaining effective metabolite extraction capabilities.
Solution Approach 2:
The patent utilizes the chemically inert nature of perfluorocarbon solvents to create a safe extraction environment. These solvents do not react with bioreactor materials including silicone tubing, are non-flammable, and provide a stable, inert extraction phase that eliminates safety concerns associated with traditional hydrocarbon solvents.
3Productivity
If hydrophobic solvent overlay is used in turbid bioreactors, then extraction occurs, but turbidity at interface creates emulsion that blows out of reactors or lyses cells
Solution Approach 1:
The patent inverts the extraction interface configuration by placing the hydrophobic solvent underlay beneath the aqueous culture medium instead of as an overlay. This inversion positions the extraction interface at the bottom of the bioreactor where it is not subjected to turbulent mixing, sparging, or foaming, thereby preventing emulsion formation and cell lysis while maintaining extraction efficiency.
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 enables scalable, safe, and efficient extraction of metabolites from microbial cultures, avoiding emulsion issues and maintaining microbial viability, while allowing for high-purity product recovery without the need for pressure gradients or specialized infrastructure.
Implementation Method 1
The use of denser-than-water, biocompatible, and non-toxic liquid perfluorocarbons as an underlay in microbial cultures for extracting hydrophobic metabolites
Implementation Method 2
The use of denser-than-water, biocompatible, and non-toxic liquid perfluorocarbons as an underlay in microbial cultures
Data Source
AI summary
Methods and systems that use fluorocarbons for designing scalable living, in-line extraction of heterologous metabolites from microbial cultures are disclosed. The disclosed systems and methods are organism agnostic and be applied to bacteria, yeasts, filamentous microbes, and algae. The disclosed methods and systems use pperfluorocarbon solvent to extract terpenes from engineered microorganisms and allows non-lethal milking of hydrocarbon products from filamentous and single-celled/multicelled microbial cultures. The disclosed process preferably does not use pressure gradients and relies on the physical interaction of microbes with fluorocarbon at satp and standard temperature of microbial cultivation to partition non-native products. The disclosed process and system also allow for increased production of products providing a physical sink which enables the forward reactions to produce more products than in its absence.