Two-Stage Fermentation for Higher Alcohol Production

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Solution Overview

Problem

Current biological methods for producing higher alcohols like isobutanol and n-butanol face challenges due to low yields and toxicity issues, which limit their scalability and efficiency in fermentation processes.

Innovation Solution

A two-stage reaction sequence using different biological catalyst systems in separate bioreactors, where the first stage produces a nontoxic intermediate product that is then converted into the final alcohol in the second stage, allowing for high concentrations and improved reaction kinetics.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If a single-stage fermentation process is used to produce higher alcohols, then the process is simple and easy to operate, but the yield is low and product toxicity limits final concentration

Engineering Contradiction:
Improveyield of higher alcoholsVSAvoidcomplexity of fermentation process
Core Design Contradiction:
ProductivityVSDevice complexity

Solution Approach 1:

The fermentation process is divided into two distinct stages: a first bioreactor produces a nontoxic intermediate product (such as amino acids or organic acids), and a second bioreactor converts this intermediate into the desired higher alcohol. This segmentation allows each stage to be optimized independently, avoiding the toxicity limitations that constrain single-stage processes while achieving higher overall yields.

Inventive Principle:
Principle #1Segmentation

2Quantity of substance

If high concentrations of higher alcohols are produced in a single bioreactor, then productivity increases, but cell growth is inhibited and metabolic pathways are blocked due to toxicity

Engineering Contradiction:
Improveconcentration of higher alcoholVSAvoidcell viability and metabolic function
Core Design Contradiction:
Quantity of substanceVSReliability

Solution Approach 1:

A nontoxic intermediate product serves as a mediator between the first bioreactor and the second bioreactor. The intermediate (such as amino acids or organic acids) can be accumulated to high concentrations without toxic effects on the cells in the first bioreactor. In the second bioreactor, this intermediate is then converted to the higher alcohol, allowing high final concentrations to be achieved while maintaining cell viability throughout the process.

Inventive Principle:
Principle #24Intermediary (Mediator)

3Productivity

If enzymes with high specific activity are engineered into the organism, then production rate increases, but inherent metabolic bottlenecks still limit the concentration of butanol precursors

Engineering Contradiction:
Improverate of alcohol productionVSAvoidconcentration of butanol precursors
Core Design Contradiction:
ProductivityVSQuantity of substance

Solution Approach 1:

The metabolic pathway is segmented across two bioreactors. The first bioreactor is optimized to produce high concentrations of intermediate precursors using engineered enzymes with high specific activity. The second bioreactor then converts these precursors to the final alcohol product. This segmentation removes the metabolic bottleneck by allowing precursor accumulation in the first stage without being limited by the capacity of subsequent enzymatic steps.

Inventive Principle:
Principle #1Segmentation

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 efficient and scalable production of branched-chain alcohols like isobutanol, n-butanol, 3-methyl-1-butanol, and 2-methyl-1-butanol, overcoming toxicity issues and achieving higher yields by segregating the production stages.

Implementation Method 1

providing a first bioreactor with a first biological catalyst system and converting in the first bioreactor a feedstock into an intermediate product comprising an amino acid or lactic acid

Methodology Applied
Scientific EffectFermentation: Fermentation

Implementation Method 2

providing a second bioreactor with a second biological catalyst system and converting in the second bioreactor at least a portion of the intermediate product into said alcohol

Methodology Applied
Scientific EffectFermentation: Fermentation

Data Source

PatentEP2885415B1Two stage production of higher alcohols
Publication Date: 2022.08.03 EASEL BIOTECHNOLOGIES LLC
  • EP2885415B1 patent drawingFigure 1~8
  • EP2885415B1 patent drawingFigure 2
  • EP2885415B1 patent drawingFigure 3

AI summary

Methods and systems for the production of alcohols are described. A two stage process is utilized, where fermentation in a first stage produces an intermediate product, such as an amino acid or organic acid, from a carbon containing feedstock. A second stage produces alcohol by fermentation of this intermediate product.