Engineered E. coli for Renewable 1,4-BDO Production

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Solution Overview

Problem

Current methods for producing 1,4-butanediol and its precursors are energy- and capital-intensive, relying on petroleum-based feedstocks, and alternatives are needed to reduce environmental impact and production costs.

Innovation Solution

Design and production of non-naturally occurring Escherichia coli organisms with a 1,4-butanediol biosynthesis pathway using exogenous nucleic acids encoding specific enzymes to produce 1,4-butanediol and its precursors, such as 4-hydroxybutanoic acid, through metabolic engineering and adaptive evolution.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If petroleum-based feedstocks and catalytic hydrogenation are used to produce 1,4-butanediol, then production efficiency is high, but energy consumption and capital costs are excessive

Engineering Contradiction:
Improveproduction efficiencyVSAvoidenergy consumption
Core Design Contradiction:
ProductivityVSUse of energy by moving object

Solution Approach 1:

The invention changes the fundamental parameters of the production system by replacing petroleum-based feedstocks with renewable biomass feedstocks (glucose, sucrose, cellulose) and replacing high-temperature catalytic hydrogenation with mild enzymatic biocatalysis occurring at physiological temperatures (30-37°C), thereby dramatically reducing energy consumption while maintaining productivity

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The invention substitutes mechanical/chemical processes (catalytic hydrogenation requiring high pressure and temperature) with biological processes (enzymatic reactions in aqueous media at mild conditions), replacing energy-intensive industrial chemistry with sustainable biotechnology

Inventive Principle:
Principle #28Mechanics substitution (Replace mechanical system)

2Ease of manufacture

If petroleum-based feedstocks are used for 1,4-butanediol production, then production costs are lower, but environmental impact increases

Engineering Contradiction:
Improveproduction costVSAvoidenvironmental impact
Core Design Contradiction:
Ease of manufactureVSObject-affected harmful factors

Solution Approach 1:

The invention changes the feedstock parameter from non-renewable petroleum to renewable biomass, transforming the environmental profile of the process while utilizing readily available, low-cost agricultural materials that can be processed through established fermentation technologies

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The invention converts potentially harmful petroleum extraction and processing into beneficial renewable resource utilization, where agricultural byproducts and biomass waste streams are transformed into valuable chemical feedstocks through microbial fermentation, creating both economic and environmental value

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

3Use of energy by moving object

If succinic acid is used as an intermediate for butanediol production, then renewable feedstock is utilized, but isolation and purification costs are high and high temperatures and pressures are required

Engineering Contradiction:
Improverenewable feedstock utilizationVSAvoidisolation and purification cost
Core Design Contradiction:
Use of energy by moving objectVSEase of manufacture

Solution Approach 1:

The invention extracts and eliminates the problematic intermediate succinic acid from the biosynthetic pathway, replacing it with a direct pathway that converts glucose to 1,4-butanediol through engineered microbial metabolism, thereby avoiding the need for costly isolation and purification steps

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

The invention segments the biosynthetic pathway into discrete enzymatic steps catalyzed by specific engineered enzymes (glucose isomerase, aldolase, dehydrogenases), allowing each step to be optimized and controlled independently, avoiding the accumulation of intermediate compounds that would require separation

Inventive Principle:
Principle #1Segmentation

4Device complexity

If conventional chemical synthesis pathways are used, then process simplicity is maintained, but reliance on non-renewable resources continues

Engineering Contradiction:
Improveprocess simplicityVSAvoidrenewable resource utilization
Core Design Contradiction:
Device complexityVSAdaptability or versatility

Solution Approach 1:

The invention creates a universal biocatalytic system using engineered microorganisms that can process multiple renewable feedstocks (glucose, sucrose, cellulose, lignocellulosic biomass) through the same metabolic pathway, providing both process simplicity and adaptability to various sustainable resources

Inventive Principle:
Principle #6Universality (Multi-functionality)

Solution Approach 2:

The invention introduces engineered microorganisms as living intermediaries that bridge renewable biomass feedstocks and 1,4-butanediol product, performing the complex multi-step chemical transformations that would otherwise require multiple separate chemical processing units

Inventive Principle:
Principle #24Intermediary (Mediator)

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 the efficient, renewable production of 1,4-butanediol and its derivatives, reducing the reliance on petroleum-based feedstocks and energy consumption, while maintaining genetic stability for continuous bioprocesses.

Implementation Method 1

containing a 1,4-butanediol (BDO) pathway comprising exogenous nucleic acids encoding BDO pathway enzymes expressed in a sufficient amount to produce BDO

Methodology Applied
Scientific EffectEnzyme catalysis: Enzyme

Data Source

PatentEP2334800B1Microorganisms for the production of 1,4-butanediol
Publication Date: 2018.12.26 GENOMATICA INC
  • EP2334800B1 patent drawingFigure 1
  • EP2334800B1 patent drawingFigure 2
  • EP2334800B1 patent drawingFigure 3(a)~3(c)

AI summary

The invention provides non- naturally occurring microbial organisms comprising a 1,4-butanediol (BDO) pathway comprising at least one exogenous nucleic acid encoding a BDO pathway enzyme expressed in a sufficient amount to produce BDO. The invention additionally provides methods of using such microbial organisms to produce BDO.