Growth-Coupled 1,4-Butanediol Production via Metabolic Disruption

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Solution Overview

Problem

Current methods for producing 1,4-butanediol rely on petroleum-based feedstocks, which are energy- and capital-intensive, and the isolation of intermediates like succinic acid is costly and requires high temperatures and pressures.

Innovation Solution

Design and production of non-naturally occurring microorganisms with disrupted genes such as adhE, aspA, and IdhA, equipped with an exogenous 1,4-butanediol biosynthetic pathway involving enzymes like 4-hydroxybutanoate dehydrogenase and succinic semialdehyde dehydrogenase, allowing for efficient biosynthesis of 1,4-butanediol.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If conventional petrochemical routes are used for BDO production, then production capacity is sufficient, but energy consumption and capital costs are high

Engineering Contradiction:
ImproveBDO production capacityVSAvoidenergy consumption
Core Design Contradiction:
ProductivityVSUse of energy by moving object

Solution Approach 1:

The patent replaces mechanical/chemical synthesis systems with a biological system. Engineered microorganisms (E. coli, S. cerevisiae, C. glutamicum) use metabolic pathways to convert renewable substrates (glucose, glycerol, starch, cellulose) directly into 1,4-butanediol through enzymatic reactions, eliminating the need for high-energy petrochemical processes involving acetylene, formaldehyde, and catalytic hydrogenation

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

Solution Approach 2:

The patent changes the fundamental parameters of the production system by shifting from petrochemical feedstocks to renewable biological substrates, and from high-temperature/high-pressure chemical reactions to ambient-condition biological fermentation. This transforms the production paradigm while maintaining commercial viability

Inventive Principle:
Principle #35Parameter changes

2Productivity

If succinic acid isolation and catalytic reduction are used, then BDO can be produced, but isolation costs are high and high temperatures and pressures are required

Engineering Contradiction:
ImproveBDO productionVSAvoidisolation and purification cost
Core Design Contradiction:
ProductivityVSEase of manufacture

Solution Approach 1:

The patent extracts and eliminates the problematic intermediate isolation step by engineering complete biosynthetic pathways that produce BDO directly as the final product. The engineered microorganisms convert substrates through metabolic intermediates (including succinic acid in vivo) directly to BDO, which can be recovered through simple fermentation broth processing rather than costly isolation and purification

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

The patent uses engineered microorganisms as living intermediaries that perform the complex multi-step conversion internally. The microorganisms act as biocatalysts that handle the transformation from substrate to BDO through their metabolic machinery, eliminating the need for separate isolation and catalytic reduction steps

Inventive Principle:
Principle #24Intermediary (Mediator)

3Stability of the object's composition

If gene disruptions are introduced for growth-coupled production, then genetic stability is improved, but organism complexity increases

Engineering Contradiction:
Improvegenetic stabilityVSAvoidgenetic engineering complexity
Core Design Contradiction:
Stability of the object's compositionVSDevice complexity

Solution Approach 1:

The patent performs preliminary genetic engineering actions during strain development to establish growth-coupled production. Gene disruptions (such as in adhE, pfl, and other metabolic genes) are introduced in advance to create metabolic dependencies that force the organism to produce BDO for survival, ensuring genetic stability is built into the system's fundamental biology rather than added later

Inventive Principle:
Principle #10Preliminary action

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 production of 1,4-butanediol using renewable resources, reducing energy and capital costs while achieving high yields and genetic stability for continuous bioprocesses.

Implementation Method 1

a 1,4-butanediol (BDO) biosynthetic pathway, said pathway comprising at least one exogenous nucleic acid encoding 4-hydroxybutanoate dehydrogenase, CoA-independent succinic semialdehyde dehydrogenase, succinyl-CoA synthetase, CoA-dependent succinic semialdehyde dehydrogenase, 4-hydroxybutyrate:CoA transferase, glutamate:succinic semialdehyde transaminase, glutamate decarboxylase, CoA-independent aldehyde dehydrogenase, CoA-dependent aldehyde dehydrogenase or alcohol dehydrogenase

Methodology Applied
Scientific EffectEnzymatic reaction: Enzyme

Data Source

PatentEP3447133B1Methods and organisms for the growth-coupled production of 1,4-butanediol
Publication Date: 2023.01.04 GENOMATICA INC
  • EP3447133B1 patent drawingFigure 1
  • EP3447133B1 patent drawingFigure 2
  • EP3447133B1 patent drawingFigure 3

AI summary

The invention provides a non-naturally occurring microorganism comprising one or more gene disruptions, the one or more gene disruptions occurring in genes encoding an enzyme obligatory to coupling 1,4-butanediol production to growth of the microorganism when the gene disruption reduces an activity of the enzyme, whereby the one or more gene disruptions confers stable growth-coupled production of 1,4-butanediol onto the non-naturally occurring microorganism. The microorganism can further comprise a gene encoding an enzyme in a 1,4-butanediol (BDO) biosynthetic pathway. The invention additionally relates to methods of using microorganisms to produce BDO.