Microbial 1,3-Butanediol Production via Enzyme Pathway Engineering

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Solution Overview

Problem

Current methods for producing 1,3-butanediol rely on petroleum-based feedstocks, necessitating the development of renewable feedstock-based routes to reduce environmental impact and economic costs.

Innovation Solution

Engineering non-naturally occurring microbial organisms with specific genetic modifications to express enzymes involved in the 1,3-butanediol pathway, allowing for the production of 1,3-butanediol through fermentation, utilizing exogenous nucleic acids encoding ketone-reducing acetoacetyl-CoA reductase, aldehyde-forming 3-hydroxybutyryl-CoA reductase, and 3-hydroxybutyraldehyde reductase enzymes.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Ease of manufacture

If petroleum-based feedstocks (acetylene or ethylene) are used for 1,3-butanediol production, then the production process is well-established and efficient, but environmental impact increases and economic costs rise due to petroleum price volatility

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

Solution Approach 1:

The patent changes the fundamental parameter of feedstock source from petroleum-based (acetylene/ethylene) to renewable-based (glucose or other sugars), transforming the chemical pathway while maintaining production efficiency. This parameter change resolves the contradiction by eliminating environmental harm associated with petroleum extraction and processing while preserving manufacturability through established fermentation technologies.

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent replaces the chemical synthesis mechanism (petroleum refining and chemical conversion) with a biological mechanism (microbial fermentation). By substituting the mechanical/chemical system with a biological system, the harmful environmental factors of petroleum processing are eliminated while maintaining an efficient, well-established production process through controlled fermentation.

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

2Productivity

If petroleum-based feedstocks are used, then production efficiency is maintained, but economic costs increase due to dependence on volatile petroleum prices

Engineering Contradiction:
Improveproduction efficiencyVSAvoideconomic cost
Core Design Contradiction:
ProductivityVSQuantity of substance

Solution Approach 1:

The patent changes the economic parameter by substituting petroleum-based feedstocks with renewable sugars at more stable prices. This parameter change maintains productivity through efficient fermentation pathways while reducing economic costs by eliminating dependence on volatile petroleum markets, thereby resolving the contradiction between production efficiency and economic cost.

Inventive Principle:
Principle #35Parameter changes

3Ease of manufacture

If traditional chemical synthesis pathways are used, then the process is straightforward and well-established, but renewable feedstock utilization is not achieved

Engineering Contradiction:
Improveprocess simplicityVSAvoidrenewable feedstock utilization
Core Design Contradiction:
Ease of manufactureVSAdaptability or versatility

Solution Approach 1:

The patent applies universality by designing a fermentation pathway that can utilize multiple renewable feedstocks (glucose, fructose, sucrose, and other sugars) through the same microbial system. This multi-functionality resolves the contradiction by maintaining process simplicity while achieving versatility in renewable feedstock utilization, allowing the same well-established fermentation process to handle various sugar sources.

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

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 1,3-butanediol from renewable sources, reducing dependence on petroleum-based feedstocks and providing a sustainable method for producing this valuable chemical.

Implementation Method 1

a ketone-reducing acetoacetyl-CoA reductase

Methodology Applied
Scientific EffectEnzyme catalysis: Enzyme

Implementation Method 2

ketone-reducing acetoacetyl-CoA reductase

Methodology Applied
Scientific EffectReduction reaction: Reduction

Implementation Method 3

an aldehyde forming 3-hydroxybutyryl-CoA reductase

Methodology Applied
Scientific EffectEnzyme catalysis: Enzyme

Implementation Method 4

aldehyde forming 3-hydroxybutyryl-CoA reductase

Methodology Applied
Scientific EffectReduction reaction: Reduction

Implementation Method 5

a 3-hydroxybutyraldehyde reductase

Methodology Applied
Scientific EffectEnzyme catalysis: Enzyme

Implementation Method 6

3-hydroxybutyraldehyde reductase

Methodology Applied
Scientific EffectReduction reaction: Reduction

Implementation Method 7

producing the commodity chemical 1,3-butanediol... through fermentation

Methodology Applied
Scientific EffectFermentation: Fermentation

Implementation Method 8

said non-naturally occurring microbial organism is in a substantially anaerobic culture medium

Methodology Applied
Scientific EffectAnaerobic metabolism: Anaerobic Digestion

Data Source

PatentEP3865569B1Organisms for the production of 1,3-butanediol
Publication Date: 2023.10.04 GENOMATICA INC
  • EP3865569B1 patent drawingFigure 1
  • EP3865569B1 patent drawingFigure 2
  • EP3865569B1 patent drawingFigure 3

AI summary

A non-naturally occurring microbial organism includes a microbial organism having a 1,3-butanediol (1,3-BDO) pathway having at least one exogenous nucleic acid encoding a 1,3-BDO pathway enzyme expressed in a sufficient account to produce 1,3-BDO. The pathway includes an enzyme selected from a 2-amino-4-ketopentanoate (AKP) thiolase, an AKP dehydrogenase, a 2-amino-4-hydroxypentanoate aminotransferase, a 2-amino-4-hydroxypentanoate oxidoreductase (deaminating), a 2-oxo-4-hydroxypentanoate decarboxylase, a 3-hydroxybutyraldehyde reductase, an AKP aminotransferase, an AKP oxidoreductase (deaminating), a 2,4-dioxopentanoate decarboxylase, a 3-oxobutyraldehyde reductase (ketone reducing), a 3-oxobutyraldehyde reductase (aldehyde reducing), a 4-hydroxy-2-butanone reductase, an AKP decarboxylase, a 4-aminobutan-2-one aminotransferase, a 4-aminobutan-2-one oxidoreductase (deaminating), a 4-aminobutan-2-one ammonia-lyase, a butenone hydratase, an AKP ammonia-lyase, an acetylacrylate decarboxylase, an acetoacelyl-CoA reductase (CoA-dependent, aldehyde forming), an acetoacetyl-CoA reductase (CoA-dependent, alcohol forming), an acetoacetyl-CoA reductase (ketone reducing), a 3-hydroxybutyryl-CoA reductase (aldehyde forming), a 3-hydroxybutyryl-CoA reductase (alcohol forming), a 4-hydroxybutyryl-CoA dehydratase, and a crotonase. A method for producing 1,3-BDO, includes culturing such microbial organisms under conditions and for a sufficient period of time to produce 1,3-BDO.