Recombinant Acetogenic Cells for 1,4-Butanediol Production

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Solution Overview

Problem

Current methods for producing 1,4-butanediol rely on petroleum-based chemicals or compete with food sources, and there is a lack of efficient technology to convert cellulose into this chemical using microorganisms.

Innovation Solution

A recombinant acetogenic and obligatory anaerobic cell is developed, equipped with specific genes for biosynthesizing 1,4-butanediol from carbon monoxide, carbon dioxide, and cellulose, utilizing pathways like the Wood-Ljungdahl pathway and cellulosome production to efficiently convert these carbon sources into 1,4-butanediol.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Quantity of substance

If conventional production methods using saccharides, glycerol or oil components are used, then 1,4-butanediol can be produced by microorganisms, but the production amounts are limited and there is competition with food sources

Engineering Contradiction:
Improveproduction amount of 1,4-butanediolVSAvoidavailability of carbon sources
Core Design Contradiction:
Quantity of substanceVSAdaptability or versatility

Solution Approach 1:

The patent changes the carbon source parameter from traditional saccharides/glycerol/oil components to syngas (C1 compounds). This parameter change enables unlimited production quantities without competing with food sources, as syngas can be derived from waste, industrial exhaust gas, natural gas or coal

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The recombinant cell is designed to utilize multiple C1 compounds (carbon monoxide, carbon dioxide, formic acid, methanol) as carbon sources. This multi-functionality in carbon source utilization ensures continuous production capability without dependency on limited agricultural resources

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

2Quantity of substance

If syngas is used as carbon source, then unlimited production is possible without food competition, but there is a lack of efficient technology to convert syngas into 1,4-butanediol using microorganisms

Engineering Contradiction:
Improveproduction amount of 1,4-butanediolVSAvoidconversion efficiency of syngas to 1,4-butanediol
Core Design Contradiction:
Quantity of substanceVSEase of manufacture

Solution Approach 1:

The conversion process is segmented into two independent pathways: (1) syngas conversion to intermediate compounds (succinate or α-ketoglutarate) via Wood-Ljungdahl pathway, and (2) intermediate compounds conversion to 1,4-butanediol via introduced enzyme pathways. This segmentation allows optimization of each stage independently, achieving high overall conversion efficiency

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The patent uses intermediate compounds (succinate or α-ketoglutarate) as mediators between syngas and 1,4-butanediol. These intermediates serve as bridging molecules that connect the C1 metabolism pathway with the 1,4-butanediol biosynthesis pathway, enabling efficient conversion through well-established enzymatic reactions

Inventive Principle:
Principle #24Intermediary (Mediator)

3Adaptability or versatility

If cellulose is used as carbon source, then renewable resource utilization is improved, but there is a lack of efficient technology to convert cellulose into 1,4-butanediol

Engineering Contradiction:
Improveutilization of renewable resourcesVSAvoidconversion efficiency of cellulose to 1,4-butanediol
Core Design Contradiction:
Adaptability or versatilityVSEase of manufacture

Solution Approach 1:

The patent uses succinate or α-ketoglutarate as intermediary compounds that bridge cellulose degradation and 1,4-butanediol production. Cellulose is first degraded to glucose, then converted to these intermediates through central metabolism, which finally serve as substrates for 1,4-butanediol biosynthesis. This intermediary approach enables efficient multi-step conversion

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 allows for the production of 1,4-butanediol from renewable sources like syngas and cellulose, reducing reliance on petroleum and minimizing competition with food sources, while enhancing productivity through optimized enzyme expression and tolerance.

Implementation Method 1

utilizing pathways like the Wood-Ljungdahl pathway and cellulosome production to efficiently convert these carbon sources into 1,4-butanediol

Methodology Applied
Scientific EffectWood-Ljungdahl pathway:

Implementation Method 2

a gene encoding CoA-dependent succinate semialdehyde dehydrogenase; a gene encoding 4-hydroxybutyrate dehydrogenase; a gene encoding 4-hydroxybutyryl-CoA transferase; a gene encoding 4-hydroxybutyryl-CoA reductase; and a gene encoding alcohol dehydrogenase

Methodology Applied
Scientific EffectEnzymatic catalysis: Enzyme

Implementation Method 3

utilizing pathways like the Wood-Ljungdahl pathway and cellulosome production to efficiently convert these carbon sources into 1,4-butanediol

Methodology Applied
Scientific EffectCellulosome production:

Data Source

PatentEP3345995B1Recombinant cells, method for producing recombinant cells, and method for producing 1,4-butanediol
Publication Date: 2021.01.06 SEKISUI CHEMICAL CO LTD
  • EP3345995B1 patent drawingFigure 1
  • EP3345995B1 patent drawingFigure 2
  • EP3345995B1 patent drawingFigure 3

AI summary

An object of the present invention is to provide a technique for producing 1,4-butanediol by recombinant cells. Provided is a recombinant cell that is acetogenic and obligatory anaerobic, wherein the recombinant cell includes a gene encoding at least one enzyme selected from the group consisting of succinate semialdehyde dehydrogenase, succinyl-CoA synthase, CoA-dependent succinate semialdehyde dehydrogenase, 4-hydroxybutyrate dehydrogenase, 4-hydroxybutyryl-CoA transferase, 4-hydroxybutyryl-CoA reductase, 4-hydroxybutyraldehyde dehydrogenase, and alcohol dehydrogenase, the gene is expressed in the recombinant cell, and the recombinant cell produces 1,4-butandiol.