Copolymerized PHA with 3HH Units via Multifunctional Enzyme

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Solution Overview

Problem

Current methods for producing copolymerized PHA containing 3HH units have limitations in achieving high composition proportions, which restricts the polymer's softness and versatility for applications such as films, sheets, and soft packaging due to inadequate physical property changes.

Innovation Solution

Introducing a gene encoding an enzyme with trans-2-enoyl-CoA hydratase and (R)-3-hydroxyacyl-CoA dehydrogenase activity, typically from eukaryotic sources, into prokaryotic microorganisms with a PHA synthetase gene to enhance the production of copolymerized PHA with higher 3HH units, along with additional genes like crotonyl-CoA reductase and ethylmalonyl-CoA decarboxylase for improved metabolic routes.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Ease of operation

If the molar fraction of 3HV in P(3HB-co-3HV) is increased to improve softness, then softness is enhanced, but physical property changes are insufficient for working into films, sheets, and soft package containers

Engineering Contradiction:
ImprovesoftnessVSAvoidphysical property change range
Core Design Contradiction:
Ease of operationVSAdaptability or versatility

Solution Approach 1:

The patent changes the chemical composition parameter by introducing 3-hydroxyhexanoic acid (3HH) units into the PHA polymer chain, replacing the use of 3-hydroxyvaleric acid (3HV). This parameter change enables broader physical property adjustments, allowing the polymer to be worked into films, sheets, and soft package containers while maintaining softness.

Inventive Principle:
Principle #35Parameter changes

2Ease of operation

If P(3HB-co-3HH) is produced using wild strain of Aeromonas caviae to increase 3HH composition proportion, then softness is heightened, but the production method has limited applicability

Engineering Contradiction:
ImprovesoftnessVSAvoidproduction method applicability
Core Design Contradiction:
Ease of operationVSAdaptability or versatility

Solution Approach 1:

The patent creates a universal production system by introducing a multifunctional enzyme (trans-2-enoyl-CoA hydratase and (R)-3-hydroxyacyl-CoA dehydrogenase) into prokaryotic microorganisms. This enables various prokaryotic strains to produce P(3HB-co-3HH) with high 3HH composition proportion, expanding production method applicability beyond the original wild strain of Aeromonas caviae.

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

3Ease of operation

If a multifunctional enzyme gene is introduced into prokaryotic microorganism to produce copolymerized PHA with higher 3HH units, then softness and physical properties are enhanced, but device complexity increases

Engineering Contradiction:
ImprovesoftnessVSAvoidgene introduction complexity
Core Design Contradiction:
Ease of operationVSDevice complexity

Solution Approach 1:

The patent merges two separate enzyme functions (trans-2-enoyl-CoA hydratase and (R)-3-hydroxyacyl-CoA dehydrogenase) into a single multifunctional enzyme. This consolidation simplifies the genetic engineering process by introducing one gene instead of multiple separate genes, reducing the complexity of gene introduction while still achieving high 3HH units in the copolymerized PHA.

Inventive Principle:
Principle #5Merging (Combining)

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 copolymerized PHA with significantly higher 3HH units, enhancing the polymer's softness and broadening its physical properties, making it suitable for a wider range of applications beyond traditional uses.

Implementation Method 1

a gene encoding an enzyme having trans-2-enoyl-CoA hydratase activity and (R)-3-hydroxyacyl-CoA dehydrogenase activity is introduced into a prokaryotic microorganism having a PHA synthetase gene capable of synthesizing the copolymerized PHA containing the 3HH units

Methodology Applied
Scientific EffectEnzyme catalysis: Enzyme

Implementation Method 2

a copolymerized PHA containing a 3HH unit is produced by culturing the transformant

Methodology Applied
Scientific EffectFermentation: Fermentation

Data Source

PatentUS10829793B2Transformant that produces copolymerized PHA containing 3HH unit, and method for producing said PHA
Publication Date: 2020.11.10 KANEKA CORP

AI summary

Provided are a transformant that produces a copolymerized PHA containing 3HH units in a higher composition proportion; and a method for producing a copolymerized PHA, using this transformant. The transformant is a transformant that produces a copolymerized PHA containing 3HH units, in which a gene encoding an enzyme having trans-2-enoyl-CoA hydratase activity and (R)-3-hydroxyacyl-CoA dehydrogenase activity is introduced into a prokaryotic microorganism having a PHA synthetase gene capable of synthesizing the copolymerized PHA containing the 3HH units. The method is a method for producing a copolymerized PHA containing 3HH units, which includes a step of culturing this transformant.