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
Engineering 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
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.
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
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.
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
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.
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
Implementation Method 2
a copolymerized PHA containing a 3HH unit is produced by culturing the transformant
Data Source
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.