Engineered Microorganism for PHA Crystallization Speed

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Solution Overview

Problem

Polyhydroxyalkanoic acid (PHA) copolymers, such as PHBH, have slow crystallization speeds, which limits their melt workability and productivity in processes like injection molding and film molding, due to their low crystallization/solidification speed and adhesiveness, making them difficult to work with using conventional machinery.

Innovation Solution

A microorganism, Cupriavidus necator, is engineered to produce a mixture of PHAs with different melting points by incorporating specific PHA synthase genes, allowing for improved crystallization speed and melt workability through controlled culture methods, eliminating the need for separate nucleating agents and reducing production costs.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Strength

If PHBH is used to improve flexibility and reduce brittleness, then the material becomes softer and more flexible, but the crystallization speed becomes very slow

Engineering Contradiction:
ImproveflexibilityVSAvoidcrystallization speed
Core Design Contradiction:
StrengthVSProductivity

Solution Approach 1:

The patent creates a composite material system by co-producing multiple PHA types (PHB, PHBV, PHBH) within the same microorganism. This composite approach allows the material to inherit flexibility from PHBH while gaining rapid crystallization capability from PHB and PHBV components, resolving the contradiction between flexibility and crystallization speed.

Inventive Principle:
Principle #40Composite materials

Solution Approach 2:

The patent introduces spatial heterogeneity in the polymer matrix by having different PHA types crystallize at different rates and locations. The rapidly crystallizing PHB and PHBV form a matrix that provides structural framework, while PHBH provides flexible segments, creating local quality variations that simultaneously achieve flexibility and fast overall crystallization.

Inventive Principle:
Principle #3Local quality

2Temperature

If PHBH is used to reduce melting point and improve melt workability, then the processing temperature is lowered, but the material becomes adhesive and difficult to release from molds

Engineering Contradiction:
Improvemelting pointVSAvoidmold release
Core Design Contradiction:
TemperatureVSEase of operation

Solution Approach 1:

The patent changes the compositional parameters of the PHA mixture by controlling the ratios of different PHA types produced by the engineered microorganism. By adjusting the proportion of high-melting-point PHB and PHBV relative to low-melting-point PHBH, the overall melting behavior and adhesiveness can be tuned to achieve optimal mold release while maintaining low processing temperatures.

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent performs preliminary crystallization of PHB and PHBV components during the cooling process before the PHBH component becomes overly adhesive. This preliminary action creates a partially crystallized structure that reduces adhesiveness and facilitates mold release before the final solidification stage.

Inventive Principle:
Principle #10Preliminary action

3Productivity

If conventional blending methods are used to mix PHB and PHBH, then the nucleating effect can be achieved, but the blending is insufficient and the effect is reduced

Engineering Contradiction:
Improvecrystallization speedVSAvoidblend uniformity
Core Design Contradiction:
ProductivityVSStability of the object's composition

Solution Approach 1:

The patent merges the production of multiple PHA types into a single biological system (the engineered microorganism). This merging ensures that PHB, PHBV, and PHBH are produced simultaneously in controlled ratios and are inherently well-mixed at the molecular level, eliminating the blending uniformity problems associated with conventional mechanical mixing methods.

Inventive Principle:
Principle #5Merging (Combining)

Solution Approach 2:

The engineered microorganism serves itself by simultaneously producing multiple PHA types with different crystallization properties. The cell's metabolic system automatically regulates the production ratios based on available substrates, providing self-regulating compositional control without requiring external blending operations.

Inventive Principle:
Principle #25Self-service

4Productivity

If high-melting-point PHB is used as a nucleating agent for PHBH, then the crystallization speed is improved, but the production cost increases due to separate production and blending requirements

Engineering Contradiction:
Improvecrystallization speedVSAvoidproduction process complexity
Core Design Contradiction:
ProductivityVSDevice complexity

Solution Approach 1:

The engineered microorganism performs multiple functions simultaneously: it produces the base PHBH polymer and also synthesizes the high-melting-point PHB and PHBV components that act as nucleating agents. This multi-functionality eliminates the need for separate nucleating agent production and blending processes, reducing overall process complexity while maintaining fast crystallization speed.

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

Solution Approach 2:

The patent combines the production of polymer matrix (PHBH) and nucleating agents (PHB, PHBV) into a single fermentation process using the engineered microorganism. This merging of previously separate production steps into one integrated biological process simplifies the overall manufacturing流程 and reduces operational complexity.

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

The engineered microorganism produces PHA mixtures with enhanced crystallization speed, improving melt workability and productivity in various manufacturing processes, such as injection molding and film molding, by dispersing PHAs with different melting points at a molecular level, facilitating continuous production and reducing processing temperatures.

Implementation Method 1

a gene encoding a PHA synthase (A) having an amino acid sequence of SEQ ID NO:1 in which the asparagine at position 149 is substituted with serine or the aspartic acid at position 171 is substituted with glycine or both and a gene encoding a PHA synthase (B)

Methodology Applied
Scientific EffectEnzyme catalysis: Enzyme

Implementation Method 2

the resultant PHA mixture can be remarkably improved in crystallization speed

Methodology Applied
Scientific EffectCrystallization: Crystallisation

Implementation Method 3

a method of adding, to the PHBH, a nucleating agent has been attempted

Methodology Applied
Scientific EffectNucleation: Nucleation

Data Source

PatentEP3358002B1Microorganism having PHA synthase-coding genes and method for producing PHA using same
Publication Date: 2024.08.28 KANEKA CORP

AI summary

An object of the invention is to improve the crystallization speed of a PHA copolymer which is to be slowly crystallized, and improve the melt workability and productivity. A microorganism is used which has genes encoding two or more different PHA synthases derived from the genus Aeromonas. The genes encoding the PHA synthases derived from the genus Aeromonas preferably include genes encoding at least two PHA synthases which are capable of synthesizing a copolymer PHA including, as monomer unit species, 3-hydroxybutyric acid and 3-hydroxyhexanoic acid, and which are different in substrate specificity toward 3-hydroxyhexanoic acid from each other. When this microorganism is cultured, a PHA mixture can be produced which includes three or more PHA species different in melting point from each other.