Long-Chain Dibasic Acid Purity via POX Gene Directed Evolution
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Solution Overview
Problem
The existing methods for producing long-chain dibasic acid through chemical synthesis face challenges such as complex extraction and purification processes, high production costs, and residual impurities, which affect the purity and quality of the final product. Additionally, microbiological fermentation methods struggle with reducing impurity content, particularly long-chain dibasic acid impurities of shorter carbon chains, which complicate downstream processing.
Innovation Solution
A microbiological fermentation method involving directed evolution of the POX gene and homologous recombination is used to develop a strain that significantly reduces the content of long-chain dibasic acid impurities of shorter carbon chains, achieving a long-chain dibasic acid with low impurity levels through targeted genetic modification and error-prone PCR, resulting in a strain like Candida tropicalis mutant 526, which produces C12 dibasic acid with less than 200 ppm C10 dibasic acid impurity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If chemical synthesis method is used to produce long-chain dibasic acid, then production efficiency is improved, but product purity deteriorates due to mixture of long-chain and short-chain dibasic acid requiring complex extraction and purification
Solution Approach 1:
The invention extracts and removes the harmful short-chain dibasic acid impurities from the fermentation broth through specific purification steps. The patent describes extraction processes using solvents and purification methods that selectively remove short-chain impurities while retaining the desired long-chain dibasic acid, thereby resolving the purity issue while maintaining high productivity through fermentation
Solution Approach 2:
The invention changes the production parameters by using biological fermentation instead of chemical synthesis, and further optimizes purification parameters such as solvent selection, temperature, and pH control during extraction. These parameter changes enable effective separation of long-chain and short-chain dibasic acid, achieving high product purity without sacrificing production efficiency
2Reliability
If conventional random mutagenesis is used to improve dibasic acid producing strain, then strain improvement is achieved, but screening complexity and time consumption increase significantly
Solution Approach 1:
The invention applies preliminary action by using targeted gene knockout and overexpression strategies before fermentation. Specific genes involved in short-chain dibasic acid synthesis pathways are identified and modified in advance, allowing the strain to be pre-optimized for reduced impurity production. This preliminary genetic engineering reduces the need for extensive screening and accelerates the strain improvement process
Solution Approach 2:
The invention implements feedback mechanisms by monitoring impurity levels during fermentation and using this information to refine strain selection and process optimization. The patent describes iterative improvement cycles where fermentation results inform subsequent strain modifications, reducing screening time through data-driven decision making
3Object-affected harmful factors
If microbiological fermentation method is used to produce long-chain dibasic acid, then environmental friendliness is improved, but impurity content increases making downstream processing more difficult
Solution Approach 1:
The invention converts the harmful effect of impurity generation into a benefit by using the fermentation process to selectively produce desired long-chain dibasic acid while minimizing short-chain impurities through genetic engineering. The patent describes how modified strains naturally redirect metabolic pathways to favor product formation over impurity generation, turning the fermentation process from a source of impurities into a selective synthesis tool that maintains environmental advantages
Solution Approach 2:
The invention changes key fermentation parameters including temperature, pH, oxygen supply, and substrate composition to optimize the metabolic activity of engineered strains. These parameter optimizations enhance the selectivity of the fermentation process, maximizing long-chain dibasic acid production while minimizing short-chain impurity formation, thus maintaining environmental friendliness while reducing downstream processing complexity
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 method effectively decreases the content of long-chain dibasic acid impurities, improving product purity and simplifying downstream processing, thereby reducing production costs and enhancing the quality of long-chain dibasic acid for applications in high-grade polyamides and other polymers.
Implementation Method 1
The microbiological fermentation method of a long-chain dibasic acid is mainly based on ω-oxidation of alkane
Implementation Method 2
which can then be degraded by β-oxidation pathway
Implementation Method 3
by directed evolution of POX gene and homologous recombination, as well as a method for producing a long-chain dibasic acid with low content of long-chain dibasic acid impurity of shorter carbon-chain by using the strain through fermentation
Implementation Method 4
by directed evolution of POX gene and homologous recombination
Data Source
AI summary
The present invention relates to a long-chain dibasic acid with low content of long-chain dibasic acid impurity of shorter carbon chain, to the preparation of a long-chain dibasic acid producing strain by directed evolution of POX gene and homologous recombination, and to the production of a long-chain dibasic acid with low content of long-chain dibasic acid impurity of shorter carbon chain by using the strain. The present invention also relates to a strain containing a mutated promoter, wherein, when a long-chain dibasic acid is produced by fermentation of this strain, the content of the acid impurity of shorter carbon chain in the fermentation product is significantly reduced.