Long-chain dibasic acid production via CYP52A12 mutation
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Solution Overview
Problem
Current methods for producing long-chain dibasic acids through microbiological fermentation face challenges with residual impurities, leading to reduced product purity and increased production costs, necessitating the genetic modification of strains to enhance yield and reduce impurity content.
Innovation Solution
The development of a mutated CYP52A12 gene with specific base mutations is used to create a microorganism strain that reduces the content of monobasic acid impurities during fermentation, achieved through directed evolution and homologous recombination, resulting in a strain capable of producing long-chain dibasic acids with low monobasic acid impurity levels.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If microbiological fermentation method is used to produce long-chain dibasic acid, then product purity is improved and environmental pollution is reduced, but residual impurities remain that affect product quality and increase production cost
Solution Approach 1:
The patent applies parameter changes by mutating the CYP52A12 gene to alter the metabolic parameters of the microorganism. Specifically, base mutations in the gene sequence change the enzyme's catalytic properties, reducing its ability to produce monobasic acid impurities while maintaining dibasic acid production. This genetic parameter modification directly addresses the impurity issue without requiring additional purification steps.
Solution Approach 2:
The patent extracts and removes the harmful monobasic acid impurity by blocking its production pathway through gene mutation. The mutated CYP52A12 gene specifically eliminates the side reaction that produces monobasic acid, effectively extracting the harmful component from the fermentation process output.
2Reliability
If conventional random mutagenesis is used to improve strain, then screening throughput requirement increases and technical limitations arise, but strain improvement can be achieved
Solution Approach 1:
The patent applies preliminary action by performing targeted gene mutation on CYP52A12 before fermentation. Instead of random mutagenesis requiring extensive screening, the specific gene of interest is mutated in advance with a clear functional target, predetermined mutation sites, and expected outcome (reduced monobasic acid production). This preliminary targeted modification eliminates the need for high-throughput screening of random mutants.
Solution Approach 2:
The patent inverts the conventional approach by instead of screening random mutants to find useful traits, it directly modifies the specific gene known to control the unwanted trait (monobasic acid production). This inversion changes the workflow from broad screening to targeted modification, dramatically reducing screening requirements.
3Productivity
If genetic engineering is used to perform targeted genetic modification, then strain yield is improved, but complexity of genetic modification process increases
Solution Approach 1:
The patent applies segmentation by focusing genetic modification on a single specific gene (CYP52A12) rather than attempting comprehensive strain optimization. The complex metabolic pathway is segmented, and only the critical segment (CYP52A12 gene) responsible for monobasic acid production is modified. This segmentation simplifies the overall genetic modification process while achieving the desired yield and purity improvements.
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 modified strain significantly decreases the content of monobasic acid impurities, improving the purity of long-chain dibasic acids and simplifying the extraction and purification processes, thereby reducing production costs and enhancing the quality of the final product.
Implementation Method 1
Microbiological fermentation technology of producing a long-chain dibasic acid
Implementation Method 2
achieved through directed evolution and homologous recombination
Data Source
AI summary
The invention relates to a long-chain dibasic acid with low content of monobasic acid impurity and a production method thereof, in particular to the preparation of a long-chain dibasic acid producing strain by means of directed evolution and homologous recombination, and to the production of a long-chain dibasic acid with low content of monobasic acid impurity by fermentation of said strain. The invention relates to a mutated CYP52A12 gene, homologous gene or variant thereof, which, relative to GenBank Accession Number AY230498 and taking the first base upstream of the start codon ATG as −1, comprises a mutation. The invention relates to a strain comprising said mutated CYP52A12 gene, homologous gene or variant thereof wherein when the strain is fermented to produce a long-chain dibasic acid, the content of monobasic acid impurity in the fermentation product is significantly reduced.