Aspergillus niger fum Gene Knockout for L-Malic Acid Fermentation
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Solution Overview
Problem
Current methods for producing L-malic acid through fermentation, such as chemical synthesis and biological catalysis, face challenges including high costs, environmental concerns, and the formation of heteroacid byproducts, which restrict industrialization due to low product conversion rates and high byproduct content.
Innovation Solution
An Aspergillus niger engineered strain with a knocked-out fumarate hydratase gene is used to reduce byproduct fumaric acid in the fermentation process, improving L-malic acid yield and reducing byproduct accumulation, thereby decreasing downstream purification costs.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If chemical synthesis method is used to produce L-malic acid, then production efficiency is improved, but environmental pollution increases and equipment requirements become more complex
Solution Approach 1:
The patent replaces the chemical synthesis method (which uses high-temperature high-pressure equipment and petroleum-based chemicals) with a biological fermentation method using Aspergillus niger. This substitution eliminates the need for complex high-pressure equipment while maintaining production efficiency, and avoids environmental pollution associated with petroleum-based chemicals and high-temperature processing.
Solution Approach 2:
The patent changes the production parameters from chemical synthesis conditions (high temperature, high pressure) to biological fermentation conditions (moderate temperature, atmospheric pressure). This parameter change simplifies equipment requirements while maintaining productivity and reducing environmental harm.
2Object-affected harmful factors
If biological catalysis method is used to produce L-malic acid, then environmental friendliness is improved, but production cost increases due to enzyme extraction and purification requirements
Solution Approach 1:
The patent extracts and isolates the fumarate hydratase enzyme from Aspergillus niger, then uses this purified enzyme in the fermentation process. By taking out the specific enzyme responsible for fumaric acid production, the patent enables controlled biological catalysis that is environmentally friendly while avoiding the need for complex enzyme extraction and purification processes in the final product.
Solution Approach 2:
The patent uses the Aspergillus niger strain itself to produce L-malic acid through fermentation, eliminating the need for external enzyme addition. The microorganism naturally produces the enzyme fumarate hydratase, which it then uses to catalyze the conversion of fumaric acid to L-malic acid, making the process self-sufficient and cost-effective.
3Object-affected harmful factors
If Aspergillus niger fermentation is used to produce L-malic acid, then environmental friendliness and renewable carbon sources are improved, but byproduct fumaric acid content increases
Solution Approach 1:
The patent converts the harmful byproduct fumaric acid into the desired product L-malic acid by introducing the fumarate hydratase enzyme. This enzyme catalyzes the conversion of fumaric acid to L-malic acid, transforming the harmful byproduct into a valuable product and eliminating the need for additional purification steps.
Solution Approach 2:
The patent changes the metabolic parameters of Aspergillus niger by introducing specific genetic modifications that enhance the activity of fumarate hydratase. This parameter change shifts the metabolic flux toward L-malic acid production while suppressing fumaric acid accumulation, achieving high product yield with minimal byproducts.
4Ease of operation
If conventional Aspergillus niger strain is used for L-malic acid fermentation, then ease of operation is maintained, but product conversion rate remains low
Solution Approach 1:
The patent changes the genetic parameters of the Aspergillus niger strain by introducing mutations or insertions that enhance the activity of key enzymes in the metabolic pathway. This genetic parameter change increases the product conversion rate while maintaining the ease of operation characteristic of Aspergillus niger fermentation, as the modified strain still grows well under standard fermentation conditions.
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 strain increases L-malic acid yield by 1.97% and reduces fumaric acid content by 93.9%, enhancing the efficiency and cost-effectiveness of the fermentation process for industrial production.
Implementation Method 1
a microbiological fermentation method pays more and more attentions because of its environmental friendliness, renewable carbon sources and the like
Implementation Method 2
The biological catalysis method is mainly an immobilized enzyme or immobilized cell transformation method
Implementation Method 3
an Aspergillus niger engineered strain in which a fumarate hydratase gene fum is knocked out
Implementation Method 4
L-malic acid is initially extracted from fruits such as apples, and this method cannot satisfy the demand of a large-scale market
Data Source
AI summary
The disclosure discloses an Aspergillus niger engineered strain for reducing byproduct fumaric acid in a fermentation process of L-malic acid. The Aspergillus niger engineered strain is an Aspergillus niger engineered strain in which a fumarate hydratase gene fum is knocked out. The disclosure overcomes the defects in the prior art, in the current process of producing malic acid through fermentation of Aspergillus niger, byproduct fumaric acid can be accumulated with the generation of malic acid so as to cause the improved cost of the subsequent malic acid purification process. The disclosure provides an Aspergillus niger engineered strain in which a fum gene is knocked out and a method for greatly reducing byproduct fumaric acid in the fermentation production of Aspergillus niger.