Phospholipase C Mutation for Higher Vegetable Oil Degumming Activity
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Solution Overview
Problem
Existing phospholipase C enzymes used in vegetable oil degumming processes have limitations in enzyme activity and efficiency, leading to suboptimal oil yield and increased production costs.
Innovation Solution
Mutating the glycine at amino acid position 10 of BC-PC-PLC to aspartic acid, along with other specific mutations, results in a phospholipase C variant with significantly enhanced specific enzyme activity, improving degumming efficiency and DAG yield.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If phospholipase C enzyme is used in vegetable oil degumming process, then phospholipids can be hydrolyzed to improve oil quality, but the enzyme activity and degumming efficiency are insufficient leading to suboptimal oil yield
Solution Approach 1:
The patent applies parameter changes by mutating specific amino acid residues (Glycine to Aspartic acid at position 10, and other positions) in the phospholipase C enzyme sequence. This chemical parameter change at the molecular level fundamentally alters the enzyme's catalytic properties, increasing specific enzyme activity by 16-fold and improving overall degumming efficiency while maintaining enzyme stability and reliability.
2Ease of manufacture
If conventional phospholipase C is used, then degumming process can be performed, but production costs increase due to suboptimal efficiency requiring more enzyme or repeated processing
Solution Approach 1:
By changing the amino acid parameters of the enzyme through directed evolution and site-directed mutagenesis, the patent creates a hyper-active phospholipase C variant. This parameter change results in dramatically improved catalytic efficiency, allowing the degumming process to achieve higher oil yields while reducing the amount of enzyme required and minimizing production costs.
Solution Approach 2:
The patent replaces the conventional mechanical/chemical refining methods with an optimized enzymatic system. The mutated phospholipase C provides superior catalytic performance compared to traditional chemical degumming agents, achieving better oil yield and quality while reducing the need for additional processing steps and chemical additives, thereby lowering overall production costs.
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 mutated phospholipase C variant achieves an 83% increase in specific enzyme activity without reducing degumming ability per unit enzyme activity, thereby enhancing oil yield and reducing production costs.
Implementation Method 1
PLC can specifically hydrolyze the phosphoglyceride bond at Sn-3 of a phospholipid and generate the corresponding diacylglycerol (DAG) and phosphatidic acids
Implementation Method 2
Phospholipase is an enzyme widely present in animals, plants, and microorganisms that can specifically hydrolyze glycerophospholipids
Data Source
Figure 1~2
Figure 3
AI summary
According to the present disclosure, on the basis of all existing mutations, the tenth glycine of a BC-PC-PLC is mutated into aspartic acid, a specific enzyme activity thereof is 83% higher than that of a sequence before the mutation, and protein expression and degumming activity of unit enzyme activity do not change, so as to further reduce manufacturing costs.