Sus scrofa Myoglobin Expression in E. coli via Signal Peptides
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for producing artificial meat struggle with low heme content, leading to an inability to replicate the color of real meat, and the extraction of myoglobin from animal sources is inefficient and costly, limiting large-scale production.
Innovation Solution
The use of an expression vector and an expression engineering bacterium, specifically introducing signal peptides Pel B and Omp A into a recombinant plasmid, to enhance the expression and secretion of Sus scrofa myoglobin (SsMB) in Escherichia coli, increasing protein expression levels by 2.06 and 1.17 times respectively.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If myoglobin is extracted from animal sources, then the color and flavor of artificial meat can be simulated, but the production cost increases and the process is time-consuming
Solution Approach 1:
The patent creates a recombinant expression system that copies the myoglobin gene from animals into E. coli bacteria, allowing industrial-scale production without relying on animal sources. This resolves the contradiction by enabling high-volume production (improving quantity) while maintaining animal-free, efficient production processes (improving productivity).
Solution Approach 2:
The patent replaces the mechanical extraction process from animal tissues with a biological expression system using recombinant DNA technology. This substitution eliminates the need for animal slaughter and manual extraction, thereby improving both production efficiency and scalability while maintaining myoglobin quality.
2Quantity of substance
If myoglobin is extracted from animal muscle tissues, then myoglobin can be obtained, but the supply is limited and spontaneous oxidation occurs
Solution Approach 1:
By copying the myoglobin gene into a stable bacterial expression system, the patent achieves unlimited supply potential through microbial fermentation while maintaining protein stability. The recombinant myoglobin produced in E. coli avoids the spontaneous oxidation issues of animal-derived myoglobin due to controlled production conditions and proper post-translational modification.
3Quantity of substance
If signal peptides Pel B and Omp A are introduced into the recombinant plasmid, then the expression level of SsMB increases, but the plasmid structure becomes more complex
Solution Approach 1:
The patent segments the plasmid structure by inserting distinct signal peptide coding sequences (Pel B and Omp A) as separate functional elements within the recombinant plasmid. This segmentation allows each signal peptide to independently enhance protein expression through specific mechanisms, achieving high expression levels while maintaining manageable plasmid complexity through modular design.
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
This approach significantly improves the yield and expression of SsMB, providing a more efficient and cost-effective method for producing myoglobin, essential for simulating the color and flavor of artificial meat.
Implementation Method 1
The efficient expression and secretion of Sus scrofa myoglobin (SsMB) in Escherichia coli is achieved by introducing two signal peptides, Pel B and Omp A
Implementation Method 2
a signal peptide of pectin lyase Pel B from Erwinia carotovora, a signal peptide of alkaline phosphatase Pho A that regulates metabolism and transport of Escherichia coli, and a signal peptide of outer membrane protein Omp A of the Escherichia coli are inserted into a recombinant plasmid expressing SsMB
Data Source
AI summary
The present disclosure provides myoglobin (MB) and an expression vector and an expression engineering bacterium thereof, and use thereof, and relates to the technical field of genetic engineering. In the present disclosure, recombinant Escherichia coli strains with signal peptides Pel B and Omp A inserted under same conditions have an expression level of Sus scrofa myoglobin (SsMB) increased by 2.06 and 1.17 times, respectively, compared with an original expression strain of the SsMB. The signal peptides that can increase the expression level of the SsMB provide a new idea for research and application of improving the expression level of the SsMB.