Recombinant E. coli LegH Expression Vectors
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Solution Overview
Problem
Current methods for producing artificial meat struggle to replicate the color and flavor of real meat due to low heme content, leading to high production costs and environmental concerns, particularly in the extraction of leghemoglobin from soybeans, which has limited yield and complex extraction processes.
Innovation Solution
The use of recombinant expression vectors and signal peptides such as Pel B, Pho A, and Omp A in Escherichia coli to enhance the expression and secretion of leghemoglobin, increasing protein yield by 3.01, 1.25, and 1.22 times respectively, facilitating more efficient biosynthesis and simulation of artificial meat.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If leghemoglobin is extracted from soybeans, then heme content is obtained, but the extraction process is complex and production cost is extremely high
Solution Approach 1:
The patent creates a copy of the leghemoglobin production system by transferring the LegH gene into E. coli bacteria through recombinant DNA technology. Instead of extracting Hb from soybeans, the bacteria are engineered to produce Hb autonomously, simplifying the production process from complex extraction to straightforward microbial fermentation.
Solution Approach 2:
The patent replaces the mechanical extraction process from soybeans with a biological production system. By using recombinant bacteria to synthesize Hb, the complex mechanical and chemical extraction operations are substituted with a biological fermentation process that is easier to scale and control.
2Quantity of substance
If leghemoglobin is extracted from soybeans, then heme content is obtained, but production cost is extremely high
Solution Approach 1:
The patent uses E. coli bacteria as temporary, easily replaceable production vessels. These bacteria can be rapidly cultured and discarded after use, replacing the expensive and time-consuming soybean cultivation and extraction system. The bacteria serve as disposable factories that can be quickly regenerated through simple fermentation.
Solution Approach 2:
The patent changes the production parameters from plant-based extraction to microorganism-based synthesis. By shifting from soybean cultivation (long cycle, low yield) to bacterial fermentation (short cycle, high yield), the production cost and time parameters are dramatically improved while maintaining heme content quality.
3Quantity of substance
If traditional livestock and poultry breeding methods are used, then meat production is achieved, but the supply relationship is imbalanced and environmental concerns arise
Solution Approach 1:
The patent replaces traditional animal-based meat production with a bio-engineered system that produces meat components (heme) through bacterial fermentation. This substitutes the complex ecological systems of livestock breeding with a controlled biotechnological process, eliminating environmental harms associated with animal agriculture.
Solution Approach 2:
The patent extracts the essential meat-coloring and meaty-flavor component (heme) from its traditional animal and plant sources, and produces it independently through recombinant bacteria. This separates the desired functional component from the environmentally harmful production systems, allowing meat production without livestock or poultry.
4Ease of manufacture
If artificial meat products are produced without sufficient heme content, then production cost is reduced, but color similar to real meat cannot be achieved
Solution Approach 1:
The patent optimizes the heme concentration parameter in artificial meat by using bacterial fermentation to achieve precise control over Hb content. This allows production of sufficient heme content for proper color development while maintaining cost-effectiveness through efficient bacterial production rather than expensive plant extraction.
Data Source
AI summary
The present disclosure provides a gene expression cassette encoding hemoglobin (Hb) and use thereof, and relates to the technical field of genetic engineering. In the present disclosure, a recombinant Escherichia coli strain with signal peptides Pel B, Pho A, and Omp A inserted under same conditions has an expression level of leghemoglobin (LegH) increased by 3.01, 1.25, and 1.22 times, respectively, compared with an original LegH expressing strain. The present disclosure provides signal peptides that can increase the expression level of the LegH, and provide a new idea for research and application of improving the expression level of the LegH.