Recombinant E. coli Lycopene Biosynthesis via Gene Segmentation
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Solution Overview
Problem
Current methods for producing lycopene using microorganisms result in low yields, hindering the development of economical production processes, despite the growing demand for this antioxidant and anticancer compound.
Innovation Solution
A method involving the transformation of E. coli with a recombinant vector containing new genes such as crtE, crtB, and crtI, isolated from a sea metagenome, along with genes involved in the mevalonate pathway, to enhance lycopene biosynthesis and productivity through fermentation under optimized conditions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If conventional microorganism cultivation methods are used for lycopene production, then the production process is simple, but the yield is low
Solution Approach 1:
The patent divides the lycopene biosynthesis pathway into multiple gene segments (crtE, crtB, crtI) and introduces them separately into E. coli through recombinant vectors, allowing each gene to be optimized and expressed independently to maximize lycopene yield
Solution Approach 2:
The patent creates a composite genetic system by combining multiple heterologous genes (crtE from Erwinia uredovora, crtB from Drosophila melanogaster, crtI from Erwinia uredovora) with E. coli native genes to form a complete lycopene biosynthesis pathway that does not exist in the host organism naturally
2Productivity
If traditional extraction from natural sources is used, then the production method is straightforward, but the productivity is low
Solution Approach 1:
The patent enables E. coli to self-produce lycopene by introducing the complete biosynthesis pathway genes, eliminating the need for extraction from natural sources and allowing the organism to serve itself in producing the desired compound through its own metabolic pathways
Solution Approach 2:
The patent optimizes multiple parameters including gene expression levels, cultivation conditions (temperature, pH, aeration), and media composition to maximize lycopene production rate, transforming the production process from low-yield extraction to high-yield fermentation
3Quantity of substance
If existing recombinant methods are used, then some lycopene can be produced, but the productivity is insufficient for economical production
Solution Approach 1:
The patent performs preliminary optimization of gene sequences, promoter regions, and ribosome binding sites before fermentation to ensure maximum expression efficiency, and conducts strain selection and media optimization in advance to reduce production time while maximizing lycopene concentration
Solution Approach 2:
The patent implements continuous fermentation processes and optimized cultivation protocols that maintain high lycopene production rates throughout the fermentation period, preventing downtime and ensuring continuous accumulation of the product
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 increases lycopene productivity, achieving higher yields in a shorter time compared to previous methods, making it feasible for large-scale, economical production.
Implementation Method 1
transforming the recombinant vector into E. coli
Implementation Method 2
culturing the E. coli transformant and recovering lycopene from the culture medium
Data Source
AI summary
A method of producing lycopene, with high productivity by means of a recombinant bacterial strain includes preparing the recombinant vector containing genes encoding proteins, which are required for lycopene biosynthesis. The genes involved in lycopene biosynthesis are crtE with the nucleotide SEQ ID NO: 8, crtB with the nucleotide SEQ ID NO: 3 and crtI with the nucleotide SEQ ID NO: 5 of the Sequence List. The said recombinant vector is transformed into Escherichia coli (hereafter E. coli). The E. coli transformant is cultured to recover lycopene from the culture medium.