Host Cell Metabolic Engineering for Oxidosqualene Yield
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for producing oxidosqualene, triterpenes, and triterpenoids are economically inefficient due to low yields and high costs associated with extraction and purification, particularly in biotechnological production systems, where enzyme efficiency and post-translational modifications are suboptimal.
Innovation Solution
A heterologous platform is engineered to overexpress specific proteins involved in the mevalonate pathway, including 3-hydroxy-3-methylglutaryl-coenzyme A reductase and other enzymes, combined with targeted gene knockouts and repression of competitive pathways to enhance the production and purification of oxidosqualene, triterpenes, and triterpenoids in host cells like Saccharomyces cerevisiae.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If traditional extraction and purification methods are used for oxidosqualene, triterpenes and triterpenoids, then the production process is simple, but the yield is low and production costs are high
Solution Approach 1:
The patent replaces traditional mechanical/chemical extraction and purification methods with a biotechnological system using genetically engineered host cells. The host cell is engineered to overexpress mevalonate pathway enzymes and oxidosqualene cyclase, converting the extraction process into a biological production system that achieves higher yields through metabolic engineering rather than physical extraction.
Solution Approach 2:
The patent applies parameter changes by modifying the host cell's metabolic parameters through genetic engineering. Specific enzymes in the mevalonate pathway are overexpressed to alter metabolic flux, and the oxidosqualene cyclase is engineered with optimized amino acid sequences to enhance catalytic efficiency, thereby increasing product yield without proportionally increasing process complexity.
2Device complexity
If traditional extraction methods are used, then equipment and process requirements are minimal, but purification costs are high
Solution Approach 1:
The patent replaces complex mechanical purification equipment and multiple chemical purification steps with a biological production system. The engineered host cell naturally produces the target compounds through metabolic pathways, eliminating the need for extensive extraction and purification infrastructure while reducing substance loss through more efficient biological conversion.
3Productivity
If heterologous production systems are used to increase yield, then productivity improves, but enzyme efficiency and post-translational modifications become suboptimal
Solution Approach 1:
The patent applies local quality by selecting a host cell system (such as insect cells or mammalian cells) that provides the specific local cellular environment needed for optimal enzyme function and post-translational modifications. The host cell's native machinery is leveraged to perform critical functions like proper folding, glycosylation, and other modifications that enhance enzyme efficiency, while still achieving high productivity through genetic overexpression.
Solution Approach 2:
The patent uses the host cell's native cellular machinery as an intermediary between the introduced foreign genes and the final product. The host cell provides essential intermediates including proper protein folding environments, post-translational modification enzymes, and quality control mechanisms that ensure high enzyme efficiency, while the introduced genes drive high-level expression of the production pathway.
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 the yield and purity of oxidosqualene, triterpenes, and triterpenoids, reducing production costs and improving metabolic flux, resulting in a high-yield, cost-effective method for manufacturing these compounds.
Implementation Method 1
wherein the host cell is engineered to overexpress a 3-hydroxy-3-methylglutaryl-coenzyme A reductase... and to overexpress a protein comprising at least one amino acid sequence selected from the group consisting of SEQ ID NOs: 2, 3, 4, 5, 6, 7 and 8
Implementation Method 2
Squalene is used—in its oxidized form 2,3-oxidosqualene—to synthesize sterol precursors (e.g. lanosterol in fungi and animals or cycloartenol in plants) or pentacyclic triterpenes via different oxidosqualene cyclases (OSCs)
Data Source
AI summary
The present invention relates to a method of increasing the yield of at least one of oxidosqualene, triterpenes and/or triterpenoids in a specifically engineered host cell and a respective host cell as well as to the use of said host cell for manufacturing the at least one of oxidosqualene, triterpenes and/or triterpenoids.