Genetically Modified Yeast for Squalene Production
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Solution Overview
Problem
Current methods for producing squalene are costly and limited, hindering widespread use in biodegradable lubricants, lubricant additives, and hydraulic fluids due to high production costs and temperature-related drawbacks of existing biodegradable alternatives.
Innovation Solution
Genetically altered yeast strains are developed to increase squalene production by modifying enzymes such as acetyl-CoA carboxylase, HMG-CoA reductase, squalene epoxidase, and squalene synthase through gene repair oligonucleobases, enhancing enzyme activity and expression levels to optimize isoprenoid biosynthesis.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If squalene is produced by extraction from olive oil or shark liver oil, then squalene can be obtained with excellent lubricity and oxidative stability, but the unit cost is high
Solution Approach 1:
The patent uses yeast cells as disposable biocatalysts that can be cultured cheaply on industrial scale. The yeast is grown, harvested, and discarded after squalene extraction, replacing expensive animal-derived sources with inexpensive microbial cells that can be rapidly reproduced.
Solution Approach 2:
The yeast cells naturally produce squalene as part of their metabolic pathway. By genetically modifying the yeast to overproduce squalene and then harvesting it, the system uses the organism's own biosynthetic machinery to generate the desired product, eliminating the need for complex chemical synthesis or expensive extraction from animal sources.
2Adaptability or versatility
If biodegradable lubricants are derived from vegetable and animal fats and oils, then they have good biodegradability, but they solidify at relatively high temperatures and have flash points that are too low
Solution Approach 1:
The patent changes the chemical composition parameters of the lubricant by using squalene (a triterpene hydrocarbon) as the base stock instead of traditional vegetable or animal fats. Squalene has a lower solidification point and higher flash point while maintaining biodegradability, thus optimizing the temperature performance parameters.
Solution Approach 2:
The patent creates a composite lubricant system by combining squalene with squalane (hydrogenated squalene) and various additives. This composite formulation achieves both biodegradability and improved temperature stability, overcoming the limitations of single-component natural oil lubricants.
3Productivity
If genetically altered yeast strains are used to increase squalene production, then production cost decreases and scalability increases, but the complexity of genetic modification processes increases
Solution Approach 1:
The genetic modification process is segmented into discrete, manageable steps: selecting target genes (HMGR, squalene synthase), designing specific mutations, introducing mutations via transformation, and validating squalene production. This segmentation makes the complex process systematic and scalable.
Solution Approach 2:
The patent changes key genetic parameters by mutating specific genes to increase enzyme activity. By modifying gene sequences to enhance HMGR and squalene synthase expression and activity, the yeast naturally produces higher volumes of squalene without requiring complex external intervention during production.
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 genetically modified yeast strains significantly increase squalene production, enabling cost-effective large-scale manufacturing of biodegradable lubricants and hydraulic fluids with improved temperature stability.
Implementation Method 1
modifying enzymes such as acetyl-CoA carboxylase, HMG-CoA reductase, squalene epoxidase, and squalene synthase through gene repair oligonucleobases
Implementation Method 2
enhancing enzyme activity and expression levels to optimize isoprenoid biosynthesis
Implementation Method 3
genetically altered yeast strains are developed to increase squalene production by modifying enzymes... to optimize isoprenoid biosynthesis
Implementation Method 4
Production of Squalene Using Yeast
Data Source
AI summary
Provided herein compositions and methods for producing isoprenoids, including squalene. In certain aspects and embodiments provided are genetically altered yeast and uses therefore. In some aspects and embodiments, the genetically altered yeast produce isoprenoids, preferably squalene. The genetically altered yeast may have alterations in the expression or activity of enzymes involved in squalene production, for example, acetyl-CoA carboxylase (or “ACCase”), HMG-CoA reductase, squalene epoxidase, and squalene synthase. One or more genes of a genetically altered yeast may be modified by gene repair oligonucleobases. Also are provided methods of producing squalene using a genetically altered yeast. The invention also provides squalene produced by genetically altered yeast.