Botryococcene Synthase Gene Co-Expression in Transgenic Plants
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Solution Overview
Problem
Current methods for producing energy-rich triterpene oils, such as botryococcene, are limited by the slow growth rate of the green algae that synthesizes them, making large-scale production challenging and inefficient.
Innovation Solution
Identification and characterization of the botryococcene synthase gene, which allows for the co-expression with FPP synthase and triterpene methyltransferase genes in transgenic plants, enabling the production of methylated triterpenes through the metabolic diversion of CO2 fixed in photosynthesis.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If green algae cultures are used to produce botryococcene, then triterpene oils can be obtained, but the slow growth rate of the algae limits large-scale production efficiency
Solution Approach 1:
The patent copies the botryococcene synthase gene from Botryococcus braunii algae into transgenic plants, allowing the plants to produce methylated botryococcene and related triterpenes. This copying approach transfers the biosynthetic capability from slow-growing algae to faster-growing plant systems, resolving the contradiction between obtaining triterpene oils and maintaining productivity.
2Quantity of substance
If botryococcene synthase gene is co-expressed with FPP synthase and triterpene methyltransferase genes in transgenic plants, then methylated triterpenes can be produced through metabolic diversion of CO2, but the complexity of genetic engineering increases
Solution Approach 1:
The patent divides the triterpene biosynthesis pathway into three separate gene components: botryococcene synthase gene, FPP synthase gene, and triterpene methyltransferase gene. These segmented genes are co-expressed in transgenic plants to reconstruct the complete biosynthetic pathway, enabling methylated triterpene production while allowing for modular genetic engineering that manages complexity.
Solution Approach 2:
The patent uses FPP (farnesyl diphosphate) as an intermediary molecule in the biosynthetic pathway. FPP serves as the common precursor that is diverted into the triterpene pathway through the action of botryococcene synthase, which converts FPP into methylated botryococcene and related triterpenes. This intermediary approach simplifies the metabolic engineering by providing a clear substrate-product relationship.
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 provides a reliable and cost-effective means to produce renewable biofuels, achieving high yields of valuable sesquiterpenes and facilitating the conversion of these hydrocarbons into fuel suitable for combustion engines.
Implementation Method 1
The botryococcene synthase of this invention is a triterpene synthase enzyme catalyzing the reductive condensation of 2 farnesyl diphosphate (FPP) substrate molecules yielding botryococcene
Implementation Method 2
catalyzing the reductive condensation of 2 farnesyl diphosphate (FPP) substrate molecules yielding botryococcene
Implementation Method 3
The feasibility of this engineering strategy for the production of large amounts of high-valued sesquiterpenes, terpenes consisting of 15 carbons rather than the 30 carbons found in triterpenes, was recently demonstrated... could be derived from the metabolic diversion of CO2 fixed in the process of photosynthesis flowing directly into triterpene biosynthesis and accumulation
Data Source
AI summary
This application relates to the functional identification and characterization of a nucleic acid molecule encoding a triterpene synthase, in particular botryococcene synthase. Also described are host cells comprising the nucleic acid molecules of this invention, proteins encoded by the nucleic acid molecules and methods for using the nucleic acid molecules, transformed hosts and encoded proteins to produce high levels of triterpene hydrocarbons.