Engineered Enzymes for Cannabinoid Biosynthesis
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Solution Overview
Problem
The production of specific phytocannabinoids like THC, CBD, and CBC is limited by the availability of precursor molecules and geranyl pyrophosphate in the cannabinoid biosynthesis pathway, hindering mass production and the demand for these compounds in the medical and recreational markets.
Innovation Solution
Engineering enzymes involved in the biosynthesis of olivetolic acid (OA) and geranyl pyrophosphate (GPP) to increase the flux of precursors, specifically through optimizing hexanoic acid pathways and using polyketide synthases and cyclases to enhance the production of cannabinoid precursors like CBGA, thereby increasing the titers of THC, CBD, and CBC.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If traditional plant-based cannabinoid production is used, then natural biosynthesis occurs, but production quantity and rate are limited by precursor availability
Solution Approach 1:
The patent modifies enzyme parameters through directed evolution and rational design to enhance catalytic activity and substrate affinity. Specific amino acid mutations in CBGA synthase and other pathway enzymes increase reaction rates and precursor conversion efficiency, directly addressing the limitation of production rate while maintaining precursor availability
Solution Approach 2:
The patent divides the cannabinoid biosynthesis pathway into discrete enzymatic steps and independently optimizes each enzyme. By segmenting the pathway and expressing engineered enzymes separately in heterologous hosts, the system can control flux through each step individually, overcoming the bottleneck of precursor availability that limits traditional whole-plant production
2Productivity
If enzyme engineering is implemented to increase precursor flux, then cannabinoid production capacity is accelerated, but system complexity increases
Solution Approach 1:
The patent employs universal enzyme engineering strategies and standardized genetic toolkits that can be applied across multiple cannabinoid pathway enzymes. The directed evolution platform and protein design methods developed are transferable to optimize different enzymes in the pathway, reducing overall system complexity through methodological universality while achieving high productivity
Solution Approach 2:
The patent uses heterologous host organisms as intermediaries to simplify the production system. By expressing engineered cannabinoid pathway enzymes in model organisms with well-characterized genetics and metabolism, the complexity of working with native plant systems is reduced, while the engineered enzymes mediate the production of high-value cannabinoids with controlled precursor flux
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
Significantly enhances the rate and total quantity of cannabinoid precursor biosynthesis, addressing production limitations and meeting market demands by optimizing enzyme activity and precursor availability in non-native hosts.
Implementation Method 1
The optimization of OA biosynthesis from hexanoic acid is described herein. These methods were also used to produce a large number of OA analogs, including DVA, as shown in FIG. 2. Further, novel enzymes for OA synthesis (and its analogs) were identified and/or improved by engineering.
Implementation Method 2
Engineering enzymes involved in the biosynthesis of olivetolic acid (OA) and geranyl pyrophosphate (GPP) to increase the flux of precursors, specifically through optimizing hexanoic acid pathways and using polyketide synthases and cyclases to enhance the production of cannabinoid precursors like CBGA
Data Source
AI summary
Described herein are the discovery and/or optimization of enzymes involved in the biosynthesis of olivetolic acid, divarinic acid, and analogs thereof.