Variant Cannabinoid Synthases for High-Purity CBDA Production
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Solution Overview
Problem
Natural cannabinoid synthases from Cannabis sativa plants have limitations in expressing non-abundant cannabinoids, which are difficult to separate in pure forms due to low production levels and heterogeneous product formation, making them costly and suboptimal for therapeutic applications.
Innovation Solution
Development of variant cannabinoid synthases with non-naturally occurring amino acid sequences that alter the yield and ratio of cannabinoids produced, such as CBDA and THCA, by introducing specific mutations at positions like 69, 414, and 445, allowing for increased production of specific cannabinoids in pure forms.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If natural cannabinoid synthases are used to produce non-abundant cannabinoids, then the production process is simple and uses natural enzymes, but the yield is low and product purity is poor due to heterogeneous formation
Solution Approach 1:
The patent applies parameter changes by modifying the amino acid sequence of cannabinoid synthases through site-directed mutagenesis. Specific residues are mutated to alter enzyme specificity and activity, transforming the natural synthase into a variant that produces higher yields of desired cannabinoids with improved purity. This directly addresses the low productivity issue while maintaining the simplicity of enzymatic production.
Solution Approach 2:
The patent applies local quality by making specific localized changes to the enzyme structure through targeted amino acid substitutions at particular positions in the protein sequence. These localized modifications at specific residues alter the enzyme's catalytic properties and substrate specificity, enabling it to produce non-abundant cannabinoids with higher efficiency and purity without changing the overall enzymatic system.
2Ease of manufacture
If natural cannabinoid synthases are used to produce non-abundant cannabinoids, then the production process is simple and uses natural enzymes, but the product purity is poor due to heterogeneous product formation
Solution Approach 1:
The patent applies parameter changes by modifying the amino acid sequence of cannabinoid synthases through site-directed mutagenesis. Specific residues are mutated to alter enzyme specificity and activity, transforming the natural synthase into a variant that produces higher yields of desired cannabinoids with improved purity. This directly addresses the low productivity issue while maintaining the simplicity of enzymatic production.
Solution Approach 2:
The patent applies local quality by making specific localized changes to the enzyme structure through targeted amino acid substitutions at particular positions in the protein sequence. These localized modifications at specific residues alter the enzyme's catalytic properties and substrate specificity, enabling it to produce non-abundant cannabinoids with higher efficiency and purity without changing the overall enzymatic system.
3Productivity
If variant cannabinoid synthases with specific mutations are developed, then the yield and purity of specific cannabinoids are increased, but the enzyme engineering process becomes more complex
Solution Approach 1:
The patent applies parameter changes by modifying the amino acid sequence of cannabinoid synthases through site-directed mutagenesis. Specific residues are mutated to alter enzyme specificity and activity, transforming the natural synthase into a variant that produces higher yields of desired cannabinoids with improved purity. This directly addresses the low productivity issue while maintaining the simplicity of enzymatic production.
Solution Approach 2:
The patent applies local quality by making specific localized changes to the enzyme structure through targeted amino acid substitutions at particular positions in the protein sequence. These localized modifications at specific residues alter the enzyme's catalytic properties and substrate specificity, enabling it to produce non-abundant cannabinoids with higher efficiency and purity without changing the overall enzymatic system.
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 variant synthases achieve higher yields and purer production of cannabinoids like CBDA and THCA, enabling more efficient and cost-effective industrial-scale production of non-abundant cannabinoids for therapeutic use.
Implementation Method 1
The different synthases catalyzing the oxidocyclization of CBGA into THCA, CBDA or CBCA
Implementation Method 2
The different synthases catalyzing the oxidocyclization of CBGA into THCA, CBDA or CBCA
Data Source
AI summary
Described herein are variant, novel cannabinoid synthases, nucleic acids encoding same, and various uses thereof.