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Rational Design and Modification of NphB for Cannabinoids Biosynthesis
Wenhao Xia1,2, Shimeng Liu2, Huanyu Chu3
1New Cornerstone Science Laboratory, Shaanxi Key Laboratory of Qinling Ecological Intelligent Monitoring and Protection, School of Ecology and Environment, Northwestern Polytechnical University, Xi'an 710072, China.
Molecules (Basel, Switzerland)
|September 28, 2024
Summary
Researchers enhanced prenyltransferase (NphB) enzyme activity and product range for industrial cannabinoid synthesis. Optimized NphB enables direct production of valuable cannabinoids like CBG, improving yield and efficiency for large-scale applications.
Area of Science:
- Biochemistry
- Enzymology
- Cannabinoid Research
Background:
- Cannabinoid research is expanding, particularly in neuropsychopharmacology and mood regulation.
- Prenyltransferase (NphB), crucial for cannabinoid precursor synthesis, requires optimization for industrial applications due to low activity and limited product range.
Purpose of the Study:
- To enhance the catalytic efficiency and product diversity of prenyltransferase (NphB) through rational design and high-throughput screening.
- To enable direct, broad-spectrum production of various cannabinoids, including CBG, CBGA, CBGV, and CBGVA.
Main Methods:
- Rational enzyme design and high-throughput screening were employed to optimize NphB.
- Molecular dynamics simulations were used to investigate the structural basis for enhanced enzyme activity.
Main Results:
- Mutants W3 and W4 showed significant improvements: W3 achieved a 10.6-fold increase in CBG yield and enhanced catalytic efficiency for CBGA and CBGV.
- W4 mutant displayed a 9.3-fold increase in CBGVA activity.
- Structural analysis revealed that active site reconfiguration, disulfide bond formation, and hydrophobic interactions are key to improved NphB performance.
Conclusions:
- Optimized NphB mutants significantly advance cannabinoid synthesis capabilities.
- These findings provide a foundation for the industrial-scale production of diverse cannabinoids, meeting the growing demand.
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