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Cannabinoid Biosynthesis Using Noncanonical Cannabinoid Synthases.

Maybelle Kho Go1,2,3, Tingting Zhu1,2,3, Kevin Jie Han Lim1,2,3

  • 1Synthetic Biology for Clinical and Technological Innovation, National University of Singapore, 14 Medical Drive, Singapore 117599, Singapore.

International Journal of Molecular Sciences
|January 21, 2023
PubMed
Summary
This summary is machine-generated.

Researchers discovered novel enzymes that convert cannabigerolic acid (CBGA) into cannabielsoin (CBE). These uncharacterized berberine bridge enzymes (BBEs) offer new pathways for cannabinoid production.

Keywords:
Pichia pastorisberberine bridge enzymecannabielsoincannabigerolic acidcannabinoid biosynthesiscannabinoid synthase

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Area of Science:

  • Biocatalysis and enzyme engineering
  • Natural product biosynthesis
  • Cannabinoid research

Background:

  • Cannabigerolic acid (CBGA) is a precursor to various cannabinoids.
  • Berberine bridge enzyme (BBE) superfamily enzymes are known for catalyzing oxidative cyclizations.
  • Characterization of BBEs outside of the *Cannabis* genus for cannabinoid biosynthesis is limited.

Purpose of the Study:

  • To identify and characterize novel enzymes capable of catalyzing the oxidative cyclization of CBGA.
  • To explore the potential of non-*Cannabis* BBE homologues in cannabinoid biosynthesis.
  • To establish new methods for the discovery and production of natural and unnatural cannabinoids.

Main Methods:

  • Bioinformatic screening of 232 BBE superfamily homologues.
  • Enzymatic assays using CBGA as a substrate.
  • Recombinant expression and purification of candidate enzymes in *Pichia pastoris*.
  • Analysis of reaction products to confirm CBE formation.

Main Results:

  • Four previously uncharacterized BBE homologues were identified that accept CBGA as a substrate.
  • These enzymes catalyze the oxidative cyclization of CBGA to produce cannabielsoin (CBE).
  • This represents the first successful heterologous expression and functional characterization of non-*Cannabis* BBEs for cannabinoid production from CBGA.

Conclusions:

  • Novel BBE enzymes from diverse organisms can efficiently catalyze the conversion of CBGA to CBE.
  • This discovery opens new avenues for the biotechnological production of cannabinoids.
  • The findings facilitate the exploration of a broader range of natural and synthetic cannabinoids.