Original Research Paper

Biotechnology Letters

, Volume 37, Issue 9, pp 1869-1875

First online:

Production of Δ9-tetrahydrocannabinolic acid from cannabigerolic acid by whole cells of Pichia (Komagataella) pastoris expressing Δ9-tetrahydrocannabinolic acid synthase from Cannabis sativa l.

  • Bastian ZirpelAffiliated withLaboratory of Technical Biochemistry, Department of Biochemical & Chemical Engineering, TU Dortmund University
  • , Felix StehleAffiliated withLaboratory of Technical Biochemistry, Department of Biochemical & Chemical Engineering, TU Dortmund University Email author 
  • , Oliver KayserAffiliated withLaboratory of Technical Biochemistry, Department of Biochemical & Chemical Engineering, TU Dortmund University

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Abstract

Objective

The Δ9-tetrahydrocannabinolic acid synthase (THCAS) from Cannabis sativa was expressed intracellularly in different organisms to investigate the potential of a biotechnological production of Δ9-tetrahydrocannabinolic acid (THCA) using whole cells.

Results

Functional expression of THCAS was obtained in Saccharomyces cerevisiae and Pichia (Komagataella) pastoris using a signal peptide from the vacuolar protease, proteinase A. No functional expression was achieved in Escherichia coli. The highest volumetric activities obtained were 98 pkat ml−1 (intracellular) and 44 pkat ml−1 (extracellular) after 192 h of cultivation at 15 °C using P. pastoris cells. Low solubility of CBGA prevents the THCAS application in aqueous cell-free systems, thus whole cells were used for a bioconversion of cannabigerolic acid (CBGA) to THCA. Finally, 1 mM (0.36 g THCA l−1) THCA could be produced by 10.5 gCDW l−1 before enzyme activity was lost.

Conclusion

Whole cells of P. pastoris offer the capability of synthesizing pharmaceutical THCA production

Keywords

Cannabigerolic acid Cannabis sativa Pichia pastoris Δ9-Tetrahydrocannabinolic acid Synthase Whole cell bioconversion