Biotechnology Letters

, Volume 37, Issue 9, pp 1869–1875

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

Authors

  • Bastian Zirpel
    • Laboratory of Technical Biochemistry, Department of Biochemical & Chemical EngineeringTU Dortmund University
    • Laboratory of Technical Biochemistry, Department of Biochemical & Chemical EngineeringTU Dortmund University
  • Oliver Kayser
    • Laboratory of Technical Biochemistry, Department of Biochemical & Chemical EngineeringTU Dortmund University
Original Research Paper

DOI: 10.1007/s10529-015-1853-x

Cite this article as:
Zirpel, B., Stehle, F. & Kayser, O. Biotechnol Lett (2015) 37: 1869. doi:10.1007/s10529-015-1853-x

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

Supplementary material

10529_2015_1853_MOESM1_ESM.docx (928 kb)
Supplementary material 1 (DOCX 928 kb)

Copyright information

© Springer Science+Business Media Dordrecht 2015