TY - JOUR
T1 - Metabolic engineering of yeast for de novo production of kratom monoterpene indole alkaloids
AU - Holtz, Maxence
AU - Rago, Daniela
AU - Nedermark, Ida
AU - Hansson, Frederik G.
AU - Lehka, Beata J.
AU - Hansen, Lea G.
AU - Marcussen, Nils E.J.
AU - Veneman, Wouter J.
AU - Ahonen, Linda
AU - Wungsintaweekul, Juraithip
AU - Acevedo-Rocha, Carlos G.
AU - Dirks, Ron P.
AU - Zhang, Jie
AU - Keasling, Jay D.
AU - Jensen, Michael K.
N1 - Publisher Copyright:
© 2024
PY - 2024
Y1 - 2024
N2 - Monoterpene indole alkaloids (MIAs) from Mitragyna speciosa (“kratom”), such as mitragynine and speciogynine, are promising novel scaffolds for opioid receptor ligands for treatment of pain, addiction, and depression. While kratom leaves have been used for centuries in South-East Asia as stimulant and pain management substance, the biosynthetic pathway of these psychoactives have only recently been partially elucidated. Here, we demonstrate the de novo production of mitragynine and speciogynine in Saccharomyces cerevisiae through the reconstruction of a five-step synthetic pathway from common MIA precursor strictosidine comprising fungal tryptamine 4-monooxygenase to bypass an unknown kratom hydroxylase. Upon optimizing cultivation conditions, a titer of ∼290 μg/L kratom MIAs from glucose was achieved. Untargeted metabolomics analysis of lead production strains led to the identification of numerous shunt products derived from the activity of strictosidine synthase (STR) and dihydrocorynantheine synthase (DCS), highlighting them as candidates for enzyme engineering to further improve kratom MIAs production in yeast. Finally, by feeding fluorinated tryptamine and expressing a human tailoring enzyme, we further demonstrate production of fluorinated and hydroxylated mitragynine derivatives with potential applications in drug discovery campaigns. Altogether, this study introduces a yeast cell factory platform for the biomanufacturing of complex natural and new-to-nature kratom MIAs derivatives with therapeutic potential.
AB - Monoterpene indole alkaloids (MIAs) from Mitragyna speciosa (“kratom”), such as mitragynine and speciogynine, are promising novel scaffolds for opioid receptor ligands for treatment of pain, addiction, and depression. While kratom leaves have been used for centuries in South-East Asia as stimulant and pain management substance, the biosynthetic pathway of these psychoactives have only recently been partially elucidated. Here, we demonstrate the de novo production of mitragynine and speciogynine in Saccharomyces cerevisiae through the reconstruction of a five-step synthetic pathway from common MIA precursor strictosidine comprising fungal tryptamine 4-monooxygenase to bypass an unknown kratom hydroxylase. Upon optimizing cultivation conditions, a titer of ∼290 μg/L kratom MIAs from glucose was achieved. Untargeted metabolomics analysis of lead production strains led to the identification of numerous shunt products derived from the activity of strictosidine synthase (STR) and dihydrocorynantheine synthase (DCS), highlighting them as candidates for enzyme engineering to further improve kratom MIAs production in yeast. Finally, by feeding fluorinated tryptamine and expressing a human tailoring enzyme, we further demonstrate production of fluorinated and hydroxylated mitragynine derivatives with potential applications in drug discovery campaigns. Altogether, this study introduces a yeast cell factory platform for the biomanufacturing of complex natural and new-to-nature kratom MIAs derivatives with therapeutic potential.
KW - Kratom
KW - Metabolic engineering
KW - Mitragyna speciosa
KW - Mitragynine
KW - Monoterpene indole alkaloids
KW - Synthetic biology
U2 - 10.1016/j.ymben.2024.09.011
DO - 10.1016/j.ymben.2024.09.011
M3 - Review
C2 - 39366478
AN - SCOPUS:85205339408
SN - 1096-7176
VL - 86
SP - 135
EP - 146
JO - Metabolic Engineering
JF - Metabolic Engineering
ER -