De novo production of protoberberine and benzophenanthridine alkaloids through metabolic engineering of yeast.

Jiao X, Fu X, Li Q, Bu J, Liu X, Savolainen O, Huang L, Guo J, Nielsen J, Chen Y

Nat Commun 15 (1) 8759 [2024-10-09; online 2024-10-09]

Protoberberine alkaloids and benzophenanthridine alkaloids (BZDAs) are subgroups of benzylisoquinoline alkaloids (BIAs), which represent a diverse class of plant-specialized natural metabolites with many pharmacological properties. Microbial biosynthesis has been allowed for accessibility and scalable production of high-value BIAs. Here, we engineer Saccharomyces cerevisiae to de novo produce a series of protoberberines and BZDAs, including palmatine, berberine, chelerythrine, sanguinarine and chelirubine. An ER compartmentalization strategy is developed to improve vacuole protein berberine bridge enzyme (BBE) activity, resulting in >200% increase on the production of the key intermediate (S)-scoulerine. Another promiscuous vacuole protein dihydrobenzophenanthridine oxidase (DBOX) has been identified to catalyze two-electron oxidation on various tetrahydroprotoberberines at N7-C8 position and dihydrobenzophenanthridine alkaloids. Furthermore, cytosolically expressed DBOX can alleviate the limitation on BBE. This study highlights the potential of microbial cell factories for the biosynthesis of a diverse group of BIAs through engineering of heterologous plant enzymes.

Integrated Microscopy Technologies Gothenburg [Service]

PubMed 39384562

DOI 10.1038/s41467-024-53045-3

Crossref 10.1038/s41467-024-53045-3

pmc: PMC11464499
pii: 10.1038/s41467-024-53045-3