Reconstituting Plant Secondary Metabolism in Saccharomyces cerevisiae for Production of High-Value Benzylisoquinoline Alkaloids

Benzylisoquinoline alkaloids (BIAs) constitute a diverse class of plant secondary metabolites that includes the opiate analgesics morphine and codeine. Collectively, BIAs exhibit a myriad of pharmacological activities, including antimicrobial, antitussive, antispasmodic, and anticancer properties. D...

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Bibliographic Details
Published in:	Methods in enzymology Vol. 575; p. 195
Main Authors:	Pyne, M E, Narcross, L, Fossati, E, Bourgeois, L, Burton, E, Gold, N D, Martin, V J J
Format:	Journal Article
Language:	English
Published:	United States 2016
Subjects:	Benzylisoquinolines - metabolism Biosynthetic Pathways Genes, Plant Metabolic Engineering - methods Plants - enzymology Plants - genetics Plants - metabolism Saccharomyces cerevisiae - genetics Saccharomyces cerevisiae - metabolism Secondary Metabolism Synthetic Biology - methods Alkaloids Benzylisoquinoline alkaloids Yeast Metabolic engineering Opiates Synthetic biology Saccharomyces cerevisiae Secondary metabolism
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Summary:	Benzylisoquinoline alkaloids (BIAs) constitute a diverse class of plant secondary metabolites that includes the opiate analgesics morphine and codeine. Collectively, BIAs exhibit a myriad of pharmacological activities, including antimicrobial, antitussive, antispasmodic, and anticancer properties. Despite 2500 known BIA products, only a small proportion are currently produced though traditional crop-based manufacturing, as complex stereochemistry renders chemical synthesis of BIAs largely unfeasible. The advent of synthetic biology and sophisticated microbial engineering coupled with recent advances in the elucidation of plant BIA metabolic networks has provided growing motivation for producing high-value BIAs in microbial hosts. Here, we provide a technical basis for reconstituting BIA biosynthetic pathways in the common yeast Saccharomyces cerevisiae. Methodologies outlined in this chapter include fundamental techniques for expressing and assaying BIA biosynthetic enzymes, bioprospecting large libraries of BIA enzyme variants, and reconstituting and optimizing complete BIA formation pathways in yeast. To expedite construction of superior BIA-producing yeast strains, we emphasize high-throughput techniques. Finally, we identify fundamental challenges impeding deployment of yeast-based BIA production platforms and briefly outline future prospects to overcome such barriers.
ISSN:	1557-7988
DOI:	10.1016/bs.mie.2016.02.011