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Payne JT, Valentic TR, Smolke CD. Complete biosynthesis of the bisbenzylisoquinoline alkaloids guattegaumerine and berbamunine in yeast. Proc Natl Acad Sci U S A. 2021;118(51)doi: 10.1073/pnas.2112520118.
Payne, J. T., Valentic, T. R., & Smolke, C. D. (2021). Complete biosynthesis of the bisbenzylisoquinoline alkaloids guattegaumerine and berbamunine in yeast. Proceedings of the National Academy of Sciences of the United States of America, 118(51), . https://doi.org/10.1073/pnas.2112520118
Payne, James T, et al. "Complete biosynthesis of the bisbenzylisoquinoline alkaloids guattegaumerine and berbamunine in yeast." Proceedings of the National Academy of Sciences of the United States of America vol. 118,51 (2021). doi: https://doi.org/10.1073/pnas.2112520118
Payne JT, Valentic TR, Smolke CD. Complete biosynthesis of the bisbenzylisoquinoline alkaloids guattegaumerine and berbamunine in yeast. Proc Natl Acad Sci U S A. 2021 Dec 21;118(51). doi: 10.1073/pnas.2112520118. PMID: 34903659.
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Proc Natl Acad Sci U S A. 2021 Dec 21;118(51). doi: 10.1073/pnas.2112520118.

Complete biosynthesis of the bisbenzylisoquinoline alkaloids guattegaumerine and berbamunine in yeast.

Proceedings of the National Academy of Sciences of the United States of America

James T Payne, Timothy R Valentic, Christina D Smolke

Affiliations

  1. Department of Bioengineering, Stanford University, Stanford, CA 94305.
  2. Department of Bioengineering, Stanford University, Stanford, CA 94305; [email protected].
  3. Chan Zuckerberg Biohub, San Francisco, CA 94158.

PMID: 34903659 DOI: 10.1073/pnas.2112520118

Abstract

Benzylisoquinoline alkaloids (BIAs) are a diverse class of medicinal plant natural products. Nearly 500 dimeric bisbenzylisoquinoline alkaloids (bisBIAs), produced by the coupling of two BIA monomers, have been characterized and display a range of pharmacological properties, including anti-inflammatory, antitumor, and antiarrhythmic activities. In recent years, microbial platforms have been engineered to produce several classes of BIAs, which are rare or difficult to obtain from natural plant hosts, including protoberberines, morphinans, and phthalideisoquinolines. However, the heterologous biosyntheses of bisBIAs have thus far been largely unexplored. Here, we describe the engineering of yeast strains that produce the Type I bisBIAs guattegaumerine and berbamunine de novo. Through strain engineering, protein engineering, and optimization of growth conditions, a 10,000-fold improvement in the production of guattegaumerine, the major bisBIA pathway product, was observed. By replacing the cytochrome P450 used in the final coupling reaction with a chimeric variant, the product profile was inverted to instead produce solely berbamunine. Our highest titer engineered yeast strains produced 108 and 25 mg/L of guattegaumerine and berbamunine, respectively. Finally, the inclusion of two additional putative BIA biosynthesis enzymes, SiCNMT2 and NnOMT5, into our bisBIA biosynthetic strains enabled the production of two derivatives of bisBIA pathway intermediates de novo: magnocurarine and armepavine. The de novo heterologous biosyntheses of bisBIAs presented here provide the foundation for the production of additional medicinal bisBIAs in yeast.

Copyright © 2021 the Author(s). Published by PNAS.

Keywords: benzylisoquinoline alkaloids; bisbenzylisoquinoline alkaloids; metabolic engineering; plant natural products; protein engineering

Conflict of interest statement

Competing interest statement: C.D.S. is an inventor on a pending patent application and a founder and the CEO of Antheia, Inc.

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