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A yeast platform for high-level synthesis of tetrahydroisoquinoline alkaloids.

Author(s): Pyne ME, Kevvai K, Grewal PS, Narcross L, Choi B, Bourgeois L, Dueber JE, Martin VJJ

Nat Commun. 2020 Jul 03;11(1):3337 Authors: Pyne ME, Kevvai K, Grewal PS, Narcross L, Choi B, Bourgeois L, Dueber JE, Martin VJJ

Article GUID: 32620756
An Engineered Aro1 Protein Degradation Approach for Increased cis,cis-Muconic Acid Biosynthesis in Saccharomyces cerevisiae.

Author(s): Pyne ME, Narcross L, Melgar M, Kevvai K, Mookerjee S, Leite GB, Martin VJJ

Appl Environ Microbiol. 2018 Sep 01;84(17): Authors: Pyne ME, Narcross L, Melgar M, Kevvai K, Mookerjee S, Leite GB, Martin VJJ

Article GUID: 29934332
Reconstitution of a 10-gene pathway for synthesis of the plant alkaloid dihydrosanguinarine in Saccharomyces cerevisiae.

Author(s): Fossati E, Ekins A, Narcross L, Zhu Y, Falgueyret JP, Beaudoin GA, Facchini PJ, Martin VJ

Nat Commun. 2014;5:3283 Authors: Fossati E, Ekins A, Narcross L, Zhu Y, Falgueyret JP, Beaudoin GA, Facchini PJ, Martin VJ

Article GUID: 24513861
Synthesis of Morphinan Alkaloids in Saccharomyces cerevisiae.

Author(s): Fossati E, Narcross L, Ekins A, Falgueyret JP, Martin VJ

PLoS One. 2015;10(4):e0124459 Authors: Fossati E, Narcross L, Ekins A, Falgueyret JP, Martin VJ

Article GUID: 25905794
An enzyme-coupled biosensor enables (S)-reticuline production in yeast from glucose.

Author(s): DeLoache WC, Russ ZN, Narcross L, Gonzales AM, Martin VJ, Dueber JE

Nat Chem Biol. 2015 Jul;11(7):465-71 Authors: DeLoache WC, Russ ZN, Narcross L, Gonzales AM, Martin VJ, Dueber JE

Article GUID: 25984720
Microbial Factories for the Production of Benzylisoquinoline Alkaloids.

Author(s): Narcross L, Fossati E, Bourgeois L, Dueber JE, Martin VJJ

Trends Biotechnol. 2016 Mar;34(3):228-241 Authors: Narcross L, Fossati E, Bourgeois L, Dueber JE, Martin VJJ

Article GUID: 26775900
Reconstituting Plant Secondary Metabolism in Saccharomyces cerevisiae for Production of High-Value Benzylisoquinoline Alkaloids.

Author(s): Pyne ME, Narcross L, Fossati E, Bourgeois L, Burton E, Gold ND, Martin VJ

Methods Enzymol. 2016;575:195-224 Authors: Pyne ME, Narcross L, Fossati E, Bourgeois L, Burton E, Gold ND, Martin VJ

Article GUID: 27417930
Mining Enzyme Diversity of Transcriptome Libraries through DNA Synthesis for Benzylisoquinoline Alkaloid Pathway Optimization in Yeast.

Author(s): Narcross L, Bourgeois L, Fossati E, Burton E, Martin VJ

ACS Synth Biol. 2016 12 16;5(12):1505-1518 Authors: Narcross L, Bourgeois L, Fossati E, Burton E, Martin VJ

Article GUID: 27442619
Engineering Plant Secondary Metabolism in Microbial Systems.

Author(s): Pyne ME, Narcross L, Martin VJJ

Plant Physiol. 2019 03;179(3):844-861 Authors: Pyne ME, Narcross L, Martin VJJ PMID: 30643013 [PubMed - indexed for MEDLINE]

Article GUID: 30643013
Title:Mining Enzyme Diversity of Transcriptome Libraries through DNA Synthesis for Benzylisoquinoline Alkaloid Pathway Optimization in Yeast.
Authors:Narcross L,  Bourgeois L,  Fossati E,  Burton E,  Martin VJ
Link:https://www.ncbi.nlm.nih.gov/pubmed/27442619?dopt=Abstract
DOI:10.1021/acssynbio.6b00119
Category:ACS Synth Biol
PMID:27442619
Dept Affiliation: BIOLOGY
1 Department of Biology, Concordia University , Montréal, Québec H4B 1R6, Canada.
2 Centre for Structural and Functional Genomics, Concordia University , Montréal, Québec H4B 1R6, Canada.

Description:

Mining Enzyme Diversity of Transcriptome Libraries through DNA Synthesis for Benzylisoquinoline Alkaloid Pathway Optimization in Yeast.

ACS Synth Biol. 2016 12 16;5(12):1505-1518

Authors: Narcross L, Bourgeois L, Fossati E, Burton E, Martin VJ

Abstract

The ever-increasing quantity of data deposited to GenBank is a valuable resource for mining new enzyme activities. Falling costs of DNA synthesis enables metabolic engineers to take advantage of this resource for identifying superior or novel enzymes for pathway optimization. Previously, we reported synthesis of the benzylisoquinoline alkaloid dihydrosanguinarine in yeast from norlaudanosoline at a molar conversion of 1.5%. Molar conversion could be improved by reduction of the side-product N-methylcheilanthifoline, a key bottleneck in dihydrosanguinarine biosynthesis. Two pathway enzymes, an N-methyltransferase and a cytochrome P450 of the CYP719A subfamily, were implicated in the synthesis of the side-product. Here, we conducted an extensive screen to identify enzyme homologues whose coexpression reduces side-product synthesis. Phylogenetic trees were generated from multiple sources of sequence data to identify a library of candidate enzymes that were purchased codon-optimized and precloned into expression vectors designed to facilitate high-throughput analysis of gene expression as well as activity assay. Simple in vivo assays were sufficient to guide the selection of superior enzyme homologues that ablated the synthesis of the side-product, and improved molar conversion of norlaudanosoline to dihydrosanguinarine to 10%.

PMID: 27442619 [PubMed - indexed for MEDLINE]