Keyword search (3,448 papers available)


Global view of the Clostridium thermocellum cellulosome revealed by quantitative proteomic analysis.

Author(s): Gold ND, Martin VJ

J Bacteriol. 2007 Oct;189(19):6787-95 Authors: Gold ND, Martin VJ

Article GUID: 17644599
Proteomic analysis of Clostridium thermocellum ATCC 27405 reveals the upregulation of an alternative transhydrogenase-malate pathway and nitrogen assimilation in cells grown on cellulose.

Author(s): Burton E, Martin VJ

Can J Microbiol. 2012 Dec;58(12):1378-88 Authors: Burton E, Martin VJ

Article GUID: 23210995
Expression of a library of fungal β-glucosidases in Saccharomyces cerevisiae for the development of a biomass fermenting strain.

Author(s): Wilde C, Gold ND, Bawa N, Tambor JH, Mougharbel L, Storms R, Martin VJ

Appl Microbiol Biotechnol. 2012 Aug;95(3):647-59 Authors: Wilde C, Gold ND, Bawa N, Tambor JH, Mougharbel L, Storms R, Martin VJ

Article GUID: 22218767
Effects of synthetic cohesin-containing scaffold protein architecture on binding dockerin-enzyme fusions on the surface of Lactococcus lactis.

Author(s): Wieczorek AS, Martin VJ

Microb Cell Fact. 2012 Dec 15;11:160 Authors: Wieczorek AS, Martin VJ

Article GUID: 23241215
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
Deconstructing the genetic basis of spent sulphite liquor tolerance using deep sequencing of genome-shuffled yeast.

Author(s): Pinel D, Colatriano D, Jiang H, Lee H, Martin VJ

Biotechnol Biofuels. 2015;8:53 Authors: Pinel D, Colatriano D, Jiang H, Lee H, Martin VJ

Article GUID: 25866561
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
Metabolic engineering of a tyrosine-overproducing yeast platform using targeted metabolomics.

Author(s): Gold ND, Gowen CM, Lussier FX, Cautha SC, Mahadevan R, Martin VJ

Microb Cell Fact. 2015 May 28;14:73 Authors: Gold ND, Gowen CM, Lussier FX, Cautha SC, Mahadevan R, Martin VJ

Article GUID: 26016674
Directed evolution of a fungal β-glucosidase in Saccharomyces cerevisiae.

Author(s): Larue K, Melgar M, Martin VJ

Biotechnol Biofuels. 2016;9:52 Authors: Larue K, Melgar M, Martin VJ

Article GUID: 26949413
Engineering of a Nepetalactol-Producing Platform Strain of Saccharomyces cerevisiae for the Production of Plant Seco-Iridoids.

Author(s): Campbell A, Bauchart P, Gold ND, Zhu Y, De Luca V, Martin VJ

ACS Synth Biol. 2016 05 20;5(5):405-14 Authors: Campbell A, Bauchart P, Gold ND, Zhu Y, De Luca V, Martin VJ

Article GUID: 26981892
Seamless site-directed mutagenesis of the Saccharomyces cerevisiae genome using CRISPR-Cas9.

Author(s): Biot-Pelletier D, Martin VJ

J Biol Eng. 2016;10:6 Authors: Biot-Pelletier D, Martin VJ

Article GUID: 27134651
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
Persistence of Escherichia coli in batch and continuous vermicomposting systems.

Author(s): Hénault-Ethier L, Martin VJ, Gélinas Y

Waste Manag. 2016 Oct;56:88-99 Authors: Hénault-Ethier L, Martin VJ, Gélinas Y

Article GUID: 27499290
Title:Metabolic engineering of a tyrosine-overproducing yeast platform using targeted metabolomics.
Authors:Gold ND,  Gowen CM,  Lussier FX,  Cautha SC,  Mahadevan R,  Martin VJ
Link:https://www.ncbi.nlm.nih.gov/pubmed/26016674?dopt=Abstract
DOI:10.1186/s12934-015-0252-2
Category:Microb Cell Fact
PMID:26016674
Dept Affiliation: GENOMICS
1 Department of Biology and Centre for Structural and Functional Genomics, Concordia University, 7141 Sherbrooke West, Montreal, QC, H4B 1R6, Canada. nicholas.gold@concordia.ca.
2 Department of Chemical Engineering and Applied Chemistry, University of Toronto, 200 College Street, Toronto, ON, M5S 3E5, Canada. chris.gowen@utoronto.ca.
3 Department of Biology and Centre for Structural and Functional Genomics, Concordia University, 7141 Sherbrooke West, Montreal, QC, H4B 1R6, Canada. fx.lussier@concordia.ca.
4 Department of Chemical Engineering and Applied Chemistry, University of Toronto, 200 College Street, Toronto, ON, M5S 3E5, Canada. sarat.cautha@utoronto.ca.
5 Department of Chemical Engineering and Applied Chemistry, University of Toronto, 200 College Street, Toronto, ON, M5S 3E5, Canada. krishna.mahadevan@utoronto.ca.
6 Institute of Biomaterials and Biomedical Engineering, University of Toronto, 164 College Street, Toronto, ON, M5S 3G9, Canada. krishna.mahadevan@utoronto.ca.
7 Department of Biology and Centre for Structural and Functional Genomics, Concordia University, 7141 Sherbrooke West, Montreal, QC, H4B 1R6, Canada. vincent.martin@concordia.ca.

Description:

Metabolic engineering of a tyrosine-overproducing yeast platform using targeted metabolomics.

Microb Cell Fact. 2015 May 28;14:73

Authors: Gold ND, Gowen CM, Lussier FX, Cautha SC, Mahadevan R, Martin VJ

Abstract

BACKGROUND: L-tyrosine is a common precursor for a wide range of valuable secondary metabolites, including benzylisoquinoline alkaloids (BIAs) and many polyketides. An industrially tractable yeast strain optimized for production of L-tyrosine could serve as a platform for the development of BIA and polyketide cell factories. This study applied a targeted metabolomics approach to evaluate metabolic engineering strategies to increase the availability of intracellular L-tyrosine in the yeast Saccharomyces cerevisiae CEN.PK. Our engineering strategies combined localized pathway engineering with global engineering of central metabolism, facilitated by genome-scale steady-state modelling.

RESULTS: Addition of a tyrosine feedback resistant version of 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase Aro4 from S. cerevisiae was combined with overexpression of either a tyrosine feedback resistant yeast chorismate mutase Aro7, the native pentafunctional arom protein Aro1, native prephenate dehydrogenase Tyr1 or cyclohexadienyl dehydrogenase TyrC from Zymomonas mobilis. Loss of aromatic carbon was limited by eliminating phenylpyruvate decarboxylase Aro10. The TAL gene from Rhodobacter sphaeroides was used to produce coumarate as a simple test case of a heterologous by-product of tyrosine. Additionally, multiple strategies for engineering global metabolism to promote tyrosine production were evaluated using metabolic modelling. The T21E mutant of pyruvate kinase Cdc19 was hypothesized to slow the conversion of phosphoenolpyruvate to pyruvate and accumulate the former as precursor to the shikimate pathway. The ZWF1 gene coding for glucose-6-phosphate dehydrogenase was deleted to create an NADPH deficiency designed to force the cell to couple its growth to tyrosine production via overexpressed NADP(+)-dependent prephenate dehydrogenase Tyr1. Our engineered Zwf1(-) strain expressing TYRC ARO4(FBR) and grown in the presence of methionine achieved an intracellular L-tyrosine accumulation up to 520 µmol/g DCW or 192 mM in the cytosol, but sustained flux through this pathway was found to depend on the complete elimination of feedback inhibition and degradation pathways.

CONCLUSIONS: Our targeted metabolomics approach confirmed a likely regulatory site at DAHP synthase and identified another possible cofactor limitation at prephenate dehydrogenase. Additionally, the genome-scale metabolic model identified design strategies that have the potential to improve availability of erythrose 4-phosphate for DAHP synthase and cofactor availability for prephenate dehydrogenase. We evaluated these strategies and provide recommendations for further improvement of aromatic amino acid biosynthesis in S. cerevisiae.

PMID: 26016674 [PubMed - indexed for MEDLINE]