skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Precursor-Directed Combinatorial Biosynthesis of Cinnamoyl, Dihydrocinnamoyl, and Benzoyl Anthranilates in Saccharomyces cerevisiae

Abstract

© 2015 Eudes et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Biological synthesis of pharmaceuticals and biochemicals offers an environmentally friendly alternative to conventional chemical synthesis. These alternative methods require the design of metabolic pathways and the identification of enzymes exhibiting adequate activities. Cinnamoyl, dihydrocinnamoyl, and benzoyl anthranilates are natural metabolites which possess beneficial activities for human health, and the search is expanding for novel derivatives that might have enhanced biological activity. For example, biosynthesis in Dianthus caryophyllus is catalyzed by hydroxycinnamoyl/benzoyl-CoA:anthranilate N-hydroxycinnamoyl/ benzoyltransferase (HCBT), which couples hydroxycinnamoyl-CoAs and benzoyl-CoAs to anthranilate. We recently demonstrated the potential of using yeast (Saccharomyces cerevisiae) for the biological production of a few cinnamoyl anthranilates by heterologous co-expression of 4-coumaroyl:CoA ligase from Arabidopsis thaliana (4CL5) and HCBT. Here we report that, by exploiting the substrate flexibility of both 4CL5 and HCBT, we achieved rapid biosynthesis of more than 160 cinnamoyl, dihydrocinnamoyl, and benzoyl anthranilates in yeast upon feeding with both natural and non-natural cinnamates, dihydrocinnamates, benzoates, and anthranilates. Our results demonstrate the use of enzyme promiscuitymore » in biological synthesis to achieve high chemical diversity within a defined class of molecules. This work also points to the potential for the combinatorial biosynthesis of diverse and valuable cinnamoylated, dihydrocinnamoylated, and benzoylated products by using the versatile biological enzyme 4CL5 along with characterized cinnamoyl- CoA- and benzoyl-CoA-utilizing transferases.« less

Authors:
 [1];  [2];  [1];  [1];  [1];  [3];  [1]
  1. Joint BioEnergy Inst., Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
  2. Joint BioEnergy Inst., Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); San Francisco State Univ., CA (United States)
  3. Joint BioEnergy Inst., Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Univ. of California, Berkeley, CA (United States)
Publication Date:
Research Org.:
Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER)
OSTI Identifier:
1256953
Alternate Identifier(s):
OSTI ID: 1378578
Grant/Contract Number:  
AC02- 05CH11231; AC02-05CH11231
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
PLoS ONE
Additional Journal Information:
Journal Volume: 10; Journal Issue: 10; Journal ID: ISSN 1932-6203
Publisher:
Public Library of Science
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Eudes, Aymerick, Teixeira Benites, Veronica, Wang, George, Baidoo, Edward E. K., Lee, Taek Soon, Keasling, Jay D., and Loqué, Dominique. Precursor-Directed Combinatorial Biosynthesis of Cinnamoyl, Dihydrocinnamoyl, and Benzoyl Anthranilates in Saccharomyces cerevisiae. United States: N. p., 2015. Web. doi:10.1371/journal.pone.0138972.
Eudes, Aymerick, Teixeira Benites, Veronica, Wang, George, Baidoo, Edward E. K., Lee, Taek Soon, Keasling, Jay D., & Loqué, Dominique. Precursor-Directed Combinatorial Biosynthesis of Cinnamoyl, Dihydrocinnamoyl, and Benzoyl Anthranilates in Saccharomyces cerevisiae. United States. https://doi.org/10.1371/journal.pone.0138972
Eudes, Aymerick, Teixeira Benites, Veronica, Wang, George, Baidoo, Edward E. K., Lee, Taek Soon, Keasling, Jay D., and Loqué, Dominique. 2015. "Precursor-Directed Combinatorial Biosynthesis of Cinnamoyl, Dihydrocinnamoyl, and Benzoyl Anthranilates in Saccharomyces cerevisiae". United States. https://doi.org/10.1371/journal.pone.0138972. https://www.osti.gov/servlets/purl/1256953.
@article{osti_1256953,
title = {Precursor-Directed Combinatorial Biosynthesis of Cinnamoyl, Dihydrocinnamoyl, and Benzoyl Anthranilates in Saccharomyces cerevisiae},
author = {Eudes, Aymerick and Teixeira Benites, Veronica and Wang, George and Baidoo, Edward E. K. and Lee, Taek Soon and Keasling, Jay D. and Loqué, Dominique},
abstractNote = {© 2015 Eudes et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Biological synthesis of pharmaceuticals and biochemicals offers an environmentally friendly alternative to conventional chemical synthesis. These alternative methods require the design of metabolic pathways and the identification of enzymes exhibiting adequate activities. Cinnamoyl, dihydrocinnamoyl, and benzoyl anthranilates are natural metabolites which possess beneficial activities for human health, and the search is expanding for novel derivatives that might have enhanced biological activity. For example, biosynthesis in Dianthus caryophyllus is catalyzed by hydroxycinnamoyl/benzoyl-CoA:anthranilate N-hydroxycinnamoyl/ benzoyltransferase (HCBT), which couples hydroxycinnamoyl-CoAs and benzoyl-CoAs to anthranilate. We recently demonstrated the potential of using yeast (Saccharomyces cerevisiae) for the biological production of a few cinnamoyl anthranilates by heterologous co-expression of 4-coumaroyl:CoA ligase from Arabidopsis thaliana (4CL5) and HCBT. Here we report that, by exploiting the substrate flexibility of both 4CL5 and HCBT, we achieved rapid biosynthesis of more than 160 cinnamoyl, dihydrocinnamoyl, and benzoyl anthranilates in yeast upon feeding with both natural and non-natural cinnamates, dihydrocinnamates, benzoates, and anthranilates. Our results demonstrate the use of enzyme promiscuity in biological synthesis to achieve high chemical diversity within a defined class of molecules. This work also points to the potential for the combinatorial biosynthesis of diverse and valuable cinnamoylated, dihydrocinnamoylated, and benzoylated products by using the versatile biological enzyme 4CL5 along with characterized cinnamoyl- CoA- and benzoyl-CoA-utilizing transferases.},
doi = {10.1371/journal.pone.0138972},
url = {https://www.osti.gov/biblio/1256953}, journal = {PLoS ONE},
issn = {1932-6203},
number = 10,
volume = 10,
place = {United States},
year = {Fri Oct 02 00:00:00 EDT 2015},
month = {Fri Oct 02 00:00:00 EDT 2015}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record

Citation Metrics:
Cited by: 12 works
Citation information provided by
Web of Science

Figures / Tables:

Fig 1 Fig 1: Structure of cinnamoyl, dihydrocinnamoyl, and benzoyl anthranilates. (A) Cinnamoyl anthranilates. Tranilast: R1 = R2 = R3 = R6 = H, R4 = R5 = OMe. (B) Dihydrocinnamoyl anthranilates. DHavnD: R1 = R2 = R3 = R4 = R6 = H, R5 = OH. (C) Benzoyl anthranilates. Dianthramide Bmore » from D. caryophyllus: R1-6 = H.« less

Save / Share:

Works referenced in this record:

Avena sativa (Oat), A Potential Neutraceutical and Therapeutic Agent: An Overview
journal, January 2013


A Target-Based High Throughput Screen Yields Trypanosoma brucei Hexokinase Small Molecule Inhibitors with Antiparasitic Activity
journal, April 2010


Production of tranilast [N-(3′,4′-dimethoxycinnamoyl)-anthranilic acid] and its analogs in yeast Saccharomyces cerevisiae
journal, October 2010


Acyltransferases in plants: a good time to be BAHD
journal, June 2006


Balsacones D-I, dihydrocinnamoyl flavans from Populus balsamifera buds
journal, April 2014


Benzamidobenzoic acids as potent PqsD inhibitors for the treatment of Pseudomonas aeruginosa infections
journal, April 2014


Dihydroavenanthramide D protects pancreatic β-cells from cytokine and streptozotocin toxicity
journal, September 2009


A Purpose-Synthesised Anti-Fibrotic Agent Attenuates Experimental Kidney Diseases in the Rat
journal, October 2012


N-Benzoyl anthranilic acid derivatives as selective inhibitors of aldo–keto reductase AKR1C3
journal, September 2012


3′,4′-Bis-difluoromethoxycinnamoylanthranilate (FT061): An orally-active antifibrotic agent that reduces albuminuria in a rat model of progressive diabetic nephropathy
journal, December 2013


FT011, a new anti-fibrotic drug, attenuates fibrosis and chronic heart failure in experimental diabetic cardiomyopathy
journal, May 2012


New Cytotoxic Cyclic Peptides and Dianthramide from Dianthus superbus
journal, September 2004


The 4-coumarate:CoA ligase gene family in Arabidopsis thaliana comprises one rare, sinapate-activating and three commonly occurring isoenzymes
journal, February 2004


Potential health benefits of avenanthramides of oats
journal, December 2009


A Novel Fungal Flavin-Dependent Halogenase for Natural Product Biosynthesis
journal, September 2010


A Familiar Ring to It: Biosynthesis of Plant Benzoic Acids
journal, January 2015


Production of hydroxycinnamoyl anthranilates from glucose in Escherichia coli
journal, January 2013


Flavin-dependent halogenases involved in secondary metabolism in bacteria
journal, March 2006


FT23, an orally active antifibrotic compound, attenuates structural and functional abnormalities in an experimental model of diabetic cardiomyopathy
journal, July 2012


Treatment of Cutaneous Sarcoidosis with Tranilast
journal, February 1995


Dermal and transdermal targeting of dihydroavenanthramide D using enhancer molecules and novel microemulsions
journal, August 2009


Dihydroavenanthramide D inhibits human breast cancer cell invasion through suppression of MMP-9 expression
journal, February 2011


Evaluation and optimization of antifibrotic activity of cinnamoyl anthranilates
journal, December 2009


New antibacterial dihydrochalcone derivatives from buds of Populus balsamifera
journal, March 2013


Inhibitory effects of tranilast on expression of transforming growth factor-β isoforms and receptors in injured arteries
journal, April 1998


Interrogating a Hexokinase-Selected Small-Molecule Library for Inhibitors of Plasmodium falciparum Hexokinase
journal, May 2013


Manufacturing Molecules Through Metabolic Engineering
journal, December 2010


Advancing secondary metabolite biosynthesis in yeast with synthetic biology tools
journal, January 2012


A new anti-fibrotic drug attenuates cardiac remodeling and systolic dysfunction following experimental myocardial infarction
journal, September 2013


NMR-Driven Discovery of Benzoylanthranilic Acid Inhibitors of Far Upstream Element Binding Protein Binding to the Human Oncogene c-myc Promoter
journal, September 2004


Halogenated Heterocycles as Pharmaceuticals
book, January 2011


A Novel Fungal Flavin-Dependent Halogenase for Natural Product Biosynthesis
journal, September 2010


Flavin-dependent halogenases involved in secondary metabolism in bacteria
journal, March 2006


Dihydroavenanthramide D protects pancreatic β-cells from cytokine and streptozotocin toxicity
journal, September 2009


Evaluation and optimization of antifibrotic activity of cinnamoyl anthranilates
journal, December 2009


N-Benzoyl anthranilic acid derivatives as selective inhibitors of aldo–keto reductase AKR1C3
journal, September 2012


3′,4′-Bis-difluoromethoxycinnamoylanthranilate (FT061): An orally-active antifibrotic agent that reduces albuminuria in a rat model of progressive diabetic nephropathy
journal, December 2013


Benzamidobenzoic acids as potent PqsD inhibitors for the treatment of Pseudomonas aeruginosa infections
journal, April 2014


Dermal and transdermal targeting of dihydroavenanthramide D using enhancer molecules and novel microemulsions
journal, August 2009


A new anti-fibrotic drug attenuates cardiac remodeling and systolic dysfunction following experimental myocardial infarction
journal, September 2013


Acyltransferases in plants: a good time to be BAHD
journal, June 2006


Balsacones D-I, dihydrocinnamoyl flavans from Populus balsamifera buds
journal, April 2014


Dianthramides A and B, two N-benzoylanthranilic acid derivatives from elicited tissues of Dianthus caryophyllus
journal, August 1984


NMR-Driven Discovery of Benzoylanthranilic Acid Inhibitors of Far Upstream Element Binding Protein Binding to the Human Oncogene c-myc Promoter
journal, September 2004


New Cytotoxic Cyclic Peptides and Dianthramide from Dianthus superbus
journal, September 2004


The 4-coumarate:CoA ligase gene family in Arabidopsis thaliana comprises one rare, sinapate-activating and three commonly occurring isoenzymes
journal, February 2004


Avena sativa (Oat), A Potential Neutraceutical and Therapeutic Agent: An Overview
journal, January 2013


FT011, a new anti-fibrotic drug, attenuates fibrosis and chronic heart failure in experimental diabetic cardiomyopathy
journal, May 2012


A Familiar Ring to It: Biosynthesis of Plant Benzoic Acids
journal, October 2014


Treatment of Cutaneous Sarcoidosis with Tranilast
journal, February 1995


FT23, an orally active antifibrotic compound, attenuates structural and functional abnormalities in an experimental model of diabetic cardiomyopathy
journal, July 2012


Potential health benefits of avenanthramides of oats
journal, December 2009


Manufacturing Molecules Through Metabolic Engineering
journal, December 2010


Metabolic Engineering of the Phenylpropanoid Pathway in Saccharomyces cerevisiae
journal, June 2005


Production of hydroxycinnamoyl anthranilates from glucose in Escherichia coli
journal, January 2013


A Target-Based High Throughput Screen Yields Trypanosoma brucei Hexokinase Small Molecule Inhibitors with Antiparasitic Activity
journal, April 2010


A Purpose-Synthesised Anti-Fibrotic Agent Attenuates Experimental Kidney Diseases in the Rat
journal, October 2012


Recent advances in combinatorial biosynthesis for drug discovery
journal, February 2015


Works referencing / citing this record:

A straightforward TBHP-mediated synthesis of 2-amidobenzoic acids from 2-arylindoles and their antimicrobial activity
journal, January 2019


Exploiting members of the BAHD acyltransferase family to synthesize multiple hydroxycinnamate and benzoate conjugates in yeast
journal, November 2016


Synthesis of avenanthramides using engineered Escherichia coli
journal, March 2018


Metabolic pairing of aerobic and anaerobic production in a one-pot batch cultivation
journal, July 2018


Metabolic pairing of aerobic and anaerobic production in a one-pot batch cultivation
posted_content, February 2018


Engineering a microbial biosynthesis platform for de novo production of tropane alkaloids
journal, August 2019


Exploiting members of the BAHD acyltransferase family to synthesize multiple hydroxycinnamate and benzoate conjugates in yeast
journal, November 2016