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Rational and combinatorial approaches to engineering styrene production by Saccharomyces cerevisiae
BACKGROUND: Styrene is an important building-block petrochemical and monomer used to produce numerous plastics. Whereas styrene bioproduction by Escherichia coli was previously reported, the long-term potential of this approach will ultimately rely on the use of hosts with improved industrial phenot...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4145238/ https://www.ncbi.nlm.nih.gov/pubmed/25162943 http://dx.doi.org/10.1186/s12934-014-0123-2 |
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author | McKenna, Rebekah Thompson, Brian Pugh, Shawn Nielsen, David R |
author_facet | McKenna, Rebekah Thompson, Brian Pugh, Shawn Nielsen, David R |
author_sort | McKenna, Rebekah |
collection | PubMed |
description | BACKGROUND: Styrene is an important building-block petrochemical and monomer used to produce numerous plastics. Whereas styrene bioproduction by Escherichia coli was previously reported, the long-term potential of this approach will ultimately rely on the use of hosts with improved industrial phenotypes, such as the yeast Saccharomyces cerevisiae. RESULTS: Classical metabolic evolution was first applied to isolate a mutant capable of phenylalanine over-production to 357 mg/L. Transcription analysis revealed up-regulation of several phenylalanine biosynthesis pathway genes including ARO3, encoding the bottleneck enzyme DAHP synthase. To catalyze the first pathway step, phenylalanine ammonia lyase encoded by PAL2 from A. thaliana was constitutively expressed from a high copy plasmid. The final pathway step, phenylacrylate decarboxylase, was catalyzed by the native FDC1. Expression of FDC1 was naturally induced by trans-cinnamate, the pathway intermediate and its substrate, at levels sufficient for ensuring flux through the pathway. Deletion of ARO10 to eliminate the competing Ehrlich pathway and expression of a feedback-resistant DAHP synthase encoded by ARO4(K229L) preserved and promoted the endogenous availability precursor phenylalanine, leading to improved pathway flux and styrene production. These systematic improvements allowed styrene titers to ultimately reach 29 mg/L at a glucose yield of 1.44 mg/g, a 60% improvement over the initial strain. CONCLUSIONS: The potential of S. cerevisiae as a host for renewable styrene production has been demonstrated. Significant strain improvements, however, will ultimately be needed to achieve economical production levels. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12934-014-0123-2) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-4145238 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-41452382014-08-28 Rational and combinatorial approaches to engineering styrene production by Saccharomyces cerevisiae McKenna, Rebekah Thompson, Brian Pugh, Shawn Nielsen, David R Microb Cell Fact Research BACKGROUND: Styrene is an important building-block petrochemical and monomer used to produce numerous plastics. Whereas styrene bioproduction by Escherichia coli was previously reported, the long-term potential of this approach will ultimately rely on the use of hosts with improved industrial phenotypes, such as the yeast Saccharomyces cerevisiae. RESULTS: Classical metabolic evolution was first applied to isolate a mutant capable of phenylalanine over-production to 357 mg/L. Transcription analysis revealed up-regulation of several phenylalanine biosynthesis pathway genes including ARO3, encoding the bottleneck enzyme DAHP synthase. To catalyze the first pathway step, phenylalanine ammonia lyase encoded by PAL2 from A. thaliana was constitutively expressed from a high copy plasmid. The final pathway step, phenylacrylate decarboxylase, was catalyzed by the native FDC1. Expression of FDC1 was naturally induced by trans-cinnamate, the pathway intermediate and its substrate, at levels sufficient for ensuring flux through the pathway. Deletion of ARO10 to eliminate the competing Ehrlich pathway and expression of a feedback-resistant DAHP synthase encoded by ARO4(K229L) preserved and promoted the endogenous availability precursor phenylalanine, leading to improved pathway flux and styrene production. These systematic improvements allowed styrene titers to ultimately reach 29 mg/L at a glucose yield of 1.44 mg/g, a 60% improvement over the initial strain. CONCLUSIONS: The potential of S. cerevisiae as a host for renewable styrene production has been demonstrated. Significant strain improvements, however, will ultimately be needed to achieve economical production levels. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12934-014-0123-2) contains supplementary material, which is available to authorized users. BioMed Central 2014-08-21 /pmc/articles/PMC4145238/ /pubmed/25162943 http://dx.doi.org/10.1186/s12934-014-0123-2 Text en © McKenna et al.; licensee BioMed Central 2014 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research McKenna, Rebekah Thompson, Brian Pugh, Shawn Nielsen, David R Rational and combinatorial approaches to engineering styrene production by Saccharomyces cerevisiae |
title | Rational and combinatorial approaches to
engineering styrene production by Saccharomyces
cerevisiae |
title_full | Rational and combinatorial approaches to
engineering styrene production by Saccharomyces
cerevisiae |
title_fullStr | Rational and combinatorial approaches to
engineering styrene production by Saccharomyces
cerevisiae |
title_full_unstemmed | Rational and combinatorial approaches to
engineering styrene production by Saccharomyces
cerevisiae |
title_short | Rational and combinatorial approaches to
engineering styrene production by Saccharomyces
cerevisiae |
title_sort | rational and combinatorial approaches to
engineering styrene production by saccharomyces
cerevisiae |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4145238/ https://www.ncbi.nlm.nih.gov/pubmed/25162943 http://dx.doi.org/10.1186/s12934-014-0123-2 |
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