Biosynthesis of Vitamin C by Yeast Leads to Increased Stress Resistance
In industrial large scale bio-reactions micro-organisms are generally exposed to a variety of environmental stresses, which might be detrimental for growth and productivity. Reactive oxygen species (ROS) play a key role among the common stress factors--directly--through incomplete reduction of O(2)...
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| Published in | PloS one Vol. 2; no. 10; p. e1092 |
|---|---|
| Main Authors | , , , , , |
| Format | Journal Article |
| Language | English |
| Published |
United States
Public Library of Science
31.10.2007
Public Library of Science (PLoS) |
| Subjects | |
| Online Access | Get full text |
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| Abstract | In industrial large scale bio-reactions micro-organisms are generally exposed to a variety of environmental stresses, which might be detrimental for growth and productivity. Reactive oxygen species (ROS) play a key role among the common stress factors--directly--through incomplete reduction of O(2) during respiration, or indirectly--caused by other stressing factors. Vitamin C or L-ascorbic acid acts as a scavenger of ROS, thereby potentially protecting cells from harmful oxidative products. While most eukaryotes synthesize ascorbic acid, yeast cells produce erythro-ascorbic acid instead. The actual importance of this antioxidant substance for the yeast is still a subject of scientific debate.
We set out to enable Saccharomyces cerevisiae cells to produce ascorbic acid intracellularly to protect the cells from detrimental effects of environmental stresses. We report for the first time the biosynthesis of L-ascorbic acid from D-glucose by metabolically engineered yeast cells. The amount of L-ascorbic acid produced leads to an improved robustness of the recombinant cells when they are subjected to stress conditions as often met during industrial fermentations. Not only resistance against oxidative agents as H(2)O(2) is increased, but also the tolerance to low pH and weak organic acids at low pH is increased.
This platform provides a new tool whose commercial applications may have a substantial impact on bio-industrial production of Vitamin C. Furthermore, we propose S. cerevisiae cells endogenously producing vitamin C as a cellular model to study the genesis/protection of ROS as well as genotoxicity. |
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| AbstractList | Background
In industrial large scale bio-reactions micro-organisms are generally exposed to a variety of environmental stresses, which might be detrimental for growth and productivity. Reactive oxygen species (ROS) play a key role among the common stress factors–directly-through incomplete reduction of O2 during respiration, or indirectly-caused by other stressing factors. Vitamin C or L-ascorbic acid acts as a scavenger of ROS, thereby potentially protecting cells from harmful oxidative products. While most eukaryotes synthesize ascorbic acid, yeast cells produce erythro-ascorbic acid instead. The actual importance of this antioxidant substance for the yeast is still a subject of scientific debate.
Methodology/Principal Findings
We set out to enable Saccharomyces cerevisiae cells to produce ascorbic acid intracellularly to protect the cells from detrimental effects of environmental stresses. We report for the first time the biosynthesis of L-ascorbic acid from D-glucose by metabolically engineered yeast cells. The amount of L-ascorbic acid produced leads to an improved robustness of the recombinant cells when they are subjected to stress conditions as often met during industrial fermentations. Not only resistance against oxidative agents as H2O2 is increased, but also the tolerance to low pH and weak organic acids at low pH is increased.
Conclusions/Significance
This platform provides a new tool whose commercial applications may have a substantial impact on bio-industrial production of Vitamin C. Furthermore, we propose S. cerevisiae cells endogenously producing vitamin C as a cellular model to study the genesis/protection of ROS as well as genotoxicity. Background In industrial large scale bio-reactions micro-organisms are generally exposed to a variety of environmental stresses, which might be detrimental for growth and productivity. Reactive oxygen species (ROS) play a key role among the common stress factors-directly-through incomplete reduction of O.sub.2 during respiration, or indirectly-caused by other stressing factors. Vitamin C or L-ascorbic acid acts as a scavenger of ROS, thereby potentially protecting cells from harmful oxidative products. While most eukaryotes synthesize ascorbic acid, yeast cells produce erythro-ascorbic acid instead. The actual importance of this antioxidant substance for the yeast is still a subject of scientific debate. Methodology/Principal Findings We set out to enable Saccharomyces cerevisiae cells to produce ascorbic acid intracellularly to protect the cells from detrimental effects of environmental stresses. We report for the first time the biosynthesis of L-ascorbic acid from D-glucose by metabolically engineered yeast cells. The amount of L-ascorbic acid produced leads to an improved robustness of the recombinant cells when they are subjected to stress conditions as often met during industrial fermentations. Not only resistance against oxidative agents as H.sub.2 O.sub.2 is increased, but also the tolerance to low pH and weak organic acids at low pH is increased. Conclusions/Significance This platform provides a new tool whose commercial applications may have a substantial impact on bio-industrial production of Vitamin C. Furthermore, we propose S. cerevisiae cells endogenously producing vitamin C as a cellular model to study the genesis/protection of ROS as well as genotoxicity. In industrial large scale bio-reactions micro-organisms are generally exposed to a variety of environmental stresses, which might be detrimental for growth and productivity. Reactive oxygen species (ROS) play a key role among the common stress factors--directly--through incomplete reduction of O(2) during respiration, or indirectly--caused by other stressing factors. Vitamin C or L-ascorbic acid acts as a scavenger of ROS, thereby potentially protecting cells from harmful oxidative products. While most eukaryotes synthesize ascorbic acid, yeast cells produce erythro-ascorbic acid instead. The actual importance of this antioxidant substance for the yeast is still a subject of scientific debate. We set out to enable Saccharomyces cerevisiae cells to produce ascorbic acid intracellularly to protect the cells from detrimental effects of environmental stresses. We report for the first time the biosynthesis of L-ascorbic acid from D-glucose by metabolically engineered yeast cells. The amount of L-ascorbic acid produced leads to an improved robustness of the recombinant cells when they are subjected to stress conditions as often met during industrial fermentations. Not only resistance against oxidative agents as H(2)O(2) is increased, but also the tolerance to low pH and weak organic acids at low pH is increased. This platform provides a new tool whose commercial applications may have a substantial impact on bio-industrial production of Vitamin C. Furthermore, we propose S. cerevisiae cells endogenously producing vitamin C as a cellular model to study the genesis/protection of ROS as well as genotoxicity. In industrial large scale bio-reactions micro-organisms are generally exposed to a variety of environmental stresses, which might be detrimental for growth and productivity. Reactive oxygen species (ROS) play a key role among the common stress factors--directly--through incomplete reduction of O(2) during respiration, or indirectly--caused by other stressing factors. Vitamin C or L-ascorbic acid acts as a scavenger of ROS, thereby potentially protecting cells from harmful oxidative products. While most eukaryotes synthesize ascorbic acid, yeast cells produce erythro-ascorbic acid instead. The actual importance of this antioxidant substance for the yeast is still a subject of scientific debate.BACKGROUNDIn industrial large scale bio-reactions micro-organisms are generally exposed to a variety of environmental stresses, which might be detrimental for growth and productivity. Reactive oxygen species (ROS) play a key role among the common stress factors--directly--through incomplete reduction of O(2) during respiration, or indirectly--caused by other stressing factors. Vitamin C or L-ascorbic acid acts as a scavenger of ROS, thereby potentially protecting cells from harmful oxidative products. While most eukaryotes synthesize ascorbic acid, yeast cells produce erythro-ascorbic acid instead. The actual importance of this antioxidant substance for the yeast is still a subject of scientific debate.We set out to enable Saccharomyces cerevisiae cells to produce ascorbic acid intracellularly to protect the cells from detrimental effects of environmental stresses. We report for the first time the biosynthesis of L-ascorbic acid from D-glucose by metabolically engineered yeast cells. The amount of L-ascorbic acid produced leads to an improved robustness of the recombinant cells when they are subjected to stress conditions as often met during industrial fermentations. Not only resistance against oxidative agents as H(2)O(2) is increased, but also the tolerance to low pH and weak organic acids at low pH is increased.METHODOLOGY/PRINCIPAL FINDINGSWe set out to enable Saccharomyces cerevisiae cells to produce ascorbic acid intracellularly to protect the cells from detrimental effects of environmental stresses. We report for the first time the biosynthesis of L-ascorbic acid from D-glucose by metabolically engineered yeast cells. The amount of L-ascorbic acid produced leads to an improved robustness of the recombinant cells when they are subjected to stress conditions as often met during industrial fermentations. Not only resistance against oxidative agents as H(2)O(2) is increased, but also the tolerance to low pH and weak organic acids at low pH is increased.This platform provides a new tool whose commercial applications may have a substantial impact on bio-industrial production of Vitamin C. Furthermore, we propose S. cerevisiae cells endogenously producing vitamin C as a cellular model to study the genesis/protection of ROS as well as genotoxicity.CONCLUSIONS/SIGNIFICANCEThis platform provides a new tool whose commercial applications may have a substantial impact on bio-industrial production of Vitamin C. Furthermore, we propose S. cerevisiae cells endogenously producing vitamin C as a cellular model to study the genesis/protection of ROS as well as genotoxicity. In industrial large scale bio-reactions micro-organisms are generally exposed to a variety of environmental stresses, which might be detrimental for growth and productivity. Reactive oxygen species (ROS) play a key role among the common stress factors-directly-through incomplete reduction of O.sub.2 during respiration, or indirectly-caused by other stressing factors. Vitamin C or L-ascorbic acid acts as a scavenger of ROS, thereby potentially protecting cells from harmful oxidative products. While most eukaryotes synthesize ascorbic acid, yeast cells produce erythro-ascorbic acid instead. The actual importance of this antioxidant substance for the yeast is still a subject of scientific debate. We set out to enable Saccharomyces cerevisiae cells to produce ascorbic acid intracellularly to protect the cells from detrimental effects of environmental stresses. We report for the first time the biosynthesis of L-ascorbic acid from D-glucose by metabolically engineered yeast cells. The amount of L-ascorbic acid produced leads to an improved robustness of the recombinant cells when they are subjected to stress conditions as often met during industrial fermentations. Not only resistance against oxidative agents as H.sub.2 O.sub.2 is increased, but also the tolerance to low pH and weak organic acids at low pH is increased. This platform provides a new tool whose commercial applications may have a substantial impact on bio-industrial production of Vitamin C. Furthermore, we propose S. cerevisiae cells endogenously producing vitamin C as a cellular model to study the genesis/protection of ROS as well as genotoxicity. Background In industrial large scale bio-reactions micro-organisms are generally exposed to a variety of environmental stresses, which might be detrimental for growth and productivity. Reactive oxygen species (ROS) play a key role among the common stress factors–directly-through incomplete reduction of O2 during respiration, or indirectly-caused by other stressing factors. Vitamin C or L-ascorbic acid acts as a scavenger of ROS, thereby potentially protecting cells from harmful oxidative products. While most eukaryotes synthesize ascorbic acid, yeast cells produce erythro-ascorbic acid instead. The actual importance of this antioxidant substance for the yeast is still a subject of scientific debate. Methodology/Principal Findings We set out to enable Saccharomyces cerevisiae cells to produce ascorbic acid intracellularly to protect the cells from detrimental effects of environmental stresses. We report for the first time the biosynthesis of L-ascorbic acid from D-glucose by metabolically engineered yeast cells. The amount of L-ascorbic acid produced leads to an improved robustness of the recombinant cells when they are subjected to stress conditions as often met during industrial fermentations. Not only resistance against oxidative agents as H2O2 is increased, but also the tolerance to low pH and weak organic acids at low pH is increased. Conclusions/Significance This platform provides a new tool whose commercial applications may have a substantial impact on bio-industrial production of Vitamin C. Furthermore, we propose S. cerevisiae cells endogenously producing vitamin C as a cellular model to study the genesis/protection of ROS as well as genotoxicity. BACKGROUND: In industrial large scale bio-reactions micro-organisms are generally exposed to a variety of environmental stresses, which might be detrimental for growth and productivity. Reactive oxygen species (ROS) play a key role among the common stress factors--directly--through incomplete reduction of O(2) during respiration, or indirectly--caused by other stressing factors. Vitamin C or L-ascorbic acid acts as a scavenger of ROS, thereby potentially protecting cells from harmful oxidative products. While most eukaryotes synthesize ascorbic acid, yeast cells produce erythro-ascorbic acid instead. The actual importance of this antioxidant substance for the yeast is still a subject of scientific debate. METHODOLOGY/PRINCIPAL FINDINGS: We set out to enable Saccharomyces cerevisiae cells to produce ascorbic acid intracellularly to protect the cells from detrimental effects of environmental stresses. We report for the first time the biosynthesis of L-ascorbic acid from D-glucose by metabolically engineered yeast cells. The amount of L-ascorbic acid produced leads to an improved robustness of the recombinant cells when they are subjected to stress conditions as often met during industrial fermentations. Not only resistance against oxidative agents as H(2)O(2) is increased, but also the tolerance to low pH and weak organic acids at low pH is increased. CONCLUSIONS/SIGNIFICANCE: This platform provides a new tool whose commercial applications may have a substantial impact on bio-industrial production of Vitamin C. Furthermore, we propose S. cerevisiae cells endogenously producing vitamin C as a cellular model to study the genesis/protection of ROS as well as genotoxicity. |
| Audience | Academic |
| Author | Sauer, Michael Fossati, Tiziana Mattanovich, Diethard Branduardi, Paola Pagani, Roberto Porro, Danilo |
| AuthorAffiliation | Baylor College of Medicine, United States of America 1 Dipartimento di Biotecnologie e Bioscienze, Università degli Studi di Milano-Bicocca, Milano, Italy 3 Institute of Applied Microbiology, University of Natural Resources and Applied Life Sciences, Vienna, Austria 2 School of Bioengineering, University of Applied Sciences, Vienna, Austria |
| AuthorAffiliation_xml | – name: Baylor College of Medicine, United States of America – name: 2 School of Bioengineering, University of Applied Sciences, Vienna, Austria – name: 3 Institute of Applied Microbiology, University of Natural Resources and Applied Life Sciences, Vienna, Austria – name: 1 Dipartimento di Biotecnologie e Bioscienze, Università degli Studi di Milano-Bicocca, Milano, Italy |
| Author_xml | – sequence: 1 givenname: Paola surname: Branduardi fullname: Branduardi, Paola – sequence: 2 givenname: Tiziana surname: Fossati fullname: Fossati, Tiziana – sequence: 3 givenname: Michael surname: Sauer fullname: Sauer, Michael – sequence: 4 givenname: Roberto surname: Pagani fullname: Pagani, Roberto – sequence: 5 givenname: Diethard surname: Mattanovich fullname: Mattanovich, Diethard – sequence: 6 givenname: Danilo surname: Porro fullname: Porro, Danilo |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/17971855$$D View this record in MEDLINE/PubMed |
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| ContentType | Journal Article |
| Copyright | COPYRIGHT 2007 Public Library of Science 2007 Branduardi et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License. Branduardi et al. 2007 |
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| DocumentTitleAlternate | Cloning the Vitamin C Pathway |
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| Notes | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 14 content type line 23 Conceived and designed the experiments: DP PB MS DM. Performed the experiments: PB TF RP. Analyzed the data: DP PB TF RP MS DM. Wrote the paper: DP PB TF MS DM. |
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| Snippet | In industrial large scale bio-reactions micro-organisms are generally exposed to a variety of environmental stresses, which might be detrimental for growth and... Background In industrial large scale bio-reactions micro-organisms are generally exposed to a variety of environmental stresses, which might be detrimental for... BACKGROUND: In industrial large scale bio-reactions micro-organisms are generally exposed to a variety of environmental stresses, which might be detrimental... Background In industrial large scale bio-reactions micro-organisms are generally exposed to a variety of environmental stresses, which might be detrimental for... |
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| SubjectTerms | Animals Antioxidants Arabidopsis thaliana Ascorbic acid Ascorbic Acid - biosynthesis Ascorbic Acid - chemistry Baking yeast Biochemistry/Biocatalysis Bioengineering Biosynthesis Biotechnology Biotechnology/Applied Microbiology Biotechnology/Biocatalysis Biotechnology/Bioengineering Biotechnology/Plant Biotechnology Cell Biology/Cell Growth and Division Cell Biology/Cellular Death and Stress Responses Cell Biology/Microbial Growth and Development Cell Biology/Microbial Physiology and Metabolism Cloning Dehydrogenases Deoxyribonucleic acid Diastereomers DNA Drug resistance E coli Environmental effects Environmental stress Enzymes Escherichia coli Eukaryotes Fermentation Fungal Proteins - chemistry Genotoxicity Glucose Glucose - metabolism Hydrogen peroxide Hydrogen Peroxide - chemistry Hydrogen-Ion Concentration Industrial production Laboratories Life sciences Lung cancer Metabolism Microbiology Microbiology/Applied Microbiology Microbiology/Microbial Growth and Development Microbiology/Microbial Physiology and Metabolism Microscopy, Fluorescence Models, Biological Natural resources Organic acids Organisms Oxidation resistance Oxidative stress Oxygen Oxygen - metabolism pH effects Phosphatase Physiological aspects Plasmids Polyethylene glycol Rats Reactive Oxygen Species Recombinant Proteins - chemistry Saccharomyces cerevisiae Saccharomyces cerevisiae - metabolism Stresses Vitamin C Yeast |
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| Title | Biosynthesis of Vitamin C by Yeast Leads to Increased Stress Resistance |
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