Team:Vanderbilt

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<p> <font size="3" face="georgia">Long before the advent of modern science, it was recognized that certain plants are capable of producing compounds of immense value. From a single class of molecule, the terpenoids, come properties including agents with therapeutic qualities against maladies ranging from cancer to infection, antimicrobials, natural pesticides, rich flavorants, and fragrant scents<sup>1</sup>. However, the utilization of these remarkable compounds has been severely hindered by their rarity in nature: many are found in only a small number of species and produced at levels measured in parts per million<sup>2</sup>. Synthetic biology offers an opportunity to resolve this problem, by applying metabolic engineering in order to create cellular factories. Our project seeks to use the ideas of synthetic biology to develop a commercially viable strategy for the efficient production of a wide range of terpenoids. </font> </p>
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<p>Long before the advent of modern science, it was recognized that certain plants are capable of producing compounds of immense value. From a single class of molecule, the terpenoids, come properties including agents with therapeutic qualities against maladies ranging from cancer to infection, antimicrobials, natural pesticides, rich flavorants, and fragrant scents<sup>1</sup>. However, the utilization of these remarkable compounds has been severely hindered by their rarity in nature: many are found in only a small number of species and produced at levels measured in parts per million<sup>2</sup>. Synthetic biology offers an opportunity to resolve this problem, by applying metabolic engineering in order to create cellular factories. Our project seeks to use the ideas of synthetic biology to develop a commercially viable strategy for the efficient production of a diverse range of terpenoids With our unique strategy, which begins in the greenhouse and ends in a bioreactor, we propose a way to efficiently and cost-effectively extract the genetic raw materials for terpene biosynthesis from a large number of species in parallel, thus allowing us to fulfill a previously neglected niche in the industry. </p>
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<p> <font size="3" face="georgia"> We believe that common brewer's yeast, or <i> Saccharomyces cerevisiae </i>, is an excellent platform for engineering terpenoid biosynthetic pathways. The mevalonic acid pathway endogenous to yeast produces the key isoprenoid intermediates that are the precursors to virtually all terpenoid biosynthesis<sup>3</sup>. Genes encoding plant synthases can then be recombinantly expressed in yeast cells, which then take that isoprenoid substrate and convert it through one or more steps into the final terpenoid product. A specially designed biobrick shuttle vector developed by our team should make the process convenient and reliable, by allowing us to first amplify plasmids containing our gene of interest in <i>E. coli</i> and permitting the integration of synthase genes directly into the yeast genome through homologous recombination. A carefully-refined protocol is expected to further improve product yield, by extracting genetic sequences directly from plant genomic DNA and mating cells to form diploid transformants. Combined, our approach promises to be an effective manufacturing platform for these precious (and sweet-smelling) compounds. </font>
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<p>We believe that common brewer's yeast, <i> Saccharomyces cerevisiae </i>, is an excellent platform for engineering terpenoid biosynthetic pathways. The mevalonic acid pathway endogenous to yeast produces the key isoprenoid intermediates that are the precursors to virtually all terpenoid biosynthesis<sup>3</sup>. Genes encoding plant synthases can then be recombinantly expressed in yeast cells, which then take that isoprenoid substrate and convert it through one or more steps into the final terpenoid product. We will take actual plants as our starting material, and extract the genes we need using protocols that are compatible with high-throughput processing of many species, including rare or endangered plants that may not have full genome sequence available. A specially designed biobrick shuttle vector developed by our team should make the process convenient and reliable, by allowing us to first amplify plasmids containing our gene of interest in <i>E. coli</i> and permitting the diploid integration of synthase genes directly into the yeast genome through homologous recombination. With the industry gravitating increasingly toward discovering unique combinations of terpenes, our contribution opens the way for efficiently expressing large quantities of different exotic terpenes. Combined, our approach promises to be an effective manufacturing platform for the production an almost limitless breadth of these precious (and sweet-smelling) compounds.
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<font size=4 face="georgia">See our <a href="http://2014.igem.org/Team:Vanderbilt/Project"style="color:#000000"> <u> project page</u></a> to learn more </font>
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<p>See our <a href="http://2014.igem.org/Team:Vanderbilt/Project"style="color:#000000"> <u> project page</u></a> to learn more </p>
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<b> References: </b><br>
<b> References: </b><br>
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1. Aharoni A, Jongsma MA, Bouwmeester HJ. Volatile science? Metabolic engineering of terpenoids in plants. Trends Plant Science 2005;10(12):594-602. </br>
1. Aharoni A, Jongsma MA, Bouwmeester HJ. Volatile science? Metabolic engineering of terpenoids in plants. Trends Plant Science 2005;10(12):594-602. </br>
2. Ajikumar PK, Tyo K, Carlsen S, Mucha O, Phon TH, Stephanopoulos G. Terpenoids: opportunities for biosynthesis of natural product drugs using engineered microorganisms. Molecular Pharmaceutics 2008;5(2):167-90. </br>
2. Ajikumar PK, Tyo K, Carlsen S, Mucha O, Phon TH, Stephanopoulos G. Terpenoids: opportunities for biosynthesis of natural product drugs using engineered microorganisms. Molecular Pharmaceutics 2008;5(2):167-90. </br>
3. Farhi M, Marhevka E, Masci T, Marcos E, Eyal Y, Ovadis M, Abeliovich H, Vainstein A. Harnessing yeast subcellular compartments for the production of plant terpenoids. Metabolic Engineering 2011;13(5):474-81.
3. Farhi M, Marhevka E, Masci T, Marcos E, Eyal Y, Ovadis M, Abeliovich H, Vainstein A. Harnessing yeast subcellular compartments for the production of plant terpenoids. Metabolic Engineering 2011;13(5):474-81.
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Latest revision as of 00:57, 9 February 2015

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Our Idea:

Our Approach:

Long before the advent of modern science, it was recognized that certain plants are capable of producing compounds of immense value. From a single class of molecule, the terpenoids, come properties including agents with therapeutic qualities against maladies ranging from cancer to infection, antimicrobials, natural pesticides, rich flavorants, and fragrant scents1. However, the utilization of these remarkable compounds has been severely hindered by their rarity in nature: many are found in only a small number of species and produced at levels measured in parts per million2. Synthetic biology offers an opportunity to resolve this problem, by applying metabolic engineering in order to create cellular factories. Our project seeks to use the ideas of synthetic biology to develop a commercially viable strategy for the efficient production of a diverse range of terpenoids With our unique strategy, which begins in the greenhouse and ends in a bioreactor, we propose a way to efficiently and cost-effectively extract the genetic raw materials for terpene biosynthesis from a large number of species in parallel, thus allowing us to fulfill a previously neglected niche in the industry.

We believe that common brewer's yeast, Saccharomyces cerevisiae , is an excellent platform for engineering terpenoid biosynthetic pathways. The mevalonic acid pathway endogenous to yeast produces the key isoprenoid intermediates that are the precursors to virtually all terpenoid biosynthesis3. Genes encoding plant synthases can then be recombinantly expressed in yeast cells, which then take that isoprenoid substrate and convert it through one or more steps into the final terpenoid product. We will take actual plants as our starting material, and extract the genes we need using protocols that are compatible with high-throughput processing of many species, including rare or endangered plants that may not have full genome sequence available. A specially designed biobrick shuttle vector developed by our team should make the process convenient and reliable, by allowing us to first amplify plasmids containing our gene of interest in E. coli and permitting the diploid integration of synthase genes directly into the yeast genome through homologous recombination. With the industry gravitating increasingly toward discovering unique combinations of terpenes, our contribution opens the way for efficiently expressing large quantities of different exotic terpenes. Combined, our approach promises to be an effective manufacturing platform for the production an almost limitless breadth of these precious (and sweet-smelling) compounds.


See our project page to learn more


References:
1. Aharoni A, Jongsma MA, Bouwmeester HJ. Volatile science? Metabolic engineering of terpenoids in plants. Trends Plant Science 2005;10(12):594-602.
2. Ajikumar PK, Tyo K, Carlsen S, Mucha O, Phon TH, Stephanopoulos G. Terpenoids: opportunities for biosynthesis of natural product drugs using engineered microorganisms. Molecular Pharmaceutics 2008;5(2):167-90.
3. Farhi M, Marhevka E, Masci T, Marcos E, Eyal Y, Ovadis M, Abeliovich H, Vainstein A. Harnessing yeast subcellular compartments for the production of plant terpenoids. Metabolic Engineering 2011;13(5):474-81.