Team:Johns Hopkins/Vit/Over

From 2011.igem.org

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(Overview)
(Overview)
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We demonstrated and characterized the production of pro Vitamin A, beta-carotene, in Saccharomyces cerevisiae, by importing three carotegenic genes from Xanthophyllomyces dendrorhous, namely carotene desaturase, GGPP synthase, phytoene synthase. The quantification was done via HPLC analysis and spectrophotometric assays over time. We also ran a comparative study of our yeast strains efficiency in dough versus in lab conditions using a novel substrate, dough media plates. We see production of significant amounts of betacarotene from this strain.
We demonstrated and characterized the production of pro Vitamin A, beta-carotene, in Saccharomyces cerevisiae, by importing three carotegenic genes from Xanthophyllomyces dendrorhous, namely carotene desaturase, GGPP synthase, phytoene synthase. The quantification was done via HPLC analysis and spectrophotometric assays over time. We also ran a comparative study of our yeast strains efficiency in dough versus in lab conditions using a novel substrate, dough media plates. We see production of significant amounts of betacarotene from this strain.
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[[File:VitA4.jpg|thumb|left|300px|GGPP synthase]]  
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[[File:VitA4.png|thumb|left|300px|GGPP synthase]] [[File:VitA2.png|thumb|left|300px|Betacarotene]]  
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[[File:VitA2.png|thumb|left|300px|Betacarotene]]  
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We have engineered a strain of saccharomyces cerevisiae to express L-ascorbic acid, or Vitamin C, for use in the baking of bread and brewing of beer. We codon optimized three genes from the Vitamin C pathway of arabidopsis thaliana and made biobrick compatible. These were then constructed denovo and assembled into expression cassettes. We are currently in the process of characterizing the Vitamin C production via HPLC and photospectrometric assays. We also in the process of constructing a combinatorial library of expression cassettes with 11 different strength promoters and terminators and have designed a selection and screening protocol to optimize production.   
We have engineered a strain of saccharomyces cerevisiae to express L-ascorbic acid, or Vitamin C, for use in the baking of bread and brewing of beer. We codon optimized three genes from the Vitamin C pathway of arabidopsis thaliana and made biobrick compatible. These were then constructed denovo and assembled into expression cassettes. We are currently in the process of characterizing the Vitamin C production via HPLC and photospectrometric assays. We also in the process of constructing a combinatorial library of expression cassettes with 11 different strength promoters and terminators and have designed a selection and screening protocol to optimize production.   

Revision as of 23:12, 24 September 2011

VitaYeast - Johns Hopkins University, iGEM 2011

Overview

We demonstrated and characterized the production of pro Vitamin A, beta-carotene, in Saccharomyces cerevisiae, by importing three carotegenic genes from Xanthophyllomyces dendrorhous, namely carotene desaturase, GGPP synthase, phytoene synthase. The quantification was done via HPLC analysis and spectrophotometric assays over time. We also ran a comparative study of our yeast strains efficiency in dough versus in lab conditions using a novel substrate, dough media plates. We see production of significant amounts of betacarotene from this strain.

GGPP synthase
Betacarotene

We have engineered a strain of saccharomyces cerevisiae to express L-ascorbic acid, or Vitamin C, for use in the baking of bread and brewing of beer. We codon optimized three genes from the Vitamin C pathway of arabidopsis thaliana and made biobrick compatible. These were then constructed denovo and assembled into expression cassettes. We are currently in the process of characterizing the Vitamin C production via HPLC and photospectrometric assays. We also in the process of constructing a combinatorial library of expression cassettes with 11 different strength promoters and terminators and have designed a selection and screening protocol to optimize production.