Difference between revisions of "Team:TU Darmstadt/Demonstrate"
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==Proof of Concept== | ==Proof of Concept== | ||
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| + | ===Glycolic acid production in <i>E. coli </i>=== | ||
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| + | Our aim was to produce glycolic acid for the production of biodegradable polymers. We were able to overexpress the genes <i>aceA</i> and <i>ycdW</i> in Escherichia coli. The proteins AceA and YcdW were tagged with an His-tag and purified via an ÄKTA system. The purification was confirmed by an SDS-PAGE and western blot. After that we successfully characterized YcdW with an NADPH dependent enzyme assay. The assay indicates that YcdW uses glyoxylate as substrate under consumption of NADPH to produce glycolic acid. The functionality of AceA were verified with an phenylhydrazine-dependent assay. AceA catalyze the reaction from isocitrat to glyoxylate. We were able to detect glycolic acid via HPLC and therefore we produce one of our monomers. | ||
Revision as of 19:40, 12 October 2018
Proof of Concept
Glycolic acid production in E. coli
Our aim was to produce glycolic acid for the production of biodegradable polymers. We were able to overexpress the genes aceA and ycdW in Escherichia coli. The proteins AceA and YcdW were tagged with an His-tag and purified via an ÄKTA system. The purification was confirmed by an SDS-PAGE and western blot. After that we successfully characterized YcdW with an NADPH dependent enzyme assay. The assay indicates that YcdW uses glyoxylate as substrate under consumption of NADPH to produce glycolic acid. The functionality of AceA were verified with an phenylhydrazine-dependent assay. AceA catalyze the reaction from isocitrat to glyoxylate. We were able to detect glycolic acid via HPLC and therefore we produce one of our monomers.