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Team:Cambridge/Bioluminescence/Luciferin Regeneration
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| - | Fireflies are easily visible in the evening, whereas colonies of bacteria expressing luciferase are not. We hope to remove this divide by expressing further enzymes exploited by fireflies. In <i>E. coli</i> and in eukaryotes, each molecule of luciferin that emits light is turned into oxyluciferin and cannot be regenerated. A further problem is that this oxyluciferin inhibits the further reactions of luciferase | + | ==Introduction== |
| + | Fireflies are easily visible in the evening, whereas colonies of bacteria expressing luciferase are not. We hope to remove this divide by expressing further enzymes exploited by fireflies. In <i>E. coli</i> and in eukaryotes, each molecule of luciferin that emits light is turned into oxyluciferin and cannot be regenerated. A further problem is that this oxyluciferin inhibits the further reactions of luciferase. | ||
| - | + | [http://www.ncbi.nlm.nih.gov/pubmed/11457857 Gomi and Kajiyama (2001)] describe a <strong>luciferin regenerating enzyme (LRE)</strong>, which we believed may solve these problems and allow brighter longer lasting light output without further addition of substrate. | |
[[Image:Cam-luci-cycle.jpg|center|500px]] | [[Image:Cam-luci-cycle.jpg|center|500px]] | ||
| + | LRE transforms the inhibitory oxyluciferin to CHBT, which is converted back into the luciferin substrate required for light output. This was believed to occur in one of two ways: non-enzymatically requiring D-cysteine, or via L-luciferin requiring the L-cysteine '''ASK PAUL'''. We aimed to verify these claims in our lab experiments and to quantify the difference in light output that LRE makes when expressed in ''E. coli''. | ||
| + | |||
| + | ==Our Experiments== | ||
{{:Team:Cambridge/Templates/footer}} | {{:Team:Cambridge/Templates/footer}} | ||
Revision as of 15:17, 23 October 2010
Artist's impression of what we would like
to achieve
Introduction
Fireflies are easily visible in the evening, whereas colonies of bacteria expressing luciferase are not. We hope to remove this divide by expressing further enzymes exploited by fireflies. In E. coli and in eukaryotes, each molecule of luciferin that emits light is turned into oxyluciferin and cannot be regenerated. A further problem is that this oxyluciferin inhibits the further reactions of luciferase.
Gomi and Kajiyama (2001) describe a luciferin regenerating enzyme (LRE), which we believed may solve these problems and allow brighter longer lasting light output without further addition of substrate.
LRE transforms the inhibitory oxyluciferin to CHBT, which is converted back into the luciferin substrate required for light output. This was believed to occur in one of two ways: non-enzymatically requiring D-cysteine, or via L-luciferin requiring the L-cysteine ASK PAUL. We aimed to verify these claims in our lab experiments and to quantify the difference in light output that LRE makes when expressed in E. coli.
Our Experiments
