Team:Wisconsin-Madison/biofuel

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Latest revision as of 22:58, 28 September 2011

Project >> Overview, Ethanol Sensor, Alkane Sensor, Microcompartment

Biofuels

Biofuel refers to any number of products that can be derived from biomass, and most commonly refers to alkyl esters (biodiesel) or alchohols, but can also refer to combustible gases and other hydrocarbons. The majority of biofuel research and production is focused on alternative sources of energy for transportation. Biofuel interest is primarily motivated from the need for more renewable fuels, due to the decreasing supply of petroleum and increasing gasoline prices. There is a lot of ongoing research into more efficient production of these kinds of biofuels.

Our goal this summer is to successfully engineer both ethanol and n-alkane biosensors to aid in the process of biofuel production. Biosensors could potentially offer the same sensing accuracy as current techniques, but for a lower cost. Our biosensors will use red fluorescent protein (RFP) to fluoresce in the presence of either ethanol or n-alkanes. The amount of RFP produced in the presence of either of these compounds will (ideally) be proportional to their concentration in solution.

Learn more about: genes

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                  <a href="https://2011.igem.org/Team:Wisconsin-Madison/directedevolution">Directed Evolution</a>
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 <strong><font size="3">
-Ethanol Sensor
+Biofuels
 </font></strong><p>
+Biofuel refers to any number of products that can be derived from biomass, and most commonly refers to alkyl esters (biodiesel) or alchohols, but can also refer to combustible gases and other hydrocarbons. The majority of <a href="http://www.glbrc.org/">biofuel research</a> and production is focused on alternative sources of energy for transportation. Biofuel interest is primarily motivated from the need for more renewable fuels, due to the decreasing supply of petroleum and increasing gasoline prices. There is a lot of ongoing research into more efficient production of these kinds of biofuels.
+<p>
+Our goal this summer is to successfully engineer both ethanol and n-alkane biosensors to aid in the process of biofuel production. Biosensors could potentially offer the same sensing accuracy as current techniques, but for a lower cost. Our biosensors will use red fluorescent protein (RFP) to fluoresce in the presence of either ethanol or n-alkanes. The amount of RFP produced in the presence of either of these compounds will (ideally) be proportional to their concentration in solution.
-The primary source of results thus far has been a plate reader. This is a device which can record the optical density (OD – a measure of cell concentration) as well as the fluorescence of up to 96 cultures at once. Ideally, we hope to produce such a robust fluorescent response as to not need the plate reader for qualitative assessments.
 <p><br>
-This graph shows the fluorescent response of cell cultures containing two plasmids: pBAD33bb: exaDE and PexaA: TagRFP in pSB1A2 (pEtRv2.1). The fluorescence of the system at varying ethanol concentrations was normalized to the optical density of their respective cell culture. The cells’ fluorescence was measured over a range of ethanol concentrations from 0%-5% EtOH. This ethanol sensing system is “turned on” or induced in the presence of arabinose by the PBAD promoter.  The blue data points on the graph represent the fluorescence of cell cultures without any arabinose and therefore the ethanol sensor is uninduced; the red data points represent the fluorescent response of the induced system in the presence of .2% arabinose.
+Learn more about: <a href="https://2011.igem.org/Team:Wisconsin-Madison/genes">genes</a>
-<p><br>
-<img src="https://static.igem.org/mediawiki/2011/0/00/Pets_2.1_graph.jpg" align="left" width="500">
-From the graph, it can be seen that there is an increase in fluorescence between the induced and uninduced system. This is the result that was expected, however, there isn’t a large difference in fluorescence between the two systems. To further increase this difference in fluorescence, we intend to decrease the “leakiness” of our promoter through directed evolution.  Additionally, the graph shows a positive correlation between fluorescence and ethanol concentration; therefore, our ethanol sensor exhibits a linear response in fluorescent intensity with varying ethanol concentrations. The magnitude of the slope of the data points represents the degree of our systems response. Currently, the slope of the data is smaller than desired. This will  also be the target of a directed evolution upon the ethanol sensor.