- Angela Zhu
- Steven Chang
- Samantha Alvarez
Title of Proposed Project
20.109(F12) Pre-Proposal: Engineering Alkyl Hydroperoxide Reductase Subunit C in E. coli to Extend the Oxidative Stability of Biodiesel Fuel
Biodiesel fuel is both a non-toxic and renewable resource, making it a promising alternative to standard diesel fuel. Current research focuses on synthesizing biodiesel from bacteria but does not address the poor oxidative stability of the biodiesel, which lowers its shelf life to a six month span. The proposed research would address this problem by engineering pre-existing biodiesel-producing bacteria to secrete antioxidative protein in order to prolong the oxidative stability of the biodiesel and therefore make it viable for a longer period of time.
Diesel fuel is a non-renewable resource made from a limited supply of fossil fuels, which is drawing attention towards more renewable biofuels like biodiesel . Biodiesel is primarily made of methyl esters, which are unsaturated fatty acid chains that are prone to oxidation via the free radical mechanism. It can be found in vegetable oils, animal fats, and even in used frying oil. The reason why biodiesel is currently not very commercially accepted, however, is because it is expensive and has poor oxidative stability when exposed to atmospheric oxygen when it is being stored . This degradation of fuel quality can affect properties like kinematic viscosity, acid value, and peroxide value . Treating the fatty derivatives with oxidation inhibitors, or antioxidants, prevents this premature degradation . Previous research in the field  has found that the addition of synthetic antioxidants into biodiesel improves oxidative stability. However, this process occurs synthetically through complex chemical processes in laboratories. Alternatively, by genetically engineering the biofuel-tolerant E. coli to these antioxidants, the bacteria eliminate the need for the overly complicated and expensive synthetic chemical process. This would result in a continuous supply of antioxidants for the biodiesel, which in the long run can be more cost-efficient because it prolongs the viability and stability of the biodiesel. With higher effiproduce ciency and lower cost in biodiesel production, commercial acceptance of biofuels will increase and therefore make it a more effective and environmentally friendly fuel source, which in the grand scheme of things can relieve the current over-dependence on nonrenewable fossil fuels.
 Dunn, Robert. O. "Effect of antioxidants on the oxidative stability of methyl soyate (biodiesel)." Fuel Processing Technology 86.10 (2005): 1071-1085.
 Knothe, Gerhard. "Some aspects of biodiesel oxidative stability." Fuel Processing Technology 88.7 (2007): 669-677.
 Meng, Xin and Jianming, et al. Yang. "Biodiesel production from oleaginous microorganism." Renewable Energy 34.1 (2009): 1-5.
 Tang, Haiying and Rhet C. et al. De Guzman. "The oxidative stability of biodiesel:Effects of FAME composition and antioxidant." Lipid Technology 20.11 (2008): 1-5.
Make clear what you see is the structural hole/gap in understanding or the need, and how you propose to fill in or satisfy what you've identified. You should specify your general approach (e.g. "will screen for mutants that enhance the contrast of the bacterial photography system") but do not need to think through the precise experimental details yet. Emphasize instead what results hope to collect and how they might improve the shortcomings that you've identified as interesting.
The two components to this research design are genetically engineering E. coli to secrete Alkyl Hydroperoxide Reductase Subunit C and testing the ability of the secreted Alkyl Hydroperoxide Reductase Subunit C to increase oxidative stability in biodiesel samples at varying concentrations. The Alkyl Hydroperoxide Reductase Subunit C genetic sequence from ______ E. coli (XXXXX) will be inserted into a plasmid with specific restriction enzyme markers? and transformed into biodiesel-producing? E. coli of a different strain?. The ability of the E. coli to express the gene function and secrete the Alkyl Hydroperoxide Reductase Subunit C will be tested and tracked with GFP. After successfully secreting the Alkyl Hydroperoxide Reductase Subunit C, its effectiveness to increase the stability of the biodiesel (as compared to a control)will be tested as in previous research. Future work would be to make the E. coli bacteria more tolerable to the conditions needed to survive in a biodiesel fuel tank because bla bla bla.
Schematic of AhpC Production
Dunn, Robert. O. "Effect of antioxidants on the oxidative stability of methyl soyate (biodiesel)." Fuel Processing Technology 86.10 (2005): 1071-1085.
Knothe, Gerhard. "Some aspects of biodiesel oxidative stability." Fuel Processing Technology 88.7 (2007): 669-677.
Meng, Xin and Jianming, et al. Yang. "Biodiesel production from oleaginous microorganism." Renewable Energy 34.1 (2009): 1-5.
Tang, Haiying and Rhet C. et al. De Guzman. "The oxidative stability of biodiesel:Effects of FAME composition and antioxidant." Lipid Technology 20.11 (2008): 1-5.