Team Project Description

With the increase in carbon dioxide emissions and fuel prices, alternatives to fossil fuels have become increasingly important. As a renewable resource with a wide range of energy densities, biofuels are a promising alternative. Synthetic biology has made it possible to manipulate select bacteria that naturally produce such fuels in order to improve the quantity and expand the range of hydrocarbons produced to achieve a higher production yield. The chassis selected for this project is Clostridium acetobutylicum, an anaerobe and native alcohol producer. The focus of our research currently consists of developing a shuttle vector for use in both Escherichia coli and C. acetobutylicum.

Continuing from previous year’s work, the shuttle vector has been assembled with the following parts: clostridial origin of replication (clos ori), a ptb (phosphotransbutyrylase) promoter from C. acetobutylicum, macrolide-lincosamide-streptogramin B resistance (mlsR) selection marker, a terminator from the iGEM registry, and the iGEM pSB1C3 backbone.

Taking advantage of the native acetone, butanol, and ethanol (ABE) fermentation pathways present in C. acetobutylicum, we plan to use our shuttle vector to introduce a combination of genes to produce branched and longer chain hydrocarbons. By operating these synthetic pathways in our anaerobic chassis and comparing them to production in non-native alcohol producing aerobic organisms, such as E. coli, we hope to address the overall question of whether or not anaerobes ‘do it better’.