Date of Graduation

5-2011

Document Type

Thesis

Degree Name

Bachelor of Science in Biological Engineering

Degree Level

Undergraduate

Department

Biological and Agricultural Engineering

Advisor

Hestekin, Jamie A

Abstract

As the demand for alternative fuels steadily increases, algae continue to be an excellent source for the development of biofuels. While algae have exhibited substantial potential for butanol production, there are limitations when it comes to the extraction process-its inability to withdraw oils and sugars usable for fermentation from cell walls. The genes of two bacteria were combined, creating a new organism that can both extract sugars and oils from algal cell walls and create butanol, simplifying the fermentation process while increasing efficiency. Pseudomonas flourescens, an obligate aerobe, has been shown in literature to degrade these cell walls to make sugars and oils available for fermentation. An anaerobic bacterium, Clostridium acetobutylicum, is commonly used for biofuel production because its ability to make biobutanol, among other biofuels. P. flourescens is absent one gene, alcohol dehydrogenase (AdhE2), from C. acetobutylicum's biobutanol pathway. The AdhE2 gene was extracted from C. acetobutylicum using PCR and then cloned it into P. flourescens using the pCN51 shuttle vector. The newly created organism that can both create biobutanol and perform algae lysis was cultured in glucose rich nutrient broth. Biobutanol production was confirmed using gas chromatography and HPLC

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