Compartmental separation of Chlamydomonas reinhardtii for MFA
Abstract
A depleting source of fossil fuels is causing the search for new and renewable fuels to power our vehicles. Biodiesel, a renewable fuel, can be produced from the oil extracts of plants and algae. When compared to oil crops, oil from algae consists of a larger fraction of the organism’s biomass and requires less time for its production. Land required for oil production via oil crops would also compete with land used for food; indicating algae’s potential as a viable oil source. The main factor inhibiting algae’s use is the economically unfavorable process required for its growth and harvesting. Cost can be reduced through using genetic engineering to change the metabolism of the algal cell to increase the amount of oil it can store. This engineering requires a detailed knowledge of the metabolic rates of the cell’s metabolism. Experimentally determining these rates requires the separation of the cellular compartments of the algae. For this study, compartmental separation of the algal strain Chlamydomonas reinhardtii was examined through the use of non-aqueous fractionation. This examination also included the method required for rupturing the cell as well as the formation of the non-aqueous gradient needed for the separation of the cellular compartments. This study also analyzed the method necessary for determining compartmental separations. The work of this study lead to the determination of the methods required for the rupturing of the cells, non-aqueous fractionation of the cellular material, and western blotting of the protein extracts of the fractionated material. The separation of the cellular material through non-aqueous fractionation was indicated through the use of these western blots. A smaller study was performed on the effects of nitrogen starvation on the spectrophotometric characteristics of the algal cells when in mixotrophic and autotrophic conditions. It was found that nitrogen starvation in mixotrophic conditions changed the algae’s cell dry weight concentration to absorbance ratio while autotrophic growth showed no effect.
Degree
M.S.Ch.E.
Advisors
Morgan, Purdue University.
Subject Area
Chemical engineering
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