Theses and Dissertations

Issuing Body

Mississippi State University


Hernandez, Rafael

Committee Member

Magbanua, Benjamin

Committee Member

Elmore, Bill

Committee Member

Walters, Keisha

Committee Member

Bricka, R. Mark

Other Advisors or Committee Members

French, W. Todd

Date of Degree


Document Type

Dissertation - Open Access


Chemical Engineering

Degree Name

Doctor of Philosophy (Ph.D)


James Worth Bagley College of Engineering


Dave C. Swalm School of Chemical Engineering


The potential of enhancing lipid accumulation in municipal sewage activated sludge via fermentation of lignocellulose biomass hydrolyzate was investigated. The overall objective was to increase the levels of feedstock lipids in the activated sludge biomass and increase its biodiesel yield via in situ or ex situ transesterification; and improve its cost competitiveness as an abundant feedstock source for biofuels production. To reduce production costs and maintain environmental sustainability, influent wastewater and waste lignocellulose biomass hydrolyzate were used as cultivation media and substrate, respectively. However, lignocellulose hydrolyzates also contain degradation by-products such as furfural and acetic acid that are known to exert inhibitory effects on microorganisms; hence their effects on the fermentative performance of activated sludge were investigated and fermentation strategies were proposed and evaluated to counteract the microbial toxicity of these compounds. The utilization rate and efficiency of xylose by activated sludge microorganisms for lipid production was also evaluated as pentose sugars such as xylose usually constitute a major percentage of lignocellulose hydrolyzates. Furthermore, variations in the population profile of activated sludge microbiota were determined via 16S rRNA sequence analysis to determine the effect of sugar fermentation at different initial conditions. Results show that activated sludge lipid contents and biodiesel yield could be enhanced by fermentation of sugars at a high initial C:N ratio (70:1). Furfural was found to be highly inhibitory to microbial growth and lipid accumulation while high initial acetic acid concentrations enhanced biomass production but not lipid formation. Xylose was also utilized more efficiently than glucose by the activated sludge microorganisms for biomass and lipid production albeit at relatively slower rates; hence sugar mixtures derived from lignocellulose could be utilized for the process. Semicontinuous and continuous fermentation modes were proposed and evaluated as strategies to reduce inhibitory effect of furfural and acetic acid and improve lipid productivity, but the lack of nutrient supplementation prevented the cultures from sustaining microbial growth and lipid production, leading to cell death and washout. Finally, the reduction in the diversity of the activated sludge microbiota could point to specific microbial strains that are mainly responsible for lipid accumulation.