Todd French

Biodiesel production by in situ transesterification of municipal primary and secondary sludges

The potential of using municipal wastewater sludges as a lipid feedstock for biodiesel production was investigated. Primary and secondary sludge samples obtained from a municipal wastewater treatment plant in Tuscaloosa, AL were freeze-dried and subjected to an acid-catalyzed insitu transesterification process. Experiments were conducted to determine the effects of temperature, sulfuric acid concentration, and mass ratio of methanol to sludge on the yield of fatty acid methyl esters (FAMEs). Results indicated a significant interactive effect between temperature, acid concentration, and methanol to sludge mass ratio on the FAME yield for the insitu transesterification of primary sludge, while the FAME yield for secondary sludge was significantly affected by the independent effects of the three factors investigated. The maximum FAME yields were obtained at 75 degrees C, 5% (v/v) H(2)SO(4), and 12:1 methanol to sludge mass ratio and were 14.5% and 2.5% for primary and secondary sludge, respectively. Gas chromatography (GC) analysis of the FAMEs revealed a similar fatty acid composition for both primary and secondary sludge. An economic analysis estimated the cost of

Genome Sequence of the Oleaginous Yeast Rhodotorula glutinis ATCC 204091.

3.23/gallon for a neat biodiesel obtained from this process at an assumed yield of 10% FAMEs/dry weight of sludge.

Biocrude production by activated sludge microbial cultures using pulp and paper wastewaters as fermentation substrate

ABSTRACT Rhodotorula glutinis ATCC 204091 is an oleaginous oxidative red yeast that can accumulate lipids to >50% of its biomass when grown with appropriate carbon and nitrogen ratios. It produces a red pigment consisting of useful antioxidants, such as carotenoids, torulene, and torularhodin, when cultivated under carbon-deficient conditions.

Microbial Lipid Accumulation Capability of Activated Sludge Feeding on Short Chain Fatty Acids as Carbon Sources through Fed-Batch Cultivation

Municipal wastewater activated sludge contains a mixed microbial community, which can be manipulated to produce biocrude, a lipid feedstock for biodiesel production. In this study, the potential of biocrude production by activated sludge microorganisms grown in three different types of pulp and paper mill wastewaters was investigated. A 20% (v/v) activated sludge was inoculated into pulp and paper wastewater, supplemented with glucose (60 g/L) and nutrients (nitrogen and phosphorus) to obtain a high carbon to nitrogen ratio (70:1). The culture was incubated aerobically for seven days. The results showed that the activated sludge microorganisms were able to grow and accumulate lipids when cultivated in amended wastewaters. Microorganisms growing in anaerobic settling pond effluent water showed the highest lipid accumulation of up to 40.6% cell dry weight (CDW) after five days of cultivation compared with pulp wash wastewater (PuWW) (11.7% CDW) and mixed wastewater (MWW) (8.2% CDW) after seven days of cultivation. The lipids mostly contained C16‒C18 fatty acids groups with oleic acid and palmitic acid being the dominant fatty acids. The maximum biodiesel yield was about 6–8% CDW for all the wastewaters. The results showed the potential of utilizing pulp and paper mill effluents and other waste streams, such as activated sludge for the sustainable production of lipids for biofuel production.

Proteome and membrane fatty acid analyses on Oligotropha carboxidovorans OM5 grown under chemolithoautotrophic and heterotrophic conditions.

The potential of activated sludge microbial lipid technology as a sustainable energy platform has been recognized in the past years, but it has been challenged by the cost of carbon sources. This study hypothesized that Short Chain Fatty Acids (SCFAs) that can be derived from organic wastes can be alternative carbon sources. Therefore, this work evaluated the capability of activated sludge microbial consortia to accumulate microbial lipid by fed-batch feeding of SCFAs acetic acid, propionic acid, and butyric acid that were fed every 12 h period in 5 liter bioreactors. Activated sludge microbial consortia can accumulate microbial lipid by feeding on acetic acid. Acetic acid at 1.5 g/L loading per feeding enhanced the lipid content of activated sludge up to around 20% (w/w) dry biomass. This is comparable to that of oleaginous microorganisms. The feeding of nitrogen source (ammonium) at molar C/N of 70 only at the start resulted in significant lipid accumulation as compared to that from feeding of nitrogen for every feeding of the carbon source (acid-substrates). Fatty Acid Methyl Esters (FAMEs) profiles of the extracted lipids changed during cultivation. A biodiesel volumetric yield increase of 325% (w/w) from initial culture was achieved. This microbial lipid enhancement was confirmed using fluorescence microscopy imaging of neutral lipids, which also showed that the neutral lipid-containing cells are in the size range of yeasts. This work proved the hypothesis that activated sludge microbial consortia can accumulate microbial lipid by feeding on SCFAs.

Mississippi State Biodiesel Production Project

Oligotropha carboxidovorans OM5 T. (DSM 1227, ATCC 49405) is a chemolithoautotrophic bacterium able to utilize CO and H2 to derive energy for fixation of CO2. Thus, it is capable of growth using syngas, which is a mixture of varying amounts of CO and H2 generated by organic waste gasification. O. carboxidovorans is capable also of heterotrophic growth in standard bacteriologic media. Here we characterize how the O. carboxidovorans proteome adapts to different lifestyles of chemolithoautotrophy and heterotrophy. Fatty acid methyl ester (FAME) analysis of O. carboxidovorans grown with acetate or with syngas showed that the bacterium changes membrane fatty acid composition. Quantitative shotgun proteomic analysis of O. carboxidovorans grown in the presence of acetate and syngas showed production of proteins encoded on the megaplasmid for assimilating CO and H2 as well as proteins encoded on the chromosome that might have contributed to fatty acid and acetate metabolism. We found that adaptation to chemolithoautotrophic growth involved adaptations in cell envelope, oxidative homeostasis, and metabolic pathways such as glyoxylate shunt and amino acid/cofactor biosynthetic enzymes.

Supply Chain Network Model for Biodiesel Production via Wastewaters from Paper and Pulp Companies

Biodiesel is a renewable fuel conventionally generated from vegetable oils and animal fats that conforms to ASTM D6751. Depending on the free fatty acid content of the feedstock, biodiesel is produced via transesterification, esterification, or a combination of these processes. Currently the cost of the feedstock accounts for more than 80% of biodiesel production cost. The main goal of this project was to evaluate and develop non-conventional feedstocks and novel processes for producing biodiesel. One of the most novel and promising feedstocks evaluated involves the use of readily available microorganisms as a lipid source. Municipal wastewater treatment facilities (MWWTF) in the USA produce (dry basis) of microbial sludge annually. This sludge is composed of a variety of organisms, which consume organic matter in wastewater. The content of phospholipids in these cells have been estimated at 24% to 25% of dry mass. Since phospholipids can be transesterified they could serve as a ready source of biodiesel. Examination of the various transesterification methods shows that in situ conversion of lipids to FAMEs provides the highest overall yield of biodiesel. If one assumes a 7.0% overall yield of FAMEs from dry sewage sludge on a weight basis, the cost per gallon of extracted lipid would be

An Evaluation of the Feasibility of Combining Carbon Dioxide Flooding Technologies with Microbial Enhanced Oil Recovery Technologies in Order To Sequester Carbon Dioxide

3.11. Since the lipid is converted to FAMEs, also known as biodiesel, in the in Situ extraction process, the product can be used as is for renewable fuel. As transesterification efficiency increases the cost per gallon drops quickly, hitting

Extraction of lipids from municipal wastewater plant microorganisms for production of biodiesel

2.01 at 15.0% overall yield. An overall yield of 10.0% is required to obtain biodiesel at

Enhanced lipid and biodiesel production from glucose‐fed activated sludge: Kinetics and microbial community analysis

2.50 per gallon, allowing it to compete with soybean oil in the marketplace. Twelve plant species with potential for oil production were tested at Mississippi State, MS. Of the species tested, canola, rapeseed and birdseed rape appear to have potential in Mississippi as winter annual crops because of yield. Two perennial crops were investigated, Chinese tallow tree and tung tree. High seed yields from these species are possible because, there stature allows for a third dimension in yield (up). Harvest regimes have already been worked out with tung, and the large seed makes shedding of the seed with tree shakers possible. While tallow tree seed yields can be mind boggling (12,000 kg seed/ha at 40% oil), genotypes that shed seed easily are currently not known. Efficient methods were developed to isolate polyunsaturated fatty acid methyl esters from bio-diesel. The hypothesis to isolate this class of fatty acids, which are used as popular dietary supplements and prescription medicine (OMACOR), was that they bind transition metal ions much stronger than their harmful saturated analogs. AgBF4 has the highest extraction ability among all the metal ions tested. Glycerol is a key product from the production of biodiesel. It is produced during the transesterification process by cleaving the fatty acids from the glycerol backbone (the fatty acids are used as part of the biodiesel, which is a fatty acid methyl ester). Glycerol is a non-toxic compound with many uses; however, if a surplus exists in the future, more uses for the produced glycerol needs to be found. Another phase of the project was to find an add-on process to the biodiesel production process that will convert the glycerol by-product into more valuable substances for end uses other than food or cosmetics, focusing at present on 1,3-propanediol and lactic acid.All three MSU cultures produced products at concentrations below that of the benchmark microorganisms. There was one notable isolate the caught the eye of the investigators and that was culture J6 due to the ability of this microorganism to co-produce both products and one in particularly high concentrations. This culture with more understanding of its metabolic pathways could prove a useful biological agent for the conversion of glycerol. Heterogeneous catalysis was examined as an alternative to overcome the disadvantages of homogeneous transesterification, such as the presence of salts in the glycerine phase and the continuous lost of catalyst. A maximum soy biodiesel yield of 85% was obtained by BaO in 14 minutes, whereas, PbO, MnO2, CaO and MgO gave a maximum yields of 84%, 80%, 78% and 66% respectively at 215°C. The overall reaction order of PbO, MnO2, BaO, CaO and MgO was found to be 1, 1, 3, 1 and 1 respectively. The highest rate constant was observed for BaO, which was 0.0085 g2.mole-2.min-1. The performance of biodiesel in terms of type (e.g., NOx, and CO) and quantity of emissions was tested using soy biodiesel, blends of biodiesel and ethanol, and differently aged diesel engines. It was determined that saturated methyl esters, and relatively high oxygen content in the fuel, caused by addition of ethanol, increased the NOx emissions from new diesel engines compared to petroleum diesel.