W. Todd French

The effect of glycerol as a sole and secondary substrate on the growth and fatty acid composition of Rhodotorula glutinis

Rhodotorula glutinis is a yeast that produces copious quantities of lipids in the form of triacylglycerols (TAG) and can be used to make biodiesel via a transesterification process. The ester bonds in the TAG are broken leaving behind two products: fatty acid methyl esters and glycerol that could provide an inexpensive carbon source to grow oleaginous yeast R. glutinis. Described here are the effects of different growth substrates on TAG accumulation and fatty acids produced by R. glutinis. Yeast cultured 24h on medium containing dextrose, xylose, glycerol, dextrose and xylose, xylose and glycerol, or dextrose and glycerol accumulated 16, 12, 25, 10, 21, and 34% TAG on a dry weight basis, respectively. Lipids were extracted from R. glutinis culture and transesterified to form fatty acid methyl esters. The results show a difference in the degree of saturation for the carbon sources tested. Cells cultivated on glycerol alone had the highest degree of unsaturated fatty acids at 53% while xylose had the lowest at 25%. R. glutinis can be cultivated on all sugars tested as single carbon substrates or in mixtures. Glycerol may be used as secondary or primary carbon substrate.

Methanogenesis under acidic pH conditions in a semi-continuous reactor system

Operating an anaerobic digester at low pH could offer several advantages over operation at neutral pH. Most wastewater streams targeted for anaerobic digestion are inherently acidic, requiring alkalinity supplementation (at added expense) to buffer the pH at neutral. Additionally, previously published work completed by the authors using batch systems suggested that lowering the system pH could increase methane production by as much as 30%. The goal of this research was to evaluate the feasibility of sustaining methanogenesis at low pH in a semi-continuous laboratory-scale fermentor. Significant methane production was achieved in a system ranging in pH from approximately 4.0-5.3. Results show that, if the consortium is allowed to sufficiently acclimate to acidic conditions, methanogenesis can be maintained under acidic pH conditions, resulting in overall chemical oxygen demand (COD) reduction and methane production comparable to that achieved in a neutral pH system.

Lipid-enhancement of activated sludges obtained from conventional activated sludge and oxidation ditch processes

Lipid-enhancement of activated sludges was conducted to increase the amount of saponifiable lipids in the sludges. The sludges were obtained from a conventional activated sludge (CAS) and an oxidation ditch process (ODP). Results showed 59-222% and 150-250% increase in saponifiable lipid content of the sludges from CAS and ODP, respectively. The fatty acid methyl ester (FAMEs) obtained from triacylglycerides was 57-67% (of total FAMEs) for enhanced CAS and 55-73% for enhanced ODP, a very significant improvement from 6% to 10% (CAS) and 4% to 8% (ODP). Regardless of the source, the enhancement resulted in sludges with similar fatty acid profile indicating homogenization of the lipids in the sludges. This study provides a potential strategy to utilize existing wastewater treatment facilities as source of significant amount of lipids for biofuel applications.

Parametric Study on the Production of Renewable Fuels and Chemicals from Phospholipid-Containing Biomass

Heterogeneous catalytic cracking of lipids into transportation fuels and other specialty chemicals offers a unique opportunity to provide sustainable energy while utilizing the current petroleum infrastructure. This study addresses a possible route for the utilization of phospholipid-containing biomass for production of renewable fuels and chemicals. The first part of the study focused on reaction pathways associated with the catalytic cracking of model phospholipid into fuel-type compounds. The results indicated that phospholipid cracking proceeds via acid-catalyzed mechanisms, which resulted to cleavage of fatty acids and glycerol moieties. Microbial lipids, which contain large concentrations of phospholipids, are a potential non-food related feedstock to displace petroleum. Among possible sources of microbial lipids, activated sludge offers a distinct advantage for its availability and abundance at a potential fee. The second part of this work dealt with the conversion of activated sludge to chemicals of value as fuel components through fluidized-bed catalytic cracking. The effect of moisture level and catalyst loading were determined. Results indicated that moisture level of up to 15 % (weight) has no effect on total product yield. On the other hand, higher catalyst loading resulted to a higher yield of gaseous product. Significant coke deposition was inferred as indicated by the high proportion of phenolic compounds produced from pyrolytic reactions. The results of this study provide a potential route of utilizing bulk lipid feedstocks, without the removal of phosphorus-containing molecules, to produce fuel components.

Isopropanol and acetone induces vinyl chloride degradation in Rhodococcus rhodochrous

Abstract In situ bioremediation of vinyl chloride (VC)-contaminated waste sites requires a microorganism capable of degrading VC. While propane will induce an oxygenase to accomplish this goal, its use as a primary substrate in bioremediation is complicated by its flammability and low water solubility. This study demonstrates that two degradation products of propane, isoproponal and acetone, can induce the enzymes in Rhodococcus rhodochrous that degrade VC. Additionally, a reasonable number of cells for bioremediation can be grown on conventional solid bacteriological media (nutrient agar, tryptic soy agar, plate count agar) in an average microbiological laboratory and then induced to produce the necessary enzymes by incubation of a resting cell suspension with isopropanol or acetone. Since acetone is more volatile than isopropanol and has other undesirable characteristics, isopropanol is the inducer of choice. It offers a non-toxic, water-soluble, relatively inexpensive alternative to propane for in situ bioremediation of waste sites contaminated with VC.

Characterization of different biomasses based on their sugar profile with focus on their utilization for microbial biodiesel production.

The investigation and characterization of different biomasses as potential feedstocks for microbial lipid production was the aim of the present work. Twenty-eight biomasses were analyzed for total sugar content (glucose and xylose) and C/N-ratio as two major criteria for a suitable lipid production by oleaginous microorganisms. Considering both parameters, 9 out of 28 biomasses seemed to be suitable feedstocks for microbial lipid production. To allow for a broad evaluation of the suitability of the potential feedstocks, the estimated sugar and potential energy yield per area were calculated and the production costs and energy demand of the feedstock production were considered.

Draft Genome Sequence of Rhodococcus rhodochrous Strain ATCC 21198

ABSTRACT Rhodococcus rhodochrous is a Gram-positive red-pigmented bacterium commonly found in the soil. The draft genome sequence for R. rhodochrous strain ATCC 21198 is presented here to provide genetic data for a better understanding of its lipid-accumulating capabilities.

Effects of n-hexadecane and PM-100 clay on trichloroethylene degradation by Burkholderia cepacia

Trichloroethylene (TCE) is a non-flammable, volatile organochlorine compound which was a widely used degreasing agent, anesthetic, and coolant prior to 1960, but has since been placed on the Environmental Protection Agencys (EPA) list of priority pollutants. The inadequate disposal practices for TCE have created numerous TCE-contaminated superfund sites. The most commonly employed practice for remediating TCE-contaminated sites is to purge the contaminant from the source and trap it onto an adsorbent which is disposed of in a landfill or by incineration. This investigation was undertaken to evaluate the effectiveness of Burkholderia cepacia strain G4 (G4) to regenerate used sorbents by degrading TCE from the sorbent directly or indirectly. The results of this investigation showed that G4 was capable of reducing TCE attached to PM-100 clay but at significantly reduced rate due to the slow desorption rate. Conversely, it was shown that G4 was capable of degrading TCE dissolved in n-hexadecane at the same rate as systems without n-hexadecane present. The reduction in TCE degradation when the TCE is attached to the PM-100 clay could be overcome by solvent rinsing the TCE from the clay with subsequent removal of the TCE from the n-hexadecane by G4.