Keith Schimmel

Bioconversion of glycerol to ethanol by a mutant Enterobacter aerogenes

The main objective of this research is to develop, by adaptive evolution, mutant strains of Enterobacter aerogenes ATCC 13048 that are capable of withstanding high glycerol concentration as well as resisting ethanol-inhibition. The mutant will be used for high ethanol fermentation from glycerol feedstock. Ethanol production from pure (P-) and recovered (R-) glycerol using the stock was evaluated. A six-tube-subculture-generations method was used for developing the mutant. This involved subculturing the organism six consecutive times in tubes containing the same glycerol and ethanol concentrations at the same culture conditions. Then, the glycerol and/or ethanol concentration was increased and the six subculture generations were repeated. A strain capable of growing in 200 g/L glycerol and 30 g/L ethanol was obtained. The ability of this mutant, vis-à-vis the original strain, in utilizing glycerol in a high glycerol containing medium, with the concomitant ethanol yield, was assessed. Tryptic soy broth without dextrose (TSB) was used as the fermentation medium. Fermentation products were analyzed using HPLC.In a 20 g/L glycerol TSB, E. aerogenes ATCC 13048 converted 18.5 g/L P-glycerol and 17.8 g/L R-glycerol into 12 and 12.8 g/L ethanol, respectively. In a 50 g/L P-glycerol TSB, it utilized only 15.6 g/L glycerol; but the new strain used up 39 g/L, yielding 20 g/L ethanol after 120 h, an equivalence of 1.02 mol ethanol/mol-glycerol. This is the highest ethanol yield reported from glycerol bioconversion. The result of this P-glycerol fermentation can be duplicated using the R-glycerol from biodiesel production.

Effects of fertilizer application and dry/wet processing of Miscanthus x giganteus on bioethanol production.

The effects of wet and dry processing of miscanthus on bioethanol production using simultaneous saccharification and fermentation (SSF) process were investigated, with wet samples showing higher ethanol yields than dry samples. Miscanthus grown with no fertilizer, with fertilizer and with swine manure were sampled for analysis. Wet-fractionation was used to separate miscanthus into solid and liquid fractions. Dilute sulfuric acid pretreatment was employed and the SSF process was performed with saccharomyces cerevisiae and a cocktail of enzymes at 35°C. After pretreatment, cellulose compositions of biomass of the wet samples increased from 61.0-67.0% to 77.0-87.0%, which were higher than the compositions of dry samples. The highest theoretical ethanol yield of 88.0% was realized for wet processed pretreated miscanthus, grown with swine manure. Changes to the morphology and chemical composition of the biomass samples after pretreatment, such as crystallinity reduction, were observed using SEM and FTIR. These changes improved ethanol production.

Optimization of cultural conditions for conversion of glycerol to ethanol by Enterobacter aerogenes S012

The aim of this research is to optimize the cultural conditions for the conversion of glycerol to ethanol by Enterobacter aerogenes S012. Taguchi method was used to screen the cultural conditions based on their signal to noise ratio (SN). Temperature (°C), agitation speed (rpm) and time (h) were found to have the highest influence on both glycerol utilization and ethanol production by the organism while pH had the lowest. Full factorial design, statistical analysis, and regression model equation were used to optimize the selected cultural parameters for maximum ethanol production. The result showed that fermentation at 38°C and 200 rpm for 48 h would be ideal for the bacteria to produce maximum amount of ethanol from glycerol. At these optimum conditions, ethanol production, yield and productivity were 25.4 g/l, 0.53 g/l/h, and 1.12 mol/mol-glycerol, repectively. Ethanol production increased to 26.5 g/l while yield and productivity decreased to 1.04 mol/mol-glycerol and 0.37 g/l/h, respectively, after 72 h. Analysis of the fermentation products was performed using HPLC, while anaerobic condition was created by purging the fermentation vessel with nitrogen gas.

Effect of Viscosity on Membrane Fluxes in Cross-Flow Ultrafiltration

Abstract For practical applications of ultrafiltration (UF), an estimation of membrane fluxes under various operational conditions is very important. This study analyzed concentration polarization (CP) as a coupled transport problem with concentration-dependent solute viscosity. Besides the effects of variable viscosity, the model includes the effects of solute osmotic pressure, solute rejection at the membrane surface, and the axial pressure drop. This provides a fundamental understanding of the effects of various operating parameters on concentration polarization and transmembrane flux. A finite-difference solution of the transport equations is presented to model the concentration polarization in a thin-channel UF system. Simulation results for ultrafiltration of Dextran T-70 show that concentration-dependent solute viscosity adversely affects the transmembrane flux and needs to be carefully considered in modeling concentration polarization in membrane filtration.

Membrane Bioreactor Model for Removal of Organics from Wastewater

Abstract The persistence of trace organics in wastewater effluent is a major environmental concern. Possible use of fixed microbial films in wastewater treatment processes is currently an active area of research that may be able to address many of these problems. In the waste effluent, the persistence of trace organics is attributed, in part, to the inability of microbial populations to extract energy from dilute environments at a rate fast enough to sustain themselves. To address this problem, a novel wastewater treatment scheme is considered. On the basis of previous hollow fiber biomass growth studies, we believe that an anaerobic biofilm supported by hollow fibers could achieve greater biomass density than a film grown on traditional impermeable supports. This in turn could lead to improved substrate removal efficiency in a reactor of a given volume. Using this concept, we developed a mathematical model to test the potential of hollow fiber membrane reactors for biodegradation of acetate solution. A c...

Uses of miscanthus press juice within a green biorefinery platform.

This study assesses some uses of nutrient-rich juice mechanically extracted from freshly harvested Miscanthus x giganteus (MxG) as part of a green biorefinery system. The juice was used for culturing Saccharomyces cerevisiae and lactic acid bacteria. MxG juice was further used as substrate for fermentation to produce lactic acid using Lactobacillus brevis and Lactobacillus plantarum. The results show that MxG juice was a highly nutritious source for the cultivation of bacteria. Higher concentrations of MxG juice used as culture media, resulted in higher cell growth both aerobically and anaerobically. The highest ethanol yield of 70% theoretical and concentration of 0.75g/100ml were obtained from S. cerevisiae cultivated with 90% (v/v) MxG juice media and used for miscanthus solid fraction fermentation. 11.91g/L of lactic acid was also successfully produced from MxG juice through SSF.

Microarray analysis of the effect of Cowpea (Vigna unguiculata) phenolic extract in bovine peripheral blood

ABSTRACT In this study, the effect of polyphenolic extracts from cowpea (Vigna unguiculata) on global gene expression in bovine peripheral blood was investigated. Blood collected from Holstein-Friesian cows (n = 10) was treated with 10 µg/mL of cowpea phenolic extract (CPE) and subsequently used for transcriptional profiling using the Agilent bovine (v2) 4 × 44 K array. Calculation of fold change in gene expression and pathway analysis was conducted using the GeneSpring GX software 13.0. Real-time quantitative PCR was performed to validate the microarray data. Phenolic extracts of cowpea impacted global gene expression and resulted in 3170 differentially expressed genes (p < .05); 1716 genes were upregulated and 1454 genes were downregulated. Exposure to CPE impacted 66 pathways (p < .05) including the Wnt signalling pathway, Toll-like receptor pathway, inflammation response pathway, MAPK cascade pathway, prostaglandin synthesis and regulation pathway, cell cycle pathway, insulin signalling pathway, and the adipogenesis pathway. Expression of immune markers such as CD40, CD68, Toll-like receptors, and Wnt signalling changed. Exposure to CPE modulated expression of genes associated with immunity and homeostasis. Transcriptional profiles of the response to polyphenols may aid in the design of targeted diets to influence animal production and health and thus requires further study.

SCALING IN SITU BIOREMEDIATION PROBLEMS BY APPLICATION OF MULTIPHASE, MULTICOMPONENT TRANSPORT THEORY

A methodology for scaling in situ bioremediation problems is presented. This methodology is based on multiphase, muiticomponent transport theory and employs inspectional analysis and numerical sensitivity studies. A general mathematical model that describes subsurface aerobic biotransformation of organic chemical species in a multiphase setting is first presented. This general model is applied to the specific case of microbial enhanced vapor-vacuum extraction (MEVVE) in a one-dimensional zone of immobile liquids. The resulting simplified MEVVE model considers rate-dependent interphase mass transfer, a flowing gaseous phase, a single hydrocarbon pseudo-component, and either oxygen or hydrocarbon limited microbial activity. By inspectional analysis a set of dimensionless groups are derived that represent the various model parameters. A scries of numerical sensitivity studies are presented that examine the impact of selected dimensionless groups on overall system biotransformation rates. This analysis demons...