Reduction of acetone to isopropanol using producer gas fermenting microbes.

Gasification-fermentation is an emerging technology for the conversion of lignocellulosic materials into biofuels and specialty chemicals. For effective utilization of producer gas by fermenting bacteria, tar compounds produced in the gasification process are often removed by wet scrubbing techniques using acetone. In a preliminary study using biomass generated producer gas scrubbed with acetone, an accumulation of acetone and subsequent isopropanol production was observed. The effect of 2 g/L acetone concentrations in the fermentation media on growth and product distributions was studied with "Clostridium ragsdalei," also known as Clostridium strain P11 or P11, and Clostridium carboxidivorans P7 or P7. The reduction of acetone to isopropanol was possible with "C. ragsdalei," but not with P7. In P11 this reaction occurred rapidly when acetone was added in the acidogenic phase, but was 2.5 times slower when added in the solventogenic phase. Acetone at concentrations of 2 g/L did not affect the growth of P7, but ethanol increased by 41% and acetic acid concentrations decreased by 79%. In the fermentations using P11, growth was unaffected and ethanol concentrations increased by 55% when acetone was added in the acidogenic phase. Acetic acid concentrations increased by 19% in both the treatments where acetone was added. Our observations indicate that P11 has a secondary alcohol dehydrogenase that enables it to reduce acetone to isopropanol, while P7 lacks this enzyme. P11 offers an opportunity for biological production of isopropanol from acetone reduction in the presence of gaseous substrates (CO, CO2, and H2).

Process development for biological production of butanol from Eastern redcedar.

Eastern redcedar is an invasive softwood species in Oklahoma and across grasslands in the Central Plains of the United States and potential feedstock for butanol production. Butanol has higher energy content than ethanol and can be upgraded to jet and diesel fuels. The objective of this study was to develop a process for production of butanol from redcedar. Results showed that Clostridium acetobutylicum ATCC 824 and Clostridium beijerinckii NCIMB 8052 did not grow in fermentation medium with citrate buffer. However, both strains grew in the medium with acetate buffer, resulting in 3-4g/L greater butanol than without acetate. Detoxification of redcedar hydrolyzate was required to increase butanol concentration from 1 to 13g/L. Hydrolyzate was detoxified by activated carbon to remove inhibitors. Fermentations in detoxified redcedar hydrolyzate reached 13g/L butanol and 19g/L total ABE, comparable to glucose control. This shows the potential for redcedar use in butanol production.

Life cycle assessment of the production of ethanol from eastern redcedar.

This life cycle assessment (LCA) evaluates the environmental impacts of an ethanol production system using eastern redcedar (Juniperus virginiana L.) as the feedstock. Aspen Plus® was used to model the acid bisulfite pretreatment, enzymatic hydrolysis, fermentation, and distillation steps. A cradle-to-gate LCA was conducted to evaluate the environmental impacts from cutting the trees to the production of anhydrous ethanol. The environmental impacts of the redcedar ethanol process were compared to those from the production of corn ethanol. Inventory data for the system were collected and used to calculate a life cycle impact assessment (LCIA) using the IMPACT 2002+ and BEES+ framework in SimaPro 8.0.0. Four impact categories were evaluated: land occupation, water use, greenhouse gas (GHG) emissions, and non-renewable energy use. Results indicate that acid bisulfite pretreatment contributed to 65% of GHG emissions, 81% of non-renewable energy use, and 77% of water use of the overall process.

Simultaneous saccharification and fermentation of Eastern redcedar heartwood and sapwood using a novel size reduction technique.

This study investigated the effect of two wood zones (sapwood versus heartwood) and size reduction techniques [Crumbles® (Crumbles® is a registered trademark of Forest Concepts, LLC, Auburn, WA, USA) particles versus ground particles] on wood glucan-to-ethanol yield after acid bisulfite pretreatment and simultaneous saccharification and fermentation (SSF) of Eastern redcedar. SSFs were conducted at 8% solids loading (w/w dry basis) using Accellerase® 1500 at a loading of 46FPU/g glucan and Saccharomyces cerevisiae D5A for ethanol fermentation. The size reduction technique had no effect on ethanol yield. However, sapwood glucan-to-ethanol yields were significantly greater than heartwood yields. The highest wood glucan-to-ethanol yield of 187L/dryMg (95% of theoretical) was achieved with sapwood crumbled particles in 240h. Ground sapwood, crumbled heartwood and ground heartwood achieved ethanol yields of 89%, 81% and 80% of theoretical in 240h, respectively. Preliminary mass balances showed 100% glucan recovery with crumbled sapwood and extensive (72%) delignification.

Development of an efficient pretreatment process for enzymatic saccharification of Eastern redcedar.

This study investigates the potential for extracting sugars from the polysaccharides of Eastern redcedar. Pretreatment temperature, time, sulfuric acid loading, sodium bisulfite loading and impregnation time were varied using factorial treatment design experiments for identifying near optimal overall wood glucan-to-glucose yields during acid bisulfite pretreatments. The highest overall wood glucan-to-glucose yield of 87% was achieved when redcedar was impregnated with pretreatment liquor containing 3.75 g of sulfuric acid/100g of dry wood and 20 g of sodium bisulfite/100g of dry wood at 90 °C for 3h followed by increasing the temperature to 200 °C with a hold time of 10 min. Hemicellulose and lignin removal during pretreatments made the substrate amenable to enzymatic hydrolysis using 0.5 ml of Accelerase® 1500/g of glucan at 2% (w/w) solid loading. Preliminary mass balances showed 97% glucan recovery at pretreatment condition with 87% overall wood glucan-to-glucose yield and 59% delignification.

Effect of high dry solids loading on enzymatic hydrolysis of acid bisulfite pretreated Eastern redcedar.

This study investigates hydrolysis of cellulose from Eastern redcedar to glucose at high solids loading. Enzymatic hydrolysis of pretreated redcedar was performed with 0.5 ml Accelerase® 1500/g glucan (46 FPU/g glucan) using dry solids loading from 2% to 20% (w/w). Rheological challenges observed at high solids loading were overcome by adding stainless steel balls to shake flask reactors. The highest glucose concentration, 126 g/L (84% glucan-to-glucose yield), was obtained using 20% solids loading with stainless steel balls as a mixing aid. This enzymatic hydrolyzate was fermented into ethanol using Saccharomyces cerevisiae D5A to produce 52 g/L of ethanol (corresponding to 166 L/dry Mg of redcedar). Reducing enzyme dosage at 16% solids loading from 46 to 11.5 FPU/g glucan reduced glucan-to-glucose yields. This study has demonstrated the possibility of extracting sugars from the invasive species of Eastern redcedar with high solid loadings and their conversion into ethanol.

Effects of cellulose, hemicellulose and lignin on thermochemical conversion characteristics of the selected biomass.

The objective of this study was to investigate effects of biomass constituents (cellulose, hemicellulose and lignin) on biomass thermal decomposition and gas evolution profiles of four biomass materials. Switchgrass, wheat straw, eastern redcedar and dry distilled grains with solubles (DDGS) were selected as the biomass materials. No significant difference was observed in the weight loss profiles of switchgrass, wheat straw and eastern redcedar even though their cellulose, hemicellulose and lignin contents were considerably different. The weight loss kinetic parameters were also not significantly different except for activation energy of the eastern redcedar. However, biomass composition did significantly affect gas evolution profiles. The higher contents of cellulose and hemicellulose in switchgrass and wheat straw may have resulted in their higher CO and CO(2) concentrations as compared to eastern redcedar. On the other hand, higher lignin content in eastern redcedar may have resulted in significantly its high CH(4) concentration.