Progress 09/01/10 to 08/31/14

Outputs
Target Audience:The target audiences for this research effort are three fold. First the biofuels industry is very enthusiastic for the research shown here in order for the development of an alternative fuel source using a variety of biomass in the forest. Secondly, the United States Forest Service is a significant target audience if alternative substrates from overcrowded forests are used for this process. The rural economic development will be an advantage, but if material thinned from our forests can be used then the health of the forests and help in prevention of forest fires. Third, the United States Department of Energy is the last target audience, as this technology will enhance our resource base by providing new carbohydrate for fermentation. Each new identified source of carbohydrate that can be converted into fuels and chemicals will help to increase our energy independence. Changes/Problems:Pilot scale success was paramount to the commercial success of the technology. This stepwas slower than anticipated due to mechanical and engineering complications with our pilot scale facility, which required some substantial modifications in order to properly run our substrate through the first stage of processing. We were finally able to get good process control in order to run the substrates. We were also successful by processing the liquid and solid materials for our fermentation scientists as well as the lignin sulfonates that we have supplied as well to our industrial partner. The results from our industrial partner helpedin modifications we will needed to make with the lignosulfate production processing conditions. What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? A collaborative effort between BioPulping International, the USDA Forest Products Laboratory, and the University of Wisconsin have developed a novel pretreatment using the SPORL process for robust conversion of woody biomass to fermentable sugars through enzymatic hydrolysis for bioethanol production. The SPORL process consists of reacting wood chips with a solution of sodium sulfite at elevated temperatures for a short time frames and then defiberizing using a disk refiner to generate fibrous substrate for subsequent saccharification and fermentation. The work involves FPL, UW-Madison, and Catchlight Energy, a Chevron Company. Presently ethanol is made where grain is available. The product from our pretreatment is a sugar resource that can be converted into ethanol or other fermentation products. An ethanol manufacturing facility could be located with a SPORL plant. Locating new ethanol production in areas where there is limited grain will allow for locally produced ethanol to be incorporated into transportation fuels with less expense. Additional impact can be derived if material from overcrowded forests is used for this process. The rural economic development will be an advantage, but if material thinned from our forests can be used then the health of the forests and help in prevention of forest fires will also be increased. This will enhance our resource base by providing new carbohydrate for fermentation and health of the environment by improving our standing forests. Each new identified source of carbohydrate that can be converted into fuels and chemicals will help to increase our energy independence. Our studies indicate that monomeric sugar recovery from spruce wood after a SPORL pretreatment contained a significant amount of glucose from dissolved cellulose by the pretreatment. Considering hemicellulose content of about 23 percent in spruce, monomer sugar recovery rate from hemicellulose is estimated to be 80 percent. Preliminary fermentation of the pretreatment spent liquor found that it can be easily fermented to produce good ethanol yield. Major efforts were made on using the CD300 laboratory pilot scale mechanical pulping system to conduct one step SPORL pretreatment, i.e., conducting chemical pretreatment of wood chips and post pretreatment wood size reduction through disk milling in one step. We also made effort to achieve mass balance. We have developed an approach to account for biomass mass balance. In this approach, we will collect everything from wood chip pre-squeeze, pretreatment hydrolysate from squeeze of the pretreated wood chips by the screw plug to transfer to the disk mill, and the total liquid and solid at the cyclone discharge. Then we will analyze the chemical composition of each collected stream. Our results show that SED of over 70 percent can be achieved for Aspen after 40 minutes pretreatment at nominal cellulase loading. Under similar SPORL pretreatment conditions and same cellulase loading, SED of approximately 90 percent was achieved at lab bench scale studies. Good news is that recently, through the course of this project, we have generated some interesting findings as a new invention that could lead to intellectual property. The invention has three aspects, 1 is the developed appropriate enzymatic hydrolysis conditions, specifically the range of pH, to significantly increase lignocelluloses enzymatic saccharification efficiency, especially for substrates whose lignin contains sulfonate groups; 2 is a new and novel use of lignin that contains sulfonate groups including lignosulfonate as an additive to enhance enzymatic hydrolysis of lignocelluloses. When applying this invention to sulfite based pretreatments of lignocellulosic biomass for bioconversion, one can use pretreatment hydrolysate (hemicellulosic sugar stream) that contains lignosulfonate to achieve robust simultaneous enzymatic saccharification and combined fermentation (SSCombF) of enzymatic and pretreatment hydrolysates. Finally 3 is a novel process that combines 1 and 2, conduct SSComb at the proper enzymatic hydrolysis conditions using pretreatment hydrolysates that contain lignosulfonate, including any lignin with sulfonate groups. This is especially applicable to SPORL and SO2 catalyzed pretreatments with or without explosion. The benefits of this invention are, eliminate the solid lignocellulosic substrate washing to reduce water usage, approximately 5000 ton/day for a typical biorefinery of 1000 ton biomass/day, in biorefineries through SSCombF and reduce cellulase enzyme loading by as much as 46 percent by using the optimized pH and lignosulfonate.

Publications

• Type: Journal Articles Status: Published Year Published: 2011 Citation: J.Y. Zhu, R. Gleisner, S. Tian, J. Negron, E. Horn, and X. Luo (2011). Efficient Ethanol Production from Beetle-Killed Lodgepole Pine using SPORL Technology and Saccharomyces cerevisiae without Detoxification. TAPPI Journal, May 1, 2011.

Progress 09/01/11 to 08/31/12

Outputs
OUTPUTS: The use of the SPORL process for robust conversion of woody biomass to fermentable sugars through enzymatic hydrolysis for bioethanol production has had several outputs. First, several new techniques has been developed with experimentation throughout this project using a variety of temperatures and chemical loading. Second, several visiting scientists were able to work on the project, which allowed them to contribute to the project objectives, but also receive some learning, through mentoring, about wood physics and biotechnology in the pulp and paper and bioethanol industry. Third, the successful development of this technology has enhanced existing bio-processing through collaborations with industry that see a need for making new sources of materials available that can be used for a fermentation resource for local ethanol production in areas that have little available grain production. Finally, the extracted sugar hyrolysates have been shared with a number of research institutions for evaluation of fermentative ability. These evaluations through conducting and analyzing experiments have assisted researchers and students to better understand the nature of the hydrolysates and the best organisms for optimal ethanol production. The collaborative efforts of the Forest Products Laboratory and the University of Wisconsin have significantly helped to progress this project as well as contributing to this novel technology. PARTICIPANTS: Forest Products Laboratory collaborators include Dr. Jun Yong Zhu, Research Engineer, Dr. Shen Tian, Research Microbiologist, Wenyuan Zhu, Scientific Technician, and Roland Gleisner, General Engineer. Other participants to the SPORL experiments were Dr. Xuejun Pan, Biochemist, from the University of Wisconsin-Madison. All scientists and technicians were able to provide training and mentoring to visiting scientists. BioPulping International contributions include Dr. Masood Akhtar, CEO, and PI of the project. Dr. Akhtar was crucial in experimental planning and data analysis. Finally, Eric Horn, Research Microbiologist for BPI, provided technical expertise for the pretreatments, sugar extractions, fiberization, sample collection, and sample facilitation. TARGET AUDIENCES: The target audiences for this research effort are three fold. First the biofuels industry is very enthusiastic for the research shown here in order for the development of an alternative fuel source using a variety of biomass in the forest. Secondly, the United States Forest Service is a significant target audience if alternative substrates from overcrowded forests are used for this process. The rural economic development will be an advantage, but if material thinned from our forests can be used then the health of the forests and help in prevention of forest fires. Third, the United States Department of Energy is the last target audience, as this technology will enhance our resource base by providing new carbohydrate for fermentation. Each new identified source of carbohydrate that can be converted into fuels and chemicals will help to increase our energy independence. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
SPORL processing is unique in that the products from our pretreatment is a sugar resource that can be converted into ethanol or other fermentation products. Locating new ethanol production in areas where there is limited grain will allow for locally produced ethanol to be incorporated into transportation fuels with less expense. Additional impact can be derived if material from overcrowded forests is used for this process. Using thinned material from our forests will increase the health and help in prevention of forest fires as well as enhancing our resource base by providing new carbohydrate for fuel and overall health of the environment. Each new identified source of carbohydrate that can be converted into fuels and chemicals will help to increase our energy independence. Our studies indicate that monomeric sugar recovery from spruce wood after a SPORL pretreatment contained a significant amount of glucose from dissolved cellulose by the pretreatment. Considering hemicellulose content of about 23 percent in spruce, monomer sugar recovery rate from hemicellulose is estimated to be 80 percent. Preliminary fermentation of the pretreatment spent liquor can be easily fermented to produce good ethanol yield. Major efforts were made on using the CD300 laboratory pilot scale mechanical pulping system to conduct one step SPORL pretreatment, i.e., conducting chemical pretreatment of wood chips and post pretreatment wood size reduction through disk milling in one step. We also made effort to achieve mass balance. We have developed an approach to account for biomass mass balance. In this approach, we will collect everything from wood chip pre-squeeze, pretreatment hydrolysate from squeeze of the pretreated wood chips by the screw plug to transfer to the disk mill, and the total liquid and solid at the cyclone discharge. Then we will analyze the chemical composition of each collected stream. Our results show that SED of over 70 percent can be achieved for Aspen after 40 minutes pretreatment at nominal cellulase loading. Under similar SPORL pretreatment conditions and same cellulase loading, SED of approximately 90 percent was achieved at lab bench scale studies. Good news is that recently, through the course of this project, we have generated some interesting findings as a new invention that could lead to intellectual property. The invention has three aspects. The first is the developed appropriate enzymatic hydrolysis conditions, specifically the range of pH, to significantly increase lignocelluloses enzymatic saccharification efficiency, especially for substrates whose lignin contains sulfonate groups. The second is a new and novel use of lignin that contains sulfonate groups including lignosulfonate as an additive to enhance enzymatic hydrolysis of lignocelluloses. The third is a novel process that combines 1 and 2, conduct SSComb at the proper enzymatic hydrolysis conditions using pretreatment hydrolysates that contain lignosulfonate, including any lignin with sulfonate groups. The benefits of this invention are eliminating the solid lignocellulosic substrate washing to reduce water usage and reduce cellulase enzyme loading by 46 percent.

Publications

• J.Y. Zhu, R. Gleisner, S. Tian, J. Negron, E. Horn, and X. Luo 2011. Efficient Ethanol Production from Beetle-Killed Lodgepole Pine using SPORL Technology and Saccharomyces cerevisiae without Detoxification. TAPPI Journal, May 1, 2011.

Progress 09/01/10 to 08/31/11

Outputs
OUTPUTS: The use of the SPORL process for robust conversion of woody biomass to fermentable sugars through enzymatic hydrolysis for bioethanol production has had several outputs. First, several new techniques has been developed with experimentation throughout this project using a variety of temperatures and chemical loading. Second, several visiting scientists were able to work on the project, which allowed them to contribute to the project objectives, but also receive some learning, through mentoring, about wood physics and biotechnology in the pulp and paper and bioethanol industry. Third, the successful development of this technology has enhanced existing bio-processing through collaborations with industry that see a need for making new sources of materials available that can be used for a fermentation resource for local ethanol production in areas that have little available grain production. Finally, the extracted sugar hyrolysates have been shared with a number of research institutions for evaluation of fermentative ability. These evaluations through conducting and analyzing experiments have assisted researchers and students to better understand the nature of the hydrolysates and the best organisms for optimal ethanol production. The collaborative efforts of the Forest Products Laboratory and the University of Wisconsin have significantly helped to progress this project as well as contributing to this novel technology. PARTICIPANTS: Forest Products Laboratory collaborators include Dr. Jun Yong Zhu, Research Engineer, Dr. Shen Tian, Research Microbiologist, Wenyuan Zhu, Scientific Technician, and Roland Gleisner, General Engineer. Other participants to the SPORL experiments were Dr. Xuejun Pan, Biochemist, from the University of Wisconsin-Madison. All scientists and technicians were able to provide training and mentoring to visiting scientists. BioPulping International contributions include Dr. Masood Akhtar, CEO, and PI of the project. Dr. Akhtar was crucial in experimental planning and data analysis. Finally, Eric Horn, Research Microbiologist for BPI, provided technical expertise for the pretreatments, sugar extractions, fiberization, sample collection, and sample facilitation. TARGET AUDIENCES: The target audiences for this research effort are three fold. First the biofuels industry is very enthusiastic for the research shown here in order for the development of an alternative fuel source using a variety of biomass in the forest. Secondly, the United States Forest Service is a significant target audience if alternative substrates from overcrowded forests are used for this process. The rural economic development will be an advantage, but if material thinned from our forests can be used then the health of the forests and help in prevention of forest fires. Third, the United States Department of Energy is the last target audience, as this technology will enhance our resource base by providing new carbohydrate for fermentation. Each new identified source of carbohydrate that can be converted into fuels and chemicals will help to increase our energy independence. PROJECT MODIFICATIONS: No major project modifications reported

Impacts
A collaborative effort between BioPulping International, the USDA Forest Products Laboratory, and the University of Wisconsin have developed a novel pretreatment using the SPORL process for robust conversion of woody biomass to fermentable sugars through enzymatic hydrolysis for bioethanol production. The SPORL process consists of reacting wood chips with a solution of sodium sulfite at elevated temperatures for a short time frames and then defiberizing using a disk refiner to generate fibrous substrate for subsequent saccharification and fermentation. The work involves FPL, UW-Madison, and Catchlight Energy, a Chevron Company. Presently ethanol is made where grain is available. The product from our pretreatment is a sugar resource that can be converted into ethanol or other fermentation products. An ethanol manufacturing facility could be located with a SPORL plant. Locating new ethanol production in areas where there is limited grain will allow for locally produced ethanol to be incorporated into transportation fuels with less expense. Additional impact can be derived if material from overcrowded forests is used for this process. The rural economic development will be an advantage, but if material thinned from our forests can be used then the health of the forests and help in prevention of forest fires will also be increased. This will enhance our resource base by providing new carbohydrate for fermentation and health of the environment by improving our standing forests. Each new identified source of carbohydrate that can be converted into fuels and chemicals will help to increase our energy independence. Our studies indicate that monomeric sugar recovery from spruce wood after a SPORL pretreatment contained a significant amount of glucose from dissolved cellulose by the pretreatment. Considering hemicellulose content of about 23 percent in spruce, monomer sugar recovery rate from hemicellulose is estimated to be 80 percent. Preliminary fermentation of the pretreatment spent liquor found that it can be easily fermented to produce good ethanol yield. Major efforts were made on using the CD300 laboratory pilot scale mechanical pulping system to conduct one step SPORL pretreatment, i.e., conducting chemical pretreatment of wood chips and post pretreatment wood size reduction through disk milling in one step. We also made effort to achieve mass balance. We have developed an approach to account for biomass mass balance. In this approach, we will collect everything from wood chip pre-squeeze, pretreatment hydrolysate from squeeze of the pretreated wood chips by the screw plug to transfer to the disk mill, and the total liquid and solid at the cyclone discharge. Then we will analyze the chemical composition of each collected stream. Our results show that SED of over 70 percent can be achieved for Aspen after 40 minutes pretreatment at nominal cellulase loading. Under similar SPORL pretreatment conditions and same cellulase loading, SED of approximately 90 percent was achieved at lab bench scale studies.

Publications

• J.Y. Zhu, R. Gleisner, S. Tian, J. Negron, E. Horn, and X. Luo (2011). Efficient Ethanol Production from Beetle-Killed Lodgepole Pine using SPORL Technology and Saccharomyces cerevisiae without Detoxification. TAPPI Journal, May 1, 2011.