Progress 09/01/09 to 08/31/11
Outputs
OUTPUTS: This project continues an effort focused on developing new processing technologies to underpin second-generation biorefineries using soybeans and corn as feedstocks to produce biofuels (biodiesel and bioethanol) and biobased products (adhesives), replacing imported petroleum. We seek to integrate a water- and enzyme-based approach known as Enzyme-assisted Aqueous Extraction Processing (EAEP) into biofuel production replacing a hazardous and polluting petroleum-based solvent, hexane, used to extract vegetable oil. Hexane is a regulated pollutant and compliance with new emission standards is increasingly difficult. Hexane is also highly flammable posing major safety concerns and can be used only at very large scale. EAEP is inherently safe and suitable for small-scale plants. EAEP integrates well into biodiesel and corn ethanol production and synergies achieved may make conversion of both crops into fuels, biobased products, food and feed much more cost effective. These advanced technologies may help meet the biofuel targets of the 2007 Energy Independence and Security Act. The project also seeks to break the dogma of food verses fuel by producing improved and healthy food and enhanced feed ingredients as biorefinery co-products. We continued our partnership with Genencor International (enzyme manufacturer). We demonstrated at lab scale that it is highly desirable to integrate our EAEP process into a corn/soybean biorefinery to improve feed products, conserve water and enzyme use, eliminate hexane and produce "organic" oil, increase ethanol fermentation rate, and to utilize low value byproducts to increase ethanol yield. This "bolt-on" technology should make dry-grind fuel ethanol plants much more cost effective and reduce tensions in the "food vs fuel" debate. We also brought in a new partner in enzyme applications, BIO-CAT, Inc., to help us develop applications for protein and sugar streams. Sufficient amounts of soy protein fractions were produced to enable a soybean growth study. The field plot study demonstrated clear benefits of applying hydrolyzed soy protein as part of growth promoter package. Genencor continued to work with equipment manufacturers to improve adoption potential by industry. We demonstrated in the laboratory that EAEP can be integrated into a dry-grind ethanol production to achieve much improved processing efficiencies. ISU faculty published 13 peer-reviewed articles in scientific journals and made 12 presentations at scientific conferences (even in India, Ireland and Portugal where related research is underway). We presented our results at numerous public forums, such as the dedication of the ISU BioCentury Research Farm (a new prototype biorefinery) and the ISU Presidential Lecture Series to which the general public was invited. We graduated 2 PhD students and 1 MS student who worked on the project. Another 2 MS graduate students are being trained on the project. We provided research training to 2 postdoctoral research associates and 1 visiting scientist from Brazil. Two senior scientists from the Central Institute of Agricultural Engineering (India) were also trained on EAEP. PARTICIPANTS: Individuals (Principle Investigators): Lawrence A. Johnson, Project Director, Professor, Food Science & Human Nutrition, responsible for integrating and scaling up oil demulsification with 2-stage EAEP extraction and characterizing protein products; Charles E. Glatz, Professor, Chemical & Biological Engineering, responsible for evaluating alternative protein recovery strategies and comparing process alternatives; Stephanie Jung, Associate Professor, Food Science & Human Nutrition, responsible for integrating soy fiber conversion to ethanol production and scaling up demulsification; Patricia A. Murphy, University Professor, Food Science & Human Nutrition, responsible for enhancing oleosome extraction and recovering protein products; Tong Wang, Associate Professor, Food Science & Human Nutrition, responsible for integrating soy skim utilization in dry-grind ethanol production. The following Iowa State University staff worked on the project: Devin Maurer, Research Associate; Catherine Hauck, Research Associate; William Colonna, Research Associate. The following graduate students and post-doctoral research associates received training experiences by working on the project: Kerry Campbell, Post-doctoral Research Associate, Chemical & Biological Engineering; Bishnu Karki, PhD Student, Food Science & Human Nutrition; Shannon Box, MS Student, Food Science & Human Nutrition (minority student funding was acquired from ISU); Jun Yi Lio, MS student, Food Science & Human Nutrition; Juliana M.L. Nobrega de Moura, Post-doctoral Research Associate; Virginie Kapchie, Post-doctoral Research Associate; Shengli Yang, Post-doctoral Research Associate; Neiva de Almeida, Visiting Research Scientist from Federal University of Paraiba (Brazil); and Dipika Agrahar Murugkar, Visiting Senior Scientist from Central Institute of Agricultural Engineering (India). Six undergraduate students also worked on the project gaining laboratory experiences. Collaborators and Contacts: The team collaborated with the international enzyme manufacturer Genencor International (Rochester, NY) and several key Genencor International researchers contributed to the project's success, including Peter Birschbach, Jeff Gerstner, and Chris Barnett. We also collaborated with Christopher Penet of BIO-CAT, Inc. (Troy, VA) on developing applications for the protein and sugar fractions as plant growth promoters. TARGET AUDIENCES: Target audiences include: soybean farmers and soybean grower's associations, such as the American Soybean Association and the United Soybean Board; Feed Grains Council; renewable fuels associations; National Biodiesel Board; soybean processing industry, such the National Oilseed Processors Association. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
Water-based EAEP of soybeans is an environmentally friendly alternative to traditional hexane extraction to produce oil for food and biofuels. Nearly all oil can be extracted from soybean solids using EAEP, but the amount unclaimed from skim after separating the oil-rich cream limits oil recovery and protein/sugar fractions are challenging to convert to marketable products. We scaled up approaches designed to solve these limitations and challenges and demonstrated proof-of-concept for integrating EAEP with corn ethanol production. Two-stage countercurrent EAEP was successfully scaled-up to pilot-plant scale. Extraction and cream demulsification were integrated to enable enzyme recycling thereby conserving enzyme and reducing cost. Nearly complete (98%) oil and protein extractions were achieved. Enzymatic demulsification also worked well recovering nearly all oil in the cream. Overall oil recovery was approximately 76% due to 19% of the oil retained by the skim. Failure to recover more oil in the cream was due to a fraction of the emulsified oil binding to protein to a level where the particles are denser than the skim; thus they do not float. Membrane filtration enabled recovery of valuable protein products and over 90% of the protein was retained when using three different approaches with dead-end and cross-flow membrane filtration. When using isoelectric precipitation alone, protein recovery was inversely related to the extent of hydrolysis, with recoveries ranging from 27 to 87% of skim proteins. When coupling isoelectric precipitation with membrane filtration, more than 95% of the protein was recovered. Recovery of protein products requires matching separation technology to the degree of hydrolysis during extraction. Isoelectric precipitation recovered minimally hydrolyzed proteins with purity limited only by oil content. Protein functionality was affected by extent of hydrolysis with greater hydrolysis enhancing protein solubility, rate of foaming, and foaming stability but reducing emulsification properties. Essential amino acid compositions and in vitro protein digestibilities were not adversely affected by either extrusion or extraction treatments. Lunasin, a healthy bioactive peptide, was slightly reduced. Healthy isoflavones distributed between the skim and protein precipitates with most in the skim. Oleosome fractionation was successfully replicated on pilot-plant scale with improved oil yields and less protein proteolysis compared to lab scale. Oil release from oleosomes was accomplished with proteases and mechanical processing. The functionality of proteins from oleosome processing was characterized as well as phytochemicals in the oleosome process fractions. We developed an alternative cream demulsification process using methanol to integrate with biodiesel production. We discovered EAEP soy fiber is an excellent substrate for solid-state fermentation. Aspergillus oryzae, Trichoderma reesei, and Phanerochaete chrysosporium grew well and produced significant cellulase and xylanase activities. T. reesei achieved the greatest fiber degradation. Soy fiber, when added to distillers dry grains with solubles enhanced feed quality.
Publications
- Jung, S., de Moura, J.M.L.N., Campbell, K.A. and Johnson, L.A. 2011. Enzyme-assisted Aqueous Extraction of Oilseeds. In: Enhancing Extraction Processes in the Food Industry. Edited by Nikolai Lebovka, Eugene Vorobiev, and Farid Chemat, Series on "Contemporary Food Engineering", Taylor & Francis, 477-519.
- de Moura, J.M.L.N., Maurer, D., Jung, S. and Johnson, L.A. 2011. Pilot-plant Proof-of-Concept for Countercurrent Two-stage Enzyme-assisted Aqueous Extraction Processing of Soybeans. J. Amer. Oil Chem. Soc. 88:649-1658.
- de Moura, J.M.L.N., Maurer, D., Jung, S. and Johnson, L.A. 2011. Integrated Countercurrent Two-stage Extraction and Cream Demulsification in Enzyme-assisted Aqueous Extraction of Soybeans. J. Amer. Oil Chem. Soc. 88:1045-1051.
- Campbell, K.E. and Glatz, C.E. 2010. Protein Recovery from Enzyme-Assisted Aqueous Extraction of Soybean. Biotechnol. Prog. 26:488-495.
- Bishnu, K., Maurer, D., Kim, T. and Jung, S. 2010. Comparison and Optimization of Enzymatic Saccharification of Soybean Fibers Recovered from Aqueous Extractions. Bioresource Technol., doi:10.1016/j.biortech.2010.08.004.
- Kapchie, V.N., Towa, L.T., Hauck, C.C. and Murphy, P.A. 2010. Recycling of Aqueous Supernatants in Soybean Oleosomes Isolation. J. Amer. Oil. Chem. Soc. 87:223-231.
- Towa, L.T., Kapchie, V.N., Hauck, C.C. and Murphy, P.A. 2010. Enzyme Assisted Aqueous-extraction of Oil from Isolated Oleosomes of Soybean Flour. J. Amer. Oil Chem. Soc. 87:347-354.
- Yao, L, Wang, T. and Wang, H. 2011. Effect of Soy Skim from Soybean Aqueous Processing on the Performance of Corn Ethanol Fermentation. Bioresource Technology, doi:10.1016/j.biortech.2011.06.071.
- de Moura, J.M.L.N., Hernandez-Ledesma, B., de Almeida, N.M., Hsieh, C., de Lumen, B.O. and Johnson, L.A. 2011. Lunasin and Bowman-Birk Protease Inhibitor Concentrations of Protein Extracts from Enzyme-assisted Aqueous Extraction of Soybeans. J. Agric. Food Chem. 59:6940-6946.
- de Moura, J.M.L.N., Campbell, K., de Almeida, N.M., Glatz, C.E. and Johnson, L.A. 2011. Protein Extraction and Membrane Recovery in Enzyme-assisted Aqueous Extraction Processing of Soybeans. J. Am. Oil Chem. Soc. 88(6):877-889.
- de Moura, J.M.L.N., Maurer, D., Jung, S. and Johnson, L.A. 2011. Integrated Countercurrent Two-stage Extraction and Cream Demulsification in Enzyme-assisted Aqueous Extraction of Soybeans. J. Am. Oil Chem. Soc. 88(7):1045-1051.
- de Moura, J.M.L.N., Campbell, K., de Almeida, N.M. Glatz, C.E. and Johnson, L.A. 2011. Protein Recovery in Aqueous Extraction Processing of Soybeans Using Isoelectric Precipitation and Ultrafiltration. J. Am. Oil Chem. Soc. 88(9):1447-1454.
- Nobrega de Moura, J.M.L, de Almeida, N.M., Jung, S. and Johnson, L.A. 2010. Flaking as a Pretreatment for Enzyme-assisted Aqueous Extraction Processing of Soybeans. J. Am. Oil Chem. Soc. 87(12):1507-1515.
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Progress 09/01/09 to 08/31/10
Outputs
OUTPUTS: This project continues an effort focused on developing new processing technologies to underpin second-generation biorefineries using soybeans and corn as feedstocks to produce biofuels (biodiesel and bioethanol) and biobased products (adhesives), replacing imported petroleum. We seek to integrate a water- and enzyme-based approach known as Enzyme-assisted Aqueous Extraction Processing (EAEP) into biofuel production replacing a hazardous and polluting petroleum-based solvent, hexane, used to extract vegetable oil. Hexane is a regulated pollutant and compliance with new emission standards is increasingly difficult. Hexane is also highly flammable posing major safety concerns and can be used only at very large scale. EAEP is inherently safe and suitable for small-scale plants. EAEP integrates well into biodiesel and corn ethanol production and synergies achieved may make conversion of both crops into fuels, biobased products, food and feed much more cost effective. These advanced technologies may help meet the biofuel targets of the 2007 Energy Independence and Security Act. The project also seeks to break the dogma of food verses fuel by producing improved and healthy food and enhanced feed ingredients as biorefinery co-products. We continued our partnership with Genencor International (enzyme manufacturer). We also brought in a new partner in enzyme applications, BIO-CAT, Inc., to help us develop applications for protein and sugar streams. Genencor continued to work with equipment manufacturers to improve adoption potential by industry. We demonstrated in the laboratory that EAEP can be integrated into a dry-grind ethanol production to achieve much improved processing efficiencies. ISU faculty published 3 peer-reviewed articles in scientific journals and made 12 presentations at scientific conferences (even in India, Ireland and Portugal where related research is underway). We presented our results at numerous public forums, such as the dedication of the ISU BioCentury Research Farm (a new prototype biorefinery) and the ISU Presidential Lecture Series to which the general public was invited. We graduated 2 PhD students and 1 MS student who worked on the project. Another 2 MS graduate students are being trained on the project. We provided research training to 2 postdoctoral research associates and one visiting scientist from Brazil. Two senior scientists from the Central Institute of Agricultural Engineering (India) were also trained on EAEP. PARTICIPANTS: Individuals (Principle Investigators): Lawrence A. Johnson, Project Director, Professor, Food Science & Human Nutrition, responsible for integrating and scaling up oil demulsification with 2-stage EAEP extraction and characterizing protein products; Charles E. Glatz, Professor, Chemical & Biological Engineering, responsible for evaluating alternative protein recovery strategies and comparing process alternatives; Stephanie Jung, Associate Professor, Food Science & Human Nutrition, responsible for integrating soy fiber conversion to ethanol production and scaling up demulsification; Patricia A. Murphy, University Professor, Food Science & Human Nutrition, responsible for enhancing oleosome extraction and recovering protein products; Tong Wang, Associate Professor, Food Science & Human Nutrition, responsible for integrating soy skim utilization in dry-grind ethanol production. The following Iowa State University staff worked on the project: Devin Maurer, Research Associate; Catherine Hauck, Research Associate; William Colonna, Research Associate. The following graduate students and post-doctoral research associates received training experiences by working on the project: Kerry Campbell, PhD Candidate, Chemical & Biological Engineering; Yating Ma, MS Student, Food Science & Human Nutrition; Bishnu Karki, PhD Student, Food Science & Human Nutrition; Shannon Box, MS Student, Food Science & Human Nutrition (minority student funding was acquired from ISU); Jun Yi Lio, MS student, Food Science & Human Nutrition; Juliana M.L. Nobrega de Moura, Post-doctoral Research Associate; Post-doctoral Research Associate; Virginie Kapchie, Post-doctoral Research Associate; Shengli Yang, Post-doctoral Research Associate; Neiva de Almeida, Visiting Research Scientist from Federal University of Paraiba (Brazil); and Dipika Agrahar Murugkar, Visiting Senior Scientist from Central Institute of Agricultural Engineering (India). Six undergraduate students also worked on the project gaining laboratory experiences. Collaborators and Contacts: The team collaborated with the international enzyme manufacturer Genencor International (Rochester, NY) and several key Genencor International researchers contributed to the project's success, including Peter Birschbach, Jeff Gerstner, and Chris Barnett. We also collaborated with Christopher Penet of BIO-CAT, Inc. (Troy, VA) on developing applications for the protein and sugar fractions as plant growth promoters. TARGET AUDIENCES: Target audiences include: soybean farmers and soybean grower's associations, such as the American Soybean Association and the United Soybean Board; Feed Grains Council; renewable fuels associations; National Biodiesel Board; soybean processing industry, such the National Oilseed Processors Association. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
Water-based EAEP of soybeans is an environmentally friendly alternative to traditional hexane extraction to produce oil for food and biofuels. Nearly all oil can be extracted from soybean solids using EAEP, but the amount unclaimed from skim after separating the oil-rich cream limits oil recovery and protein/sugar fractions are challenging to convert to marketable products. We scaled up approaches designed to solve these limitations and challenges and demonstrated proof-of-concept for integrating EAEP with corn ethanol production. Two-stage countercurrent EAEP was successfully scaled-up to pilot-plant scale. Extraction and cream demulsification were integrated to enable enzyme recycling thereby conserving enzyme and reducing cost. Nearly complete (98%) oil and protein extractions were achieved. Enzymatic demulsification also worked well recovering nearly all oil in the cream. Overall oil recovery was approximately 76% due to 19% of the oil retained by the skim. Failure to recover more oil in the cream was due to a fraction of the emulsified oil binding to protein to a level where the particles are denser than the skim; thus they do not float. Membrane filtration enabled recovery of valuable protein products and over 90% of the protein was retained when using three different approaches with dead-end and cross-flow membrane filtration. When using isoelectric precipitation alone, protein recovery was inversely related to the extent of hydrolysis, with recoveries ranging from 27 to 87% of skim proteins. When coupling isoelectric precipitation with membrane filtration, more than 95% of the protein was recovered. Recovery of protein products requires matching separation technology to the degree of hydrolysis during extraction. Isoelectric precipitation recovered minimally hydrolyzed proteins with purity limited only by oil content. Protein functionality was affected by extent of hydrolysis with greater hydrolysis enhancing protein solubility, rate of foaming, and foaming stability but reducing emulsification properties. Essential amino acid compositions and in vitro protein digestibilities were not adversely affected by either extrusion or extraction treatments. Lunasin, a healthy bioactive peptide, was slightly reduced. Healthy isoflavones distributed between the skim and protein precipitates with most in the skim. Oleosome fractionation was successfully replicated on pilot-plant scale with improved oil yields and less protein proteolysis compared to lab scale. Oil release from oleosomes was accomplished with proteases and mechanical processing. The functionality of proteins from oleosome processing was characterized as well as phytochemicals in the oleosome process fractions. We developed an alternative cream demulsification process using methanol to integrate with biodiesel production. We discovered EAEP soy fiber is an excellent substrate for solid-state fermentation. Aspergillus oryzae, Trichoderma reesei, and Phanerochaete chrysosporium grew well and produced significant cellulase and xylanase activities. T. reesei achieved the greatest fiber degradation. Soy fiber, when added to distillers dry grains with solubles enhanced feed quality.
Publications
- Bishnu, K., D. Maurer, T. Kim, and S. Jung. 2010. Comparison and Optimization of Enzymatic Saccharification of Soybean Fibers Recovered from Aqueous Extractions. Bioresource Technol., doi:10.1016/j.biortech.2010.08.004.
- Kapchie, V.N., L.T. Towa, C.C. Hauck, and P.A. Murphy. 2010. Recycling of Aqueous Supernatants in Soybean Oleosomes Isolation. J. Amer. Oil. Chem. Soc. 87:223-231.
- Towa, L.T., V.N. Kapchie, C.C. Hauck, and P.A. Murphy. 2010. Enzyme Assisted Aqueous-extraction of Oil from Isolated Oleosomes of Soybean Flour. J. Amer. Oil Chem. Soc. 87:347-354. DOI 10.1007/s11746-009-1503-3.
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