Progress 09/01/10 to 08/31/12
Outputs
OUTPUTS: This project continued to develop new processing technologies to underpin second-generation biorefineries using soybeans and corn as feedstocks to produce biofuels (biodiesel and bioethanol) and replace imported petroleum. Hexane is highly flammable posing major safety concerns and is a regulated pollutant for which compliance with new emission standards is increasingly difficult. Aqueous extraction processing (AEP) using water to extract oil is inherently safe and suitable for small-scale plants. Enzyme-assisted AEP (EAEP) integrated well into biodiesel and corn ethanol plants and synergies may make conversion of both crops into fuels, biobased products, food and feed more cost-effective. We developed and tested new processing strategies using water and enzymes to process soybeans into edible oil and value-added protein products. These technologies may help meet the biofuel targets of the 2007 Energy Independence and Security Act. The project also sought to break the food-vs-fuel dogma by producing improved and healthy food and enhanced feed ingredients as biorefinery co-products. We demonstrated proof-of-concept for integrating EAEP into corn grain-based biofuel production. We developed a spectrum of products to give corn-soybean biorefineries flexibility to produce an optimum array of products that maximizes profits. We demonstrated that it is highly desirable to integrate our EAEP process into a corn/soybean biorefinery to improve feed products, conserve water and enzyme use, produce "organic" oil, increase ethanol fermentation rate, and utilize byproducts to increase ethanol yield. These "bolt-on" technologies should make dry-grind corn fuel ethanol plants much more cost-effective and reduce tensions in the "food vs fuel" debate. EAEP fiber-rich feed products were evaluated for proximate analysis, digestibility, and amino acid quality but had limited value in poultry diets, other species may be able to better utilize high-fiber components. We developed solid-state fermentation of fiber streams to produce healthy lipids containing long-chain polyunsaturated fatty acids. We worked with BIO-CAT, Inc., to develop applications for protein and sugar streams as a plant growth promoter. Field-plot studies demonstrated clear benefits of applying hydrolyzed soy protein as part of a growth promoter package. We partnered with Modular Genetics to integrate biosurfactant production (dispersant for oil spill cleanup) with our soybean biorefinery concept and evaluated hulls and the liquid (skim) and solid (fiber) byproduct streams as nutrients for surfactant production. We provided surfactant samples to collaborators to evaluate surface activity and toxicity to marine species. We published 15 peer-reviewed articles in scientific journals and made 8 presentations at scientific conferences. We trained scientists and engineers on non-thermal, non-chemical processing and membrane technology in food systems, at the Central Institute of Agricultural Engineering, Bophal, India. Two MS and one PhD graduate students, and one graduate intern, were trained. We provided research training to 6 postdoctoral research associates and one visiting scientist from Brazil. 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, scaling up demulsification, and fermentation of soybean fiber; 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; and Michael E. Persia, Assistant Professor, Animal Science, responsible for animal feeding trials. The following Iowa State University staff worked on the project: Devin Maurer, Research Associate; Catherine Hauck, Research Associate; William Colonna, Research Associate; Hui Wang, CCUR Pilot Plant Manager; and Show-ling Lee, Research Scientist. The following graduate students and post-doctoral research associates received training experiences by working on the project: 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; graduate intern, Clement Augereau from France; Juliana M.L. Nobrega de Moura, Post-doctoral Research Associate; Virginie Kapchie, Post-doctoral Research Associate; Shengli Yang, Post-doctoral Research Associate; Mustafa Marti, Post-doctoral Research Associate; and Linxing Yao, Post-doctoral Research Associate. Professor Neiva de Almeida, Visiting Research Scientist from Federal University of Paraiba (Brazil) worked on the project. Six undergraduate students also worked on the project gaining laboratory experiences. Collaborators and Contacts: We collaborated with Christopher Penet of BIO-CAT, Inc. (Troy, VA) on developing applications for the protein and sugar fractions as plant growth promoters and with Kevin Jarrell, Modular Genetics (Woburn, MA) on fermenting soybean fractions to produce biosurfactants. TARGET AUDIENCES: Target audiences include: soybean and corn farmers and soybean and corn grower's associations, such as the American Soybean Association, the United Soybean Board, and National Corn Growers Association; Feed Grains Council; renewable fuels associations; National Biodiesel Board; soybean and corn processing industry, such the National Oilseed Processors Association and Corn Refiners Association. PROJECT MODIFICATIONS: Co-principle investigator P.A. Murphy retired in May 2011, but completed all of her work plan tasks. No other major changes were made.
Impacts
EAEP of soybeans is an environmentally friendly process to produce oil for food and biofuels. Nearly all oil can be extracted from soybean solids, but the oil unclaimed in skim limits oil yield, and protein/sugar fractions have been unmarketable products. We developed solutions to these problems that integrate with corn ethanol production. We optimized nanofiltration of soy whey produced by isoelectric precipitation of protein from EAEP skim so that edible soy protein isolate can be produced, which is important to capturing value from the protein. Nanofiltration was used to concentrate the whey and ultrafiltration to filter the skim fraction from which permeate was used to slurry corn for fermentation. Good protein yields and concentration factors were achieved. We optimized corn fermentation using concentrated whey to improve ethanol yield and the nutritional quality of the feed product distillers dry grains with solubles (DDGS). We developed an enzyme process to convert soy oligosaccharides into fermentable sugars. We integrated soybean EAEP with corn ethanol fermentation at 100 and 50% skim replacement. The 100% as-is skim replacement of water in corn fermentation gave the best fermentation and feed quality. Hydrolyzing soybean oligosaccharides improved fermentation rate for skim and whey fractions. All EAEP fractions increased the protein content of DDGS, but using skim to slurry corn for ethanol fermentation gave the highest protein content in DDGS. We scaled-up aqueous ammonia soaking pretreatment and an improved enzyme treatment to convert soy fiber into fermentable sugars. When fermenting with S. cerevisiae, ethanol concentration was low. Sequential fermentation with hexose-converting S. cerevisiae and pentose- and hexose-converting E. coli KO11 of the enzyme-pretreated fiber increased ethanol production without chemical pretreatment. We determined the energy and amino acid digestibility for soy-corn DDGS and soy fiber fractions using roosters. We improved the pilot-plant procedure for a second process in which oil is recovered as intact oil bodies (oleosome), and demonstrated its reproducibility in achieving high oil yield. Most soy storage protein was recovered by washing oleosomes. The soy oleosomes were characterized for their protein composition electrical charges, particle size, and stabilities against creaming and oxidation. Free oil recovery from oleosomes was scaled up in the pilot plant. The soy-corn DDGS was used as substrate in solid-state fermentation with three fungi combinations and optimized conditions. Xylanase and cellulase activities reduced fiber content. The soy-DDGS was evaluated in chick feeding and rooster digestibility trials. Corn ethanol fermentation using the nanofiltered soy skim had the same rate-promoting effect on yeast growth and ethanol fermentation as whole skim. Carbohydrase-treated soy hulls were viable replacements for glucose in producing the biosurfactant. Arachidonic and eicosapentaenoic acids are extremely important human nutrients and were produced by Pythium irregulare fungus fermentation using soybean cotyledon fiber and soy skim as substrates. A corn-soybean biorefinery is viable.
Publications
- 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.
- Towa, L.T., Kapchie, V.N., Hauck, C.C., Wang, H., and Murphy, P.A. 2011. Pilot Plant Recovery of Soybean Oleosome Fractions by an Enzyme-assisted Aqueous Process. J. Amer. Oil Chem. Soc. 88:733-741.
- Towa, L.T., Kapchie, V.N., Wang, H., Hauck, C., Wang, T., and Murphy, P.A. 2011. Quantity and Quality of Free Oil Recovered from Enzymatically Disrupted Soybean Oleosomes. J. Amer. Oil Chem. Soc. 88:1581-1591.
- Kapchie, V.N., Hauck, C., Wang, H., and Murphy, P.A. 2011. Process Improvement for Semipurified Oleosomes on a Pilot-Plant Scale. J. Food Sci. 76:C853-C860.
- Karki, B., Maurer, D., Kim, H., and Jung, S. 2012. Ethanol Production from Soybean Fiber, a Co-product of Soybean Oil Extraction, Using Aqueous Ammonia Soaking, J. Am. Oil Chem. Soc., DOI 10.1007/s11746-012-2016-z.
- 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. Am. Oil Chem. Soc. 88:1649-1658.
- 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. Am. Oil Chem. Soc. 88:1045-1051.
- Karki, B., Maurer, D., and Jung, S. 2011. Efficiency of Pretreatments for Optimal Enzymatic Saccharification of Soybean Insoluble Fractions. Bioresource Technol. 102:6522-6528.
- Jung, S., de Moura, J.M.L.N., Campbell, K., and Johnson, L.A. 2011. Enzyme-assisted Aqueous Extraction of Oilseeds. In Enhancing Extraction Processes in the Food Industry, edited by N. Lebovka, E. Vorobiev, and F. Chemat, Contemporary Food Engineering, Series Editor-in-Chief Da-Wen Sun, CRC Press LLC, Taylor and Francis Group, Boca Raton, FL.
- Campbell, K.A., Glatz, C.E., Johnson, L.A., Jung, S., de Moura, J.M.N., Kapchie, V., and Murphy, P. 2011. Advances in Aqueous Extraction Processing of Soybeans. J. Am. Oil Chem. Soc. 88:449-465.
- Karki, B., Maurer, D., Kim, T.H., and Jung, S. 2011. Comparison and Optimization of Enzymatic Saccharification of Soybean Fibers Recovered from Aqueous Extractions. Bioresource Technol. 102:1228-1233.
- Lio, J.Y., and Wang, T. 2012. Solid-state Fermentation of Soybean and Corn Processing Co-products for Improved Feed Quality. J. Agric. Food Chem. 60:7702-7709.
- Yao, L., Lee, S., Wang, T., de Moura, J.M.L.N., and Johnson, L.A. 2012. Effects of Fermentation Substrate Conditions on Corn-soy Co-fermentation for Fuel Ethanol Production. Bioresource Technol. 120:140-148.
- 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.
- Ndlela, S.C., de Moura, J.M.L.N., Olson, N.K., and Johnson, L.A. 2012. Aqueous Extraction of Oil and Protein from Soybeans with Subcritical Water. J. Am. Oil Chem. Soc. 89(6):1145-1153.
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Progress 09/01/10 to 08/31/11
Outputs
OUTPUTS: This project continues our efforts to develop new processing technologies to underpin second-generation biorefineries using soybeans and corn as feedstocks to produce biofuels (biodiesel and bioethanol), replacing imported petroleum. Hexane is highly flammable posing major safety concerns and is a regulated pollutant for which compliance with new emission standards is increasingly difficult. Aqueous extraction processing (AEP) using water is inherently safe and suitable for small-scale plants. Enzyme-assisted AEP (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. We developed and tested new processing strategies using water and enzymes to process soybeans into edible oil and value-added protein products. 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 demonstrated proof-of-concept at laboratory scale for integrating a water- and enzyme-based approach known as Enzyme-assisted Aqueous Extraction Processing (EAEP) into corn grain-based biofuel production. We seek to develop a spectrum of products to give corn-soybean biorefineries flexibility to produce an optimum array of products that maximizes profits. 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. These "bolt-on" technologies should make dry-grind corn fuel ethanol plants much more cost effective and reduce tensions in the "food vs fuel" debate. We worked with BIO-CAT, Inc., to develop applications for protein and sugar streams as a plant growth promoter. Sufficient amounts of soy protein fractions were produced to complete growth study. The field-plot study demonstrated clear benefits of applying hydrolyzed soy protein as part of growth promoter package. We partnered with Modular Genetics to integrate biosurfactant production (dispersant for oil spill cleanup) with a soybean biorefinery and provided samples of surfactant to external collaborators to evaluate surface activity and toxicity to marine species. The materials were prepared by fermentation and purification procedures jointly developed with our industrial collaborator. We published 4 peer-reviewed articles in scientific journals and made 3 presentations at scientific conferences (including Turkey and India). We trained scientists and engineers on non-thermal, non-chemical processing and membrane technology in food systems, at the Central Institute of Agricultural Engineering, Bophal, India. Two MS graduate students, one graduate intern, and one PhD graduate student were trained. We provided research training to five postdoctoral research associates and one visiting scientist from Brazil. 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; and Michael Persia, Assistant Professor, Animal Science, responsible for animal feeding trials. The following Iowa State University staff worked on the project: Devin Maurer, Research Associate; Catherine Hauck, Research Associate; William Colonna, Research Associate; Hui Wang, CCUR Pilot Plant Manager; and Show-ling Lee, Research Scientist. The following graduate students and post-doctoral research associates received training experiences by working on the project: 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; graduate intern, Clement Augereau from France; Juliana M.L. Nobrega de Moura, Post-doctoral Research Associate; Virginie Kapchie, Post-doctoral Research Associate; Shengli Yang, Post-doctoral Research Associate; Mustafa Marti, Post-doctoral Research Associate; and Linxing Yao, Post-doctoral Research Associate. Professor Neiva de Almeida, Visiting Research Scientist from Federal University of Paraiba (Brazil) worked on the project. Six undergraduate students also worked on the project gaining laboratory experiences. Collaborators and Contacts: We collaborated with Christopher Penet of BIO-CAT, Inc. (Troy, VA) on developing applications for the protein and sugar fractions as plant growth promoters and with Kevin Jarrell, Modular Genetics (Woburn, MA) on fermenting soybean fractions to produce biosurfactants. TARGET AUDIENCES: Target audiences include: soybean and corn farmers and soybean and corn grower's associations, such as the American Soybean Association, the United Soybean Board, and National Corn Growers Association; Feed Grains Council; renewable fuels associations; National Biodiesel Board; soybean and corn processing industry, such as the National Oilseed Processors Association and Corn Refiners Association. PROJECT MODIFICATIONS: Co-principal investigator P.A. Murphy retired in May 2011, but has completed all of her work plan tasks. No other major changes were made.
Impacts
Water-based EAEP of soybeans is an environmentally friendly process to produce oil for food and biofuels. Nearly all oil can be extracted from soybean solids using EAEP, but the amount unclaimed from skim limits oil recovery and protein/sugar fractions are unmarketable products. We developed several approaches to solve these problems and integrate with corn ethanol production. We optimized nanofiltration of soy whey produced by isoelectric precipitation of protein from EAEP skim so that edible soy protein isolate can be produced, which is important to capturing value from the protein. Oil, protein, and solids mass balances were determined so that economic assessments can be made. Nanofiltration was used to concentrate the whey and ultrafiltration to filter the skim fraction from which permeate was used to slurry corn for fermentation. Good protein yields and concentration factors were achieved. We optimized the use of concentrated whey in corn fermentation to improve ethanol yield and nutritional quality of the feed product distillers dry grains with solubles (DDGS). We developed an enzyme process using galactosidase to convert soy oligosaccharides into fermentable sugars. We scaled-up aqueous ammonia soaking pretreatment and an improved enzyme treatment to convert soy fiber into fermentable sugars. When fermenting with S. cerevisiae, ethanol concentration was low. Sequential fermentation with hexose-converting S. cerevisiae and pentose- and hexose-converting E. coli KO11 of the enzyme-pretreated fiber increased ethanol production chemical pretreatment. We are determining the energy and amino acid digestibility for enhanced soy-corn DDGS and soy fiber fractions using roosters. We improved the pilot-plant procedure for a second process in which oil is recovered as intact oil bodies (oleosome), and demonstrated its reproducibility in achieving high oil yield. Most soy storage protein was recovered by washing oleosomes. Semi-purified and purified soy oleosomes were characterized for their protein composition electrical charges, particle size, and stabilities against creaming and oxidation. Free oil recovery from oleosomes was scaled up in the pilot plant. We integrated soybean EAEP with corn ethanol fermentation with 100 and 50% skim replacement. The soy-corn DDGS was used as substrate for large-scale solid-state fermentation using 3 fungi combinations and conditions we optimized. High xylanase and cellulase activities and reduced fiber content were achieved. The soy-DDGS are being evaluated in chick and rooster feeding trials. Corn ethanol fermentation using the nanofiltered soy skim had the same rate-promoting effect on yeast growth and ethanol fermentation as whole skim. The 100% as-is skim replacement of water in corn fermentation gave the best fermentation and feed quality. Modular Genetics has based discussions with potential customers on the results of our fermentations to produce biosurfactants on soy byproducts applied for additional funding to extend the project to additional alternative biosurfactants and to scale-up of the process to produce sufficient amounts for commercial evaluation.
Publications
- Towa, L.T., Kapchie, V.N., Hauck, C.C., Wang, H. and Murphy, P.A. 2011. Pilot Plant Recovery of Soybean Oleosome Fractions by an Enzyme-assisted Aqueous Process. J. Amer. Oil Chem. Soc. 88:733-741.
- Campbell, K.A., Glatz, C.E., Johnson, L.A., Jung, S., de Moura, J.M.N., Kapchie, V. and Murphy, P. 2011. Advances in Aqueous Extraction Processing of Soybeans. J. Amer. Oil Chem. Soc. 88(4):449-465.
- Towa, L.T., Kapchie, V.N., Wang, G., Hauck, C., Wang, T., and Murphy, P.A. 2011. Quantity and Quality of Free Oil Recovered from Enzymatically Disrupted Soybean Oleosomes. J. Amer. Oil Chem. Soc. 88:1581-1591.
- Kapchie, V.N., Hauck, C., Wang, H., and Murphy, P.A. 2011. Process Improvement for Semipurified Oleosomes on a Pilot-Plant Scale. J. Food Sci. 76:C853-C860.
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