2008 Protein and Co-Products
PCP 1: Non-Food Utilization of Animal Protein By-Products
Chair(s): G. Piazza, USDA, ARS, ERRC, USA; and R. Garcia, USDA, ARS, ERRC, USA
Industrial Protein Utilization: The Multifunctioning "By-Product". J. Schmitz, Iowa State University, Ames, IA, USA
The term green products has a variety of meanings, but most commonly it refers to biofuels and clean energy. Biobased products are viewed as the future of materials, but an inspection of the past shows they were prevalent through much of the 20th century. The current resurgence in biobased products has to some extent bypassed protein sources - animal and plant sources alike. A detailed overview of nonfood and nonfeed protein use is presented along with thoughts on the future scale of protein sources that may be available.
Novel Peptide-Based Materials from Restructured Agricultural Proteins. J. Barone, A. Athamneh, N. Budhavaram, Z. Li, Virginia Polytechnic Institute and State University, Blacksburg, VA, USA
The ratio of ordered to disordered (O/D) regions in a protein determines its physical properties. This is reminiscent of the naïve “fringed micelle” model in polymer science. It is possible to change the O/D ratio through several methods, including external plasticization, internal plasticization, and enzymatic re-structuring. External plasticization involves using a small molecular weight polar molecule to lower protein Tg and increase strain to break (ε) by increasing the amount of disordered regions in the protein. Internal plasticization is a functionalization technique that increases ε and lowers Tg by covalently attaching bulky side groups (BSG) to the protein to increase disordered regions. Ordered regions in the protein can be increased by enzymatically cleaving a protein at a BSG that is a known structure-breaker then re-attaching the peptide segments in a close-packed array with another enzyme.
Biodegradable Plastics from Animal Protein Co-Products. S. Sharma, J.N. Hodges, I. Luzinov, School of Materials Science and Engineering, Clemson University, Clemson, SC, USA
Recently, the outbreak of Bovine Spongiform Encephalopathy (BSE) in Europe has led to prohibition/limitation of the use of various animal co-product proteins in ruminant feed. The excessive availability of these protein materials has encouraged the search for alternative uses of them, such as fabrication of biodegradable plastics. In this research work, plastic samples from partially denatured feathermeal and bloodmeal proteins were successfully produced by the compression-molding process. The modulus (stiffness) for the material obtained was found to be comparable with that of commercial synthetic material but with lower toughness characteristics, which is a common phenomenon among plastics produced from animal and plant proteins. Several ways to improve the properties of the plastics have been explored.
Production of Peptones from Animal Protein By-Products for Use in Industrial Fermentation. R.A. Garcia1, D. Pyle2, G.J. Piazza1, Z. Wen2, 1Fats Oils and Animal Coproducts Research Unit, Eastern Regional Research Center, Agricultural Research Service, Wyndmoor, PA, USA, 2Department of Biological Systems Engineering, Virginia Polytechnic Institute and State University, Blacksburg, VA, USA
Animal protein by-products such as meat & bone meal, hydrolyzed feather meal, or blood meal may be useful as a source of nitrogen and micronutrients for industrial fermentations, especially for fermentations involving fastidious microorganisms that do not grow well in chemically-defined media. These by-products, however, are not well suited for use in fermentation media without further processing. In the present work, animal by-products were defatted, hydrolyzed by enzyme or alkali treatment and then spray dried. Hydrolysates of different protein by-products, hydrolyzed in various manners, were analyzed for degree of hydrolysis and the degradation, racemization and crosslinking of amino acids. Practical performance characteristics including the viscosity, foaming, autoclave stability and clarity of hydrolysate solutions were also determined. The overall results indicate that enzymatic hydrolysis can produce a superior product, but at greater cost. Choice of a hydrolysate method ultimately depends on the requirements of the specific fermentation organism.
Meat and Bone Meal Extract and Gelatin as Renewable Flocculants. G.J. Piazza, R.A. Garcia, Eastern Regional Research Center, ARS, USDA, Wyndmoor, PA, USA
We investigated whether proteins can be used as renewable clay flocculants to potentially replace polyacrylamide (PAM), a flocculant derived from natural gas. A laboratory test for clay flocculation was developed. This test was validated using commercial anionic PAM. When calcium chloride was present, anionic PAM promoted clay flocculation, but PAM was not effective without calcium chloride. Two soy proteins, a whey fraction, a porcine gelatin, and a meat & bone meal (MBM) extract were used in the flocculation test. It was found that MBM extract and porcine gelatin promoted clay flocculation. With the addition of calcium chloride, 88, 99, and 97% of the clay was settled after 24 h in the presence of optimal amounts of PAM, gelatin, and MBM extract, respectively. Without the addition of calcium chloride, gelatin and MBM extract were still effective, and 99 and 97% of the clay was settled after 24 h in the presence of optimal amounts of these proteins, respectively. These figures compare favorably to those from no flocculant controls, in which approximately 50% of the clay had settled with or without the addition of calcium chloride after 24 h. However, compared to PAM more than 20 times more gelatin and more than 500 times more MBM extract were needed for optimal clay flocculation. Also analysis showed that the MBM extract contained mostly degraded protein.
Potential Uses of a Novel Mucin Extracted From Jellyfish. K. Ushida1, T. Baba1, M. Urai1, K. Taniguchi1, J. Uzawa1, K. Kihira1, A. Masuda2, N. Dohmae2, M. Yamamura3, H. Wada3, 1Eco-Soft Materials Research Unit, Riken, Wako, Saitama, Japan, 2Biomolecular Characterization Team, Riken, Wako, Saitama, Japan, 3Shinwa Chemical Industries, Ltd., Kyoto, Japan
Mass occurrence of jellyfish is now called “Jellyfish Explosion” and reported all around the world probably because of the effect of marine pollutions, global warming, and overfishing. A novel mucin was extracted from various kinds of jellyfish which accumulate in a huge amount on fishing sites and power plants as marine wastes. Its main chain structure made of short tandem repeats with 8 amino acids and the core structure around O-glycoside bonds were clarified. The amino acid sequence of tandem repeat –VVETTAAP- is similar to that of human mucin MUC5AC, -TTSTTSAP-. Since the jellyfish is a primitive animal species. this mucin has very simple glycoforms and can be used as a starting material to produce a designer mucin by use of glycosyltransferases. It has a large potential to be used in various application including medical use, pharmacy, food additives, and cosmetics because it can be a base material to make artificial mucus mimicking its functions that are indispensable to sustain lives. The approximate yield of mucin is about 1 kg from several tons of jellyfish in wet weight. The mass production of mucin from wastes with simple and well-defined structure are now in our scope.
Investigating Protein Hydrolysates for Use as Herbivore Repellents. B.A. Kimball1,2, 1USDA, APHIS, National Wildlife Research Center, Philadelphia, PA, USA, 2Monell Chemical Senses Center, Philadelphia, PA, USA
Damage to agricultural, horticultural, and ornamental plants by foraging herbivores is a significant and costly problem. Among the many non-toxic active ingredients that have been tested as repellents, animal products such as egg, blood and urine have consistently proven to be the most efficacious. It was previously thought that malodorous volatiles released from these animal-based repellents invoked a ′fear response′ through their chemosensory association with predators. However, this mechanism is inconsistent with observed herbivore behavior. I propose that the protein fraction of animal-based repellents deters herbivory by altering palatability of the treated food. Accordingly, several proteins and protein hydrolysates were evaluated as herbivore repellents. In experiments with captive deer (Odocoileus hemionus columbianus), casein hydrolysate-treated foods were completely avoided. Casein hydrolysate also reduced intake of mountain beavers (Aplodontia rufa) and pocket gophers (Thomomys mazama), whereas rabbits (Oryctolagus spp.) demonstrated greater avoidance of hydrolyzed collagen. Subsequent studies have demonstrated that species adopting strict herbivorous digestive strategies are more likely to avoid protein hydrolysates than omnivorous species such as rats (Ratus spp.) or coyotes (Canis latrans).
Gasification of Salmon Processing Waste. C. DeWitt1, S. Rowland1, K. Patil1, C. Bower2, 1Oklahoma State University, Stillwater, OK, USA, 2USDA, ARS, Fairbanks, AK, USA
The seasonal salmon harvest is often conducted in isolated areas in Alaska. Infrastructure is often lacking and traditional waste remediation strategies, such as rendering, often are not economically feasible. An alternative approach was therefore sought to capture value from salmon waste. Gasification is a process that uses high temperatures (700°C) in a low oxygen environment to produce gas (syngas). This project attempts to determine whether high moisture products, such as salmon waste, can be gasified. Production of syngas from high moisture products is difficult. A drying agent is therefore needed to reduce moisture. A good source of dry material is municipal solid waste (i.e. paper, corrugate, and wood pallets). Therefore, salmon waste products were dried to 20% moisture content using wood pellets. Products evaluated were whole fish, heads, viscera, or frames (raw or de-oiled). Syngas heating value was measured from H2, CO, CH4, C2H2, C2H4, and C2H6 produced from biomass during gasification. Results demonstrated that the heating value of salmon waste mixed with pellets was not significantly different from whole pellets. Efficiency of gas production in the pilot scale gasifier, however, was only about 25% when compared to maximum values obtained using bomb calorimetery. Results demonstrate gasification is possible when salmon biomass is dried using wood pellets.
Preserving High-Protein Fish By-Products through Silages and Fermentates. C.K. Bower, K.A. Hietala, USDA , ARS, Fairbanks, AK, USA
In Alaska, over three million metric tons of fish by-products are generated each year. However, due to the remote locations and seasonal nature of salmon fisheries, by-products are generally not fully utilized unless a fish meal plant is located nearby. Acidification is a common method for inhibiting microorganisms and promoting autolysis of animal tissues, thereby preserving perishable foods such as fish.In these studies, salmon by-products were stabilized using a variety of acidification methods, then the quality of the high-protein fish was evaluated. Hot-smoking technologies reduced the pH to 4.8, preserved proteins, and eliminated all “fishy” odors. Fermentation by lactic acid bacteria lowered the salmon by-products to pH 4.5 during 18 weeks of storage, but destroyed much of the fish protein. Fermentations using local agricultural discards such as potatoes as a carbohydrate source were not effective for stabilizing the pH of salmon. Ensilage through direct acidification using organic and inorganic acids decreased protein quality, but maintained a pH below 4.5 for 18 weeks. Alternative methods of preservation are needed to decrease the loss of valuable marine proteins and oils, and to provide salmon processors with environmentally sound options for adding value to by-products currently discarded as waste.
PCP 2: Co-Product Utilization from Biofuels Processing
Chair(s): K. Liu, USDA, ARS, USA; and T. Tran, Solae, USA
Recent Development in Ethanol Processes and Co-Product Composition. J. Shetty1, M. Hruby2, 1Genencor-Danisco, Palo Alto, CA, USA, 2Danisco Animal Nutrition, St. Louis, MO, USA
The rapid growth of fuel alcohol primarily from corn necessitates the need for greater fermentation efficiencies, reduced energy cost and higher value co-products. This presentation provides a discussion on the different processes such as raw starch hydrolysis (STARGEN Process), Phytase Amylase Liquefaction Process (PALS Process) dry and wet corn fractionation processes for ethanol production and its impact on the composition and value of co-products.One such co-product, corn dried distillers grains with solubles (DDGS), is present at levels of up to over 20% in poultry and pig feeds. DDGS from PALS process has shown significant reduction in phytate phosphorus and improvement in energy and amino acid digestibility. Producing DDGS, which is highly desirable by animal feed producers can offer additional market differentiation to ethanol producers.
Novel Food Ingredients from Bio-Ethanol. T. Lohrmann, D. Hammes, Quality Technology International, Inc., Elgin, IL, USA
Today the primary method for the manufacture of ethanol from corn involves grinding the entire kernel for fermentation. The residual, non-fermented co-product from this process is termed distillers grains (DG). A soluble fraction of nutrients remaining after fermentation is added during the drying process. The resulting product is called dried distillers grains with solubles (DDGS). DDGS is not an ideal food substrate or ingredient and thus is sold into the livestock feed industry. A new modified wet milling ethanol pre-processing system has been developed that allows for the efficient separation of corn grain, in a food-grade facility, into 3 basic fractions: germ, bran and endosperm prior to fermentation. While the endosperm fraction is used for ethanol, the germ and fiber are separated out early in the process. Since this is a wet fractionation system, the corn fractions are cleanly and efficiently separated into high purity food ingredients. A unique aspect of this new wet fractionation system is that it does not use sulfur dioxide as part of the process. The germ and fiber fractions from this process are new and unique substrates that have been shown to have novel benefits for the food industry. From these fractions high value corn oil, protein isolates and fiber have been processed.
Characterization of the Oil, Protein, and Carbohydrate Fractions Generated by an Enzyme-Assisted Nonsolvent Oilseed Extraction Process. Peter N. Birschbach, Genencor - A Danisco Division, Rochester, NY, USA
There is a growing demand for oilseed extraction processes that do not use organic solvents such as hexane. In collaboration with Iowa State University, Genencor has developed an enzyme-assisted nonsolvent process for extraction of oil from oilseeds which we presented at the 2007 AOCS Annual Meeting. In the past year we have made several significant improvements in the process itself enabling us to reduce water consumption, reduce drying costs, improve free oil recovery, and operate the process in a semi-continuous mode. We have also done significant analytical and applications studies to establish the value of the fractions generated by our process. The enzyme-assisted nonsolvent process generates a protein-rich skim fraction, a carbohydrate-rich insoluble fraction, and a free oil fraction. The free oil fraction is suitable for use in food applications or for conversion to biodiesel. The skim fraction contains a high concentration of non-denatured proteins which are suitable for use in a wide range of human and animal nutrition applications. The carbohydrate-rich insoluble fraction is suitable for use in animal nutrition or as a feedstock for bioethanol production. We will present analytical and applications data to support these conclusions.
Review of Fiber Separation From Distillers Dried Grains with Solubles (DDGS) Using Sieving and Air Classification, and Utilization of the Fiber to Produce Valuable Products. R. Srinivasan, Mississippi State University, Mississippi State University, MS, USA
A combination of two separation methods, sieving and elutriation (air flow); called the "Elusieve process", was developed to separate fiber from distillers dried grains with solubles (DDGS) in the dry grind corn process. Two products were obtained by separating fiber: 1) enhanced DDGS with lower fiber, higher fat and higher protein contents and 2) fiber. DDGS samples were sieved into five sieve categories. The two smallest sieve categories comprised > 40% of the original DDGS weight and had lower fiber and higher protein and fat contents. Elutriation of the larger three sieve categories resulted in higher protein and fat contents and lower fiber contents in the fractions that settled at the bottom of the elutriation column. Fiber in DDGS had higher particle density than nonfiber. Fiber was flat shaped while nonfiber was nearly spherical. Fiber particles were carried selectively at low air velocity despite higher density because: 1) fiber particles in the sieve category have lower or comparable weight to smallest nonfiber particles due to their flatness and 2) flat shaped particles experience higher drag force. Enhanced DDGS had higher phytosterol content than Elusieve fiber. Elusieve fiber was suitable for production of corn fiber gum and ethanol.
Fractionation of Distiller's Dry Grains with Solubles (DDGS) by Sieveing and Winnowing. Keshun Liu, United States Dept. of Agriculture, Agricultural Research Services, Aberdeen, ID, USA
Four commercial distiller's dried grains with solubles (DDGS) were subjected to sieving. All sieved fractions except for the pan fraction, constituting about 90% of original mass, were subjected to winnowing with an air blast seed cleaner. Sieving was effective in producing fractions with varying composition. As the mesh size increased, protein and ash contents increased, total carbohydrate (CHO) decreased, and oil change varied with samples. Winnowing sieved fractions was also effective. Protein and oil were enriched in heavy sub-fractions, while CHO enriched in light sub-fractions. As the velocity increased, light sub-fraction mass increased, protein, oil and ash contents increased but CHO decreased in both heavy and light sub-fractions, and the compositional difference between the two decreased. For protein, there was a balanced effect between sieving and winnowing, resulting a maximum 56.4% reduction in a fraction and maximum 60.2% increase in another fraction. Winnowing was more effective than sieving in changing oil and CHO contents. Winnowing whole sample was not as effective as sieving winnowed fractions in changing composition, but their combined effect appeared comparable to that of sieving followed by winnowing. Fractionating DDGS not only enhances nutritional values but also expands market shares.
Study of Lipids and Lipid Components in Corn-Dried Distiller's Grains. J.K. Winkler, K.A. Rennick, F.J. Eller, S.F. Vaughn, USDA, ARS, NCAUR, USA
Corn distiller's dried grain (DDG) is a major by-product of ethanol fermentation from corn processed by dry-milling, and is high in fiber, protein, and oil. DDG is sold primarily for livestock feed and is currently valued at approximately $85-110 per ton. In efforts to increase the profitability of the corn-to-ethanol process, and to diversify the number of co-products, companies are starting to extract the valuable oil either before or after fermentation. The current emphasis is that this oil might then be used for biodiesel production. However, the oil extracted from DDG is significantly different from commercial corn oil, which is oil extracted from corn germ. Although the fatty acid composition is similar to corn germ oil, oil from DDG is higher in valuable food nutraceuticals including tocopherols, tocotrienols, phytosterols, and ferulate phytosterol esters. Methods such as molecular distillation as well as silica chromatography were used to isolate these valuable components from the oil. The ferulate phytosterol esters from DDG oil have similar antioxidant properties as oryzanol from rice bran oil, and have the added benefit of increased stability due to their unique composition. These phytochemicals may present yet another valuable co-product from ethanol fermentation
Soybean Flour and Isolate Hydrolysates for Adhesive Utilization. J. Schmitz1, D. Myers2, Y. Bian1, 1Iowa State University, Ames, IA, USA, 2North Dakota State University, Fargo, ND, USA
Soy adhesives was the primary natural adhesives for plywood production before 1960. Increasing concerns with future supply and cost of petroleum-based chemicals as well as with toxic emissions from building materials bonded with some synthetic binders have prompted the wood industry to research renewable materials for wood adhesives. Phenol-formaldehyde (PF) adhesives are used in excess of 2 billion pounds annually in many wood products. Soy protein isolate and soy flour were blended with PF, characterized thermally via differential scanning calorimetry, and analyzed for dry and wet strength along with adhesive failure. At replacement levels <20 wt%, isolate and flour behaved similarly to pure PF, but flour/PF blends showed decreased strength and curing properties ≥20 wt% , whereas isolate/PF blends retained similar strength values to control PF samples. DSC thermal characterization provided a better understanding of adhesive crosslinking during adhesive curing.
AM 3 / PCP 3.1: Plant Protein for Aquafeed
Chair(s): N. Vary, Canadian Food Inspection Agency, Canada; and K. Liu, USDA, ARS, USA
NOAA and USDA's Future of Aquaculture Feeds Initiative. M.B. Rust, Northwest Fisheries Science Center, Seattle, WA, USA
To meet the growing consumer demand for seafood in the United States, increasing supplies of finfish and shellfish will be needed. Most experts agree that development of aquaculture will be the only way to sustainably meet this increase in demand. The question that must be answered is how to ensure that aquaculture production increases are sustainable. The development and expansion of farming of carnivorous fish species will be constrained by a limited supply of fish meal and oil for feeds. Fishmeal and oil have traditionally made up a large part of the diet of farm-raised carnivorous fish. Fortunately, there is no dietary requirement for specific amounts of fish meal or fish oil for fish, so feeds that lessen the reliance on these limited feedstuffs—such as alternative protein and oil resources—can, and must, be developed. For this reason, the U.S. Department of Commerce (NOAA) and the U.S. Department of Agriculture (USDA) is in the process of sponsoring expert and public consultations on the future of fish feeds and the benefits to the U.S. by the development of such alternative feeds.
The Use of Corn Gluten in Salmonid Diets: Issues and Opportunities. G. Vandenberg1, G. Dagenais1, M. de Francisco2, D. Bureau2, 1Université Laval, Quebec, QC, Canada, 2University of Guelph, Guelph, ON, Canada
The main goal of this study was to test the use of white corn gluten in experimental diets to counter the problems associated with the undesirable colouring of trout flesh. The second objective is supplementation of feed with lysine, an essential amino acid naturally found in corn gluten, in order to improve carotenoid binding in fish flesh via protein deposition. A 6-month factorial study (2 x 2; yellow and white corn gluten by presence or absence of supplemental lysine) was undertaken. Rainbow trout (Oncorhynchus mykiss; 250 g initial body mass) were fed with diets that contain 30% of corn gluten and 1.7% of lysine. The colorimetric analyses (CIE L*a*b*) were compared to HPLC xanthophylls analyses. The fish showed a better retention of astaxanthin (p<0,05) when they were fed with lysine-supplemented diet. There are also significant differences in flesh colour according to the type of corn gluten (yellow or white) present in fish feed (p<0,05). A significant effect of the fish weight on the yellow flesh colour (b*) with colorimetric analyses (p<0,001) was shown. The results of this study will enable researchers and aquaculturists to gain a better theoretical understanding of the pigmentation of trout flesh as a result of the effect of lysine, while promoting the use of white corn gluten as a low-polluting, non-colouring, high protein feed ingredient
Selecting Salmonids to Better Utilize Plant Based Diets. K. Overturf, T.G. Gaylord, F. Barrows, USDA, RS, Hagerman Fish Culture Experiment Station, Hagerman, Idaho USA
Evaluation of genotype by diet interactions in aquaculture species for specific dietary components has only recently begun on a limited basis. Initial studies have examined such species as rainbow trout and Atlantic salmon. Because of the high-protein diet these species consume in the wild, commercial diets have relied heavily on fish meal and oil as protein and energy sources. Other omnivorous fish species such as tilapia and catfish have demonstrated a greater proclivity for utilizing plant feedstuffs and carbohydrate for energy, but little research has been performed on these species in regards to precise physiological changes in accord with nutrient modification or selection to alter performance on specific feed types. Research performed in other agriculture animal systems, such as cattle and poultry, have found alterations in specific physiological traits for different strains. These changes have also been found to significantly correlate with individual genes in research done with murine and laboratory fish models. This research is now being applied to aquaculture. Initial findings have varied between stocks and with different diets, but as the technology and experimental designs improve, it appears that this research will prove important for optimizing diets and carnivorous fish species for enhanced utilization of sustainable plant and oil products.
Chemistry and Technology of Processing Canola Proteins for Aquafeed. J. Wanasundara, Agriculture & Agri-Food Canada, Saskatoon, SK, Canada
Processing of canola/rapeseed is primarily to obtain oil. Seed proteins remain in the meal and account up to 42% of the drymatter. Canola meal is a widely used protein source in animal feeds including farmed fish. Processing of meal protein ingredients is not yet an established industry. Biofuel and fuel additive generation from oil increases the domestic canola crushing so does the generated meal volume. A record volume of 3.42 MilT of canola was processed in Canada during 05/06 and is expected to increase in 06/07. Developing high value end uses such as protein ingredients from meal biomass will help the economics of biofuel production and enhance the value of meal components. Canola protein processing includes generation of concentrates and isolates. Composition of proteins and minor components of canola is different than other oilseeds thus approaches to obtain protein products are different. Approaches that are different as well as similar to soy protein processing have been tested for canola. Two of the processes developed in Canada are underway for large scale canola protein ingredient processing. This presentation will provide scientific insight into canola protein processing technologies with reference to the seed chemistry, process technology advances, potential product applications and the significance of protein ingredient development in total seed biomass utilization.
The Utilization of Carotenoids to Improve Fish Health in Aquafeeds. T. Nakano, T. Yamaguchi, M. Sato, Marine Biochemistry Laboratory, Graduate School of Agricultural Science, Tohoku University, Sendai, Miyagi, Japan
Fish oils used in aquaculture is known to contain plentiful highly unsaturated fatty acids (HUFAs). Oxidized HUFA could lead to oxidative stress in fish. Some fish diseases are thought to be catalyzed by oxidative damage to tissue. Hence improvement of the defensive ability of fish against oxidative stress would be important. Carotenoids have been suggested to show many kinds of biological activities in mammals. However, most research on dietary carotenoids, especially astaxanthin (ASX), in aquafeeds has been done in terms of muscle pigmentation, so that information on the biological effects of carotenoids in fish is lacking. Here, we will present several findings of the effect of ASX, including red yeast Phaffia rhodozyma, which is rich in ASX, on the health in rainbow trout#. Diets supplemented with ASX could decrease the levels of lipid peroxides in serum and liver of fish. The elevation of tocopherol concentration in the liver was also observed in fish fed ASX. Furthermore, red yeast is known to contain many kinds of bioactive compounds such as vitamins, trace elements and polysaccharides. Thus we might conclude that ASX and red yeast have many possibilities to improve fish health in aquaculture.#T. Nakano: Dietary Supplements for the Health and Quality of Cultured Fish (Nakagawa, Sato and Gatlin III, Eds), CABI, UK, 86, 2007.
Replacement of Fish Meal and Oil by Canola Protein Concentrate and Vegetable Oils in Diets Fed To Rainbow Trout. M.D. Drew, University of Saskatchewan, Saskatoon, SK, Canada
Replacement of fishmeal and fishoil with plant proteins and oils requires the development of feed ingredients with nutrient profiles similar to marine ingredients. However, such feed ingredients have been difficult to develop. Most plant protein ingredients contain antinutritional factors that limit their use in salmonid diets. An exception to this is canola protein concentrate (CPC). CPC is produced by aqueous extraction of canola meal and has a crude protein content and amino acid profile similar to fishmeal and is also low in antinutritional factors. Rainbow trout fed diets containing CPC had growth rates comparable to those fed diets containing fishmeal. Furthermore, the addition of a 1% of a soluble fraction of CPC to diets fed to rainbow trout significantly increased feed consumption compared to controls. On the oil side, canola and linseed oils have desirable fatty acid composition, due to their high content of n-3 fatty acids and appear to be the best candidates for replacing fishoil in salmonid diets. Recent studies in our laboratory have confirmed that the replacement of fishoil with linseed and canola oils does not diminish growth performance and results in fish fillets with acceptable fatty acid composition, sensory properties and decreased levels of organochlorine contaminants.
PCP 3: Effect of Processing on Protein Functionality
Chair(s): C. Onwulata, USDA, ARS, ERRC, USA; and S. Jung, Iowa State University, USA
Simple Rheology of Mixed Proteins. C. Onwulata, A. Thomas, P. Tomasula, USDA, ARS, ERRC, Wyndmoor, PA, USA
Mixing different proteins to form strong gel networks for food applications may create synergistic increases in viscoelasticity that cannot be achieved with a single protein. In this study, small amplitude oscillatory shear analyses were used to investigate the rheology of calcium caseinate (CC), egg albumin (EA), fish protein isolate (FPI), soy protein isolate (SPI), wheat gluten (WG), and whey protein isolate (WPI) in the presence of wheat flour and glycerol. Blends contained one, two, or three of these proteins. Temperature sweeps revealed that most blends displayed a peak tan δ value (the ratio of viscous modulus to elastic modulus) as the specimen aggregated. Blends containing SPI produced high elastic modulus values and those containing FPI gave low values, indicating strong and weak gels, respectively. CC lowered the temperature at which the elastic modulus and viscous modulus of the blend started to increase sharply. Some blends gave results that indicated synergistic effects. One-protein blends containing EA and WG exhibited their highest tan δ values at 53 and 51°C, respectively, but the blend of the two had its largest tan δ at 38 °C. These results provide evidence that products formulated by blending proteins that act synergistically can be used to create a desired increased viscoelastic effect.
Functionality of Membrane Separated Egg White Proteins. S. Mukhopadhyay, P.M. Tomasula, D. Van Hekken, J.B. Luchansky, R. Kwoczak, Eastern Regional Research Center, USDA, ARS, Wyndmoor, PA, USA
The excellent nutritional and functional properties of liquid egg white (LEW), which is essentially a viscous fat-free protein solution, are exploited in many food preparations. Thermal pasteurization (at 56.6oC for 3.5 min. minimum) is currently used by industry to eliminate the microflora in LEW that limits its shelf life. However, pasteurization only reduces the microflora found in LEW. Crossflow microfiltration has been shown effective in reducing the microflora in milk to limits below that of heat pasteurization. It was the intent of this study to determine if microfiltration of a more viscous food such as LEW was possible. To facilitate MF, LEW was subjected to high shear and diluted (1:2 w/w) with distilled water containing 0.5% NaCl. LEW was then microfiltered using a ceramic membrane with a nominal pore size of 1.4 ïm, at a cross flow velocity of 6 m/s. Permeate flux increased by 148% when pH was adjusted from pH 9 to 6 at 40Â°C. The effect of temperature on permeate flux was inconclusive. SDSâ€“PAGE analysis indicated that the MF process did not alter the protein composition of the permeate relative to the feed LEW. The foaming properties of LEW were retained in the post-filtered samples.
Influence of Pentosans on Functional Properties of a Protein Isolate from Flaxseed (Linum Usitatissimum L.). J.-P. Krause, F. Pudel, PPM Pilot Pflanzenöltechnologie Magdeburg, Germany
Oil from flaxseed or linseed (Linum usitatissimum L.) is an important raw material for food and non-food uses. The protein-rich cake after de-oilification could be serve as a source for highly-functional protein biopolymers. Flaxseeds contain considerable amounts of mucilaginous material which is primarily a mixture of polysaccharides The aim of this study was to prepare pentosan-rich protein fractions from flaxseed meal and to investigate the influence on functional properties. Monolayer behaviour, emulsifying and foaming properties, water and oil binding were studied at the acidic and alkaline maximum of protein solubility to be able to elucidate effects of acidic and neutral pentosans and/or protein-pentosan complexes on functionality.We could reproducibly prepare flaxseed proteins with comparable protein pattern but different amount of pentosans (up to 40 %w/w) by a combined isolation procedure.Distinct differences were found in the pH dependent solubility, water and oil binding capacity.The high pressure transition in monolayer points to the ability in formation of stable layers and could be confirmed with emulsifying and foaming experiments. The isolation procedure used enables the production of protein-pentosan-products with tailored properties.
Extrusion and High-pressure Processing Prior to Aqueous Extraction and Enzyme-Assisted Aqueous Extraction of Full Fat Soybean Flakes. Stephanie Jung, Iowa State University, Center for Crops Utilization Research, USA
Aqueous extraction processing (AEP) of soybean has regained considerable interest during the past decade as an environmentally-friendly alternative to hexane extraction of soybean oil. In this study the effect of extrusion and high-pressure processing (200 and 500 MPa, 25°C, 15 min) applied as a pretreatment for AEP and enzyme-assisted aqueous extraction processing (EAEP; 0.5% Protex 7L) of soybean flakes was compared. The best treatment was EAEP of extruded flakes with oil and protein extraction yields of 90% and 82%, respectively. EAEP also yielded 17% free oil, which was the highest yield achieved. The protein fraction is the major co-product extracted during AEP. The use of an enzyme during the extraction step modified the precipitation of the protein at pH 4.5 depending on the pretreatment applied. The crude protein content and oil content of the isolated soy proteins (ISP) ranged between 65-92% and 10-29%, respectively. Functionality (water and oil holding capacity, apparent viscosity and solubility) of the ISP fractions were compared and related to their SDS PAGE profile, native state, and surface hydrophobicity. This study provides information on the protein fraction recovered during AEP, which will contribute to determining the economical feasibility of this process.
Impact of Membrane Separation, Glycation, and Salt Extraction on the Functional Properties of Soy Protein Isolates. J. Boye, Agriculture & Agri Food Canada, St Hyacinthe, Quebec, Canada
Soybean products are used extensively in food formulation for both their nutritional and functional properties. Processing conditions used in the production of soy protein isolates, concentrates and their fractions often induces changes in their structure which can influence their functional performance in foods. The majority of commercial soy protein products available on the market today are produced from hexane-defatted flakes using iso-electric precipitation followed by spray-drying. To evaluate the impact of process modification on the functionality of soy proteins, a combination of salt extraction and membrane separation techniques at different pHs were used to extract soy proteins from defatted soy flakes and whole soybeans and their functional properties were evaluated. The soy protein isolates produced were further glycated with glucose and carrageenan to determine the impact on functionality. Soy protein isolates obtained generally had protein contents of greater than 90%. Solubility values ranged between 70 – 100% at pH 7.5. pH modifications resulted in significant changes in protein electrophoretic profiles and functionality. Glycation of soy protein isolate and 11S glycinin also modified protein functionality. Significant improvements in functionality were also observed when membrane separation was used. Our results demonstrate that targeted modifications of the conditions used during soy protein processing could offer novel opportunity to develop soy protein isolates with improved functionality.
PCP 4: Advances and Challenges Toward Health Benefits of Proteins
Chair(s): H. Kumagai, Nihon University, Japan; P. Kerr, Solae Co, USA; and H. Ibrahim, Kagoshima University, Japan
Antihypertensive Mechanism of Small Peptides. Toshiro Matsui, Kyushu University, Fukuoka, Japan
The importance of biologically active substances in foods has so far received much attention. There is a growing link between diet and disease prevention, thus the effects of food on disease such as hypertension, diabetes, obesity, osteoporosis and cardiovascular disease are being studied. To date, many functional food products with a health claim can be available in Japan. One of the successful products is an antihypertensive food including angiotensin I-converting enzyme (ACE) inhibitory peptides. However, their underlying mechanism of antihypertensive compounds still remains unclear.To date, a number of active peptides (>400) have already been identified as an in vitro ACE inhibitor. In addition, antihypertensive effect of some functional foods containing such ACE inhibitory peptides has been also demonstrated in mild hypertensive subjects and/or high normal BP subjects. These findings strongly lead us to investigate how active peptide(s) elicits antihypertensive effect in human systemic system. In a series of our studies, we have clarified some physiological functions such as intact absorption into human circulatory blood systems, a transient ACE inhibition of the circulating renin-angiotensin system. Our recent studies also demonstrate that some small peptides can regulate vascular functions via blocking of L-type voltage-gated Ca2+ channel.
Improving the Digestibility of Soybean Proteins by Enzymatic and Chemical Disulfide Bond Cleavage. H. Wang, R. Faris, T. Wang, Iowa State University, Ames, Iowa, USA
Glycinin and soybean trypsin inhibitors are rich in disulfide bonds. Cleavage of these disulfide bonds by reduction is expected to increase the digestibility of soy proteins due to increased susceptibility to protease attacks. Two disulfide cleavage methods, enzymatic reduction and chemical reduction, were applied in this study. The enzymatic treatment was carried out using NADPH-thioredoxin-thioredoxin reductase system (NTS). It was found that nearly all the disulfide bonds were reduced at the treatment conditions. The sulfhydryl content was increased from 8 μmol/g protein of the control to 35 μmol/g protein after NTS reduction. The digestibility of soy protein as measured by pH-Stat method increased by 61 and 29% by trypsin and pancreatin digestion after disulfide bond cleavage by NTS. The digestibility of soy protein showed a highly linear relationship with the free sulfhydryl group content. Chemical cleavage of disulfide bonds was done with metabisulfite. The digestibility of treated soy protein also showed significant improvement, although there was seemingly no significant increase of free sulfhydryl content in the treated samples. The SS reduction has potential to be used in non-heat or low-heat soy protein processing, such as aqueous oil and protein separation.
Development of Oral Tolerance Agent to Reduce Cedar Pollinosis Using Allegen-Polysaccharide Conjugate. Akio Kato1, Akira Saito1, Masakatsu Usi2, 1Department of Biological Chemistry, Yamaguchi University, Yamaguchi, Japan, 2Department of Food Science, National Fisheries University, Yamaguchi, Japan
Since 15-20 percent Japanese are suffering from cedar pollinosis in early spring, the effective therapy is desired to reduce or cure the symptom. Thus, we have developed oral immune tolerance agent for Japanese cedar pollen allergy without any side-effects such as anaphylaxis. The allergenic epitopes of allergen protein Cry j 1 were masked by the Maillard-type conjugation with polysaccharide (GM:galactomannan) to enable the oral administration. Repetitive oral administration of pollen allergen Cry j 1-polysaccharide conjugate induces immune tolerance in allergen-sensitized mice. The oral administration of Cry j 1-GM conjugates induced immune tolerance effectively than unmodified Cry j 1. Immunofluorescence analysis indicated that biotinylated Cry j 1-GM conjugate was efficiently uptaken in gut lumen and phagocytosed by macrophage in gut lumen. The clinical symptom of patients was inhibited by administration of allergen Cry j 1-GM conjugate. The symptom of 10 percents of patients was completely cured and the symptom of 70 percents of patients were greatly suppressed.
The Discovery of Functional Sectrets of Hen's Egg Ovotransferrin: A New Potential for Human Health. H.R. Ibrahim, Dept. Biochemistry & Biotechnology, Faculty of Agriculture, Kagoshima University, Kagoshima, Japan
Eggs are biological reactors that contain all necessary molecules to produce and protect chick life, and thus represents an excellent example of nature's original functional food. Particularly egg albumen is loaded with many bio-active proteins, which have long been thought to play important roles in disease prevention and health promotion. In the post-genomic era, we are facing a big challenge to uncover the biological functions of key proteins of egg albumen and formulate novel candidates for health foods and therapy. Ovotransferrin is a key defense protein in egg albumen and many biological systems. However, its biological function(s) is still poorly understood and remained a mystery for decades. In this review, I will introduce our recent discovery delineating the medically important biological function(s) of OTf. It explores that OTf is a redox-regulated protein with novel SOD-like action, which offer tremendous opportunities for its candidacy in the treatment of oxidative-stress related human diseases with potential use in functional foods.
Challenge to Isolation and Identification of Food-Derived Peptide in Human Blood - Candidate for Real Bioactive Peptide. Kenji Sato, Koji Iwai, Misako Inoue, Noriko Sato, Kyoto Prefectural University, Shimogamo, Kyoto 606-8522, Japan
Enzymatic hydrolysates of food proteins show beneficial activities beyond nutritional value. Then, occurrence of food-derived peptides with biological activities in blood has been speculated. This project aims to prove occurrence of food-derived peptides in blood. Bloods from human volunteer and animal before and after ingestion of food protein hydrolysates were deproteinized with ethanol or TCA. The peptide was captured by SPE using cation exchanger and fractionated by SEC and RP-HPLCs. In some cases, peptides were reacted with PITC before RP-HPLC. The pyroglutamyl peptides were digested with pyroglutamyl aminopeptidase. The liberated pyroGlu was reacted with 2-nitrophenyl hydrazine and the derivative was resolved by RP-HPLC. After oral ingestion of gelatin hydrolysate, peptide form of Hyp in human peripheral blood increased significantly, which indicating occurrence of food-derived peptides. Pro-Hyp and Hyp-Gly were identified as major food-derived collagen peptides. Some food-derived peptides were also detected after ingestion of enzymatic hydrolysates of other food proteins. In addition, occurrence of food-derived pyroglutamyl peptide was detected in the rat portal blood. These food-derived peptides, that resist digestive pepetidase and can be absorbed into blood system, might be good candidate of active peptides.
Production of Rabies Neutralizing Antibody (IgY) in Hen's Eggs using Recombinant Rabies Virus Antigens Expressed in Escherichia Coli. Hajime Hatta1, Yuri Motoi2, Satoshi Inoue2, Kinjiro Morimoto2, 1Kyoto Women's University, Kyoto, Japan, 2National Institute of Infectious Diseases, Tokyo, Japan
In an attempt to produce anti-rabies antibody being available for people living in developing countries, we have immunized hens with recombinant nucleoprotein (rN), phosphoprotein (rP) and a part of surface G protein (GF2) of the rabies virus (CVS-11 strain) expressed in Escherichia coli. Yolk immunoglobulin (IgY) was purified from the yolks of eggs laid by the hyper-immunized hens. The anti-rN and rP IgY were likely to bind specifically to the respective proteins of the CVS-11 strain of rabies virus by Western blotting, immune-fluorescent assay and immunohistochemical detection, indicating these IgY could serve as a reagent for diagnosis of rabies virus infection. On the other hand, only the anti-GF2 IgY apparently neutralized rabies virus infectivity. Inoculation of the anti-GF2 IgY into mice infected with rabies virus reduced the mortality caused by the virus, suggesting that IgY directed to the part of the G protein could serve as a possible alternative to currently available anti-rabies human or equine immunoglobulins.
Proteins Catalyzing the Reaction to Produce Bioactive Compounds: C-S Lyases Activating Sulfuric Compounds to Inhibit Platelet Aggregation. H. Kumagai, Nihon University, 1866 Kameino, Fujisawa-shi 252-8510, Japan
Food processing sometimes improves the functionality of food by producing bioactive compounds enzymatically. During cutting or crushing, isothiocyanates are produced from glucosinolates in Cruciferae plants by myrosinase, whereas sulfides are produced from S-alk(en)yl-L-cysteine sulfoxides in Allium plants by C-S lyase. Some of the isothiocyanates and sulfides have preventive effects on cancer and thrombosis. However, one of the difficulties for using them in food has been their strong flavor, which is not often considered acceptable. If bioactive compounds such as these are produced from their precursors in the body, then the enzymatic reaction during processing would not be necessary and their application would be extended because most precursors are odorless water-soluble compounds. Therefore, we administered S-allyl-L-cysteine sulfoxide (ACSO) to rats to examine if it could be converted into bioactive compounds in vivo. Inhibitory activity against platelet aggregation was observed after the oral administration of ACSO, indicating that ACSO had been converted into sulfides. Then, we sought to identify the protein in the small intestines that was responsible for the catalytic reaction, and found that hemoglobin functioned as a C-S lyase.
Bioactive Peptides from Soy Protein Hydrolysates. Elvira Gonzalez de Mejia, Vermont Dia, Wenyi Wang, University of Illinois, Urbana, Illinois, USA
Food proteins not only provide essential amino acids to meet nutritional requirements, but also exert functional health benefits. Soybean, an important source of food proteins, has received increasing interest from the public because of its reported health benefits. This presentation will provide a comprehensive review of the recently used techniques in the analysis and characterization of soy bioactive peptides including a co-immunoprecipitation approach to identify topoisomerase II inhibitory peptides in soy hydrolysates and their molecular interaction. Soy protein isolate was subjected to simulated gastrointestinal digestion with pepsin and pancreatin. Human topoisomerase II inhibitory peptides in the hydrolysates were co-immunoprecipitated with topoisomerase and identified with CapLC- Micromass Q-TOF Ultima API system. Topoisomerase II inhibition increased during the course of soy hydrolysis from 1% to 61% at 0.5 mg/mL. Three topoisomerase II inhibitory peptides derived from major soy proteins were identified and their IC50 ranged from 2.4 mM to 7.9 mM. The interaction energy with the catalytic domain correlated with the IC50 values (R2 = 0.99). In summary, there are opportunities for industrial exploitation of soy value-added bioactive peptides that can be used to enhance health and prevent disease.
Progress in the Identification of the Cholesterol-Lowering Components in Soy. C. Schasteen, C. Jankovich, J. Wu, B. Pierce, E. Krul, The Solae Company, St. Louis, MO, USA
A key benefit of soy in the diet is its ability to lower blood cholesterol levels. One possible mechanism to lower circulating cholesterol is to bind bile acids in the digestive tract. Cholesterol is the precursor of bile acids, which are used during digestion to emulsify fat and aid in lipid uptake. Normally bile acids are effectively readsorbed back into the body via intestinal transporters, however, not allowing the bile acid to be readsorbed leads to excretion. This bile acid excretion results in a decrease in serum cholesterol levels as the liver extracts cholesterol for bile acid synthesis. We have developed an in vitro screen that quickly measures soy's bile acid binding capacity. This method measures the amount of bile acid which is bound to a sample. The assay has been able to differentiate the binding capacity (μg bile acid per mg test substance) of varied soy protein samples with the finding that W0008, a Solae product which has previously shown to bind bile acids in our screen and lower circulating cholesterol in the male Syrian hamster, an animal model of human lipids.also binds bile acids effectively. This information has been used to screen our products and allow us to isolate the active component in soy and determine a molecular mechanism for soy's ability to reduce cholesterol.
Soy Protein Prevents Hyperinsulinemia and Excess Accumulation of Body Fat in Rats Fed a High Fat Diet. Armando R. Tovar, Nimbe Torres, Depto. Fisiologia de la Nutrición, Instituto Nacional de Ciencias Médicas y Nutrición, Mexico
Obesity is a public health problem that affect many countries around the world. Several consequences of the development of obesity are hypertension, hyperlipidemia, and insulin resistance, leading in the development of cardiovascular diseases and diabetes type 2. During the process of developing overweight and obesity, adipose tissue plays an important role in preventing the deposition of lipids in organs such as liver, skeletal muscle, heart, and pancreas. When adipose tissue is incapable to play this essential role, deposition of lipids occur in these organs leading in fatty liver, or abnormalities in insulin secretion, problems associated with the metabolic syndrome. We have demonstrated in obese rats that consumption of soy protein ameliorates these abnormalities even after the consumption of high fat diets. During the development of obesity there is hyperinsulinemia that is associated to insulin resistance, and consumption of soy protein prevents hyperinsulinemia. This is in part due by stimulating in a lesser extent pancreatic insulin secretion mediated by the amino acid pattern of soy protein, and by isoflavones. In addition, soy protein consumption results in lesser insulin resistance than in rats fed casein determined by hyperglycemic-hyperinsulinemic clamp. These hormonal changes were associated with a low expression of SREBP-1 and high expression of SREBP-2 in liver resulting in low hepatic lipid deposition. On the other hand, long term soy protein consumption prevented hyperleptinemia, and reduced the release of fatty acids from the adipocyte by increasing perilipins mRNA expression despite the high fat content in the diet. In addition, soy protein prevented the increase in the adipocyte area, which was associated with a lower body fat content. Furthermore, the lipid droplet area in brown adipose tissue was significantly lower in rats fed soy protein and it was associated with a higher UCP-1 expression. As a result, rats fed soy protein high fat diet gained less weight in part possibly due to an increase in the thermogenic capacity mediated by UCP-1.
PCP 5: General Topics
Chair(s): P. Qi, USDA, ARS, ERRC, USA; and J. Schmitz, Iowa State University, USA
Extraction of Ovomucin from Egg Whites. J. Wu, A.O. Dileep, University of Alberta, Edmonton, AB, Canada
Hen egg white ovomucin is a highly viscous sulfated glycoprotein, which accounts for approximately 3.5% of egg white proteins. It is characterized by high molecular weight, high carbohydrate content, and a subunit structure (α and β ovomucin). The carbohydrate contents of α and β ovomucin are roughly 15% and 60% respectively. Ovomucin is responsible for the gel like properties of thick egg albumin. Ovomucin usually occur as a complex with lysozyme and other egg white proteins. Previous reported literatures have indicated that ovomucin in hen egg white proteins may have some biological attributes, e.g. antiviral activities, anti-tumor and immunomodulating effects. Although ovomucin can be easily fractionated from egg albumin by dilution of egg albumin with water followed by acidification, further purified by KCl washing, the method of extraction is labor-intensive and consumes a large deal of water for repeated washing. Further, the resulted ovomucin contained many of other egg white proteins. The prepared crude ovomucin is highly insoluble in common buffer solutions, hence usually solubilized in buffers containing dissociating or reducing agents. In the present study, effect of different salt concentration or combinations on ovomucin extraction has been attempted attempting to improve the method of extraction and eliminate the washing step with KCl, if possible.
Exploring the Cross-Linking Reaction Mechanism of Genipin with β-Lactoglobulin and Related Peptides by MALDI-TOF/TOF Mass Spectrometry. P. Qi, A. Nunez, P. Tomasula, US Department of Agriculture, Agricultural Service, Eastern Regional Research Center, Wyndmoor, PA, USA
Genipin has been shown to be capable of cross-linking the primary amine groups of proteins such as the epsilon-amino groups of lysine, and to a lesser extent, of arginine and glutamine. However, the mechanism of the cross-linking reaction is not fully understood. In this study, we examined the reactions of the tryptic peptides of β-lactoglobulin and other synthetic peptides with genipin using MALDI-TOF/TOF mass spectrometry. The results from kinetics studies clearly identified reaction intermediates during the cross-linking reactions and the final products formed subsequently. The reaction intermediates, resulting from nucleophilic attack by the amine at two sites of the genipin molecule, proceeded to successfully cross-link. In addition, a by-product derived from a simple addition of a monomeric genipin molecule to the N-terminus of a peptide, not reported previously, was observed and may cause the termination of the cross-linking reaction as a competitive reaction scheme. The significance of this work lies in establishing appropriate reaction conditions that can be adapted effectively in cross-linking agricultural proteins such as whey proteins using genipin to produce novel food and non-food products.
Thermal and Rheological Properties of Soy Protein-Based Adhesives. Xiaoqun Mo, X. Susan Sun, Kansas State University, Manhattan, KS, USA
Soy protein-based adhesives have shown potential as alternatives to petroleum-based adhesives. The objective of this undertaken is to develop and characterize water-resistant soy-based adhesives. Second-order response surface regression model was used to study the effects of pH, protein concentration, and sodium chloride concentration on water resistance of soy-based adhesives. Results showed that pH, protein concentration, and salt concentration had significant effects on wet adhesion strength. However, no significant effect of sodium chloride concentration on boiling adhesion strength was observed. Thermal stability of soy-based adhesives increased as sodium chloride concentration increased, as indicated by the increased protein denaturation temperatures and denaturation enthalpy. The rheological properties of soy-based adhesives were also found dependent on sodium chloride concentrations. Soy protein adhesive, prepared at pH 5.5 with 28% protein concentration and 1% sodium chloride, had a viscosity of 2.45 Pa s much lower than 18.9 Pa s for the adhesives with no salt added. The adhesive have predicted dry adhesion strength of 6.45 MPa, wet adhesion strength of 3.62 MPa, and boiling adhesion strength of 2.68 MPa.
Protein and Co-Products Posters
Chair(s): M. Dowd, USDA, ARS, SRRC, USA
High Strength Bio-Based Fiberboard Improved by Soy Flour Adhesives.
Xin Li1, Donghai Wang1, Jo A. Ratto2, Xiuzhi Susan Sun1, 1Kansas State University, Manhattan, KS, USA, 2U.S Army Soldier Natick Research, Development and Engineering Center, Natick, MA, USA
Fiberboard manufacture employs formaldehyde based resins currently. However, the formaldehyde resins are harmful to the human health, and mainly made from petroleum products, which reserves are naturally limited. Therefore, the objective of this research was to develop bio-based fiberboard using soybean flour adhesives.The soybean flour adhesives were developed in our lab. Soy flour was modified with 1% of sodium dodecyl sulfate solution first, then the slurry be dried, and then the modified soy flour adhesives were obtained after milling the dried substances. Commercial pulp fiber was used to make fiberboards for this research. Compared to commercial fiberboard V2s for packaging uses, the bio-based fiberboard had stronger strength. This research suggests that the bio-based fiberboard has great potential as alternative to current commercial fiberboard.
Functional Properties of Food Protein and Polysaccharide Mixtures Optimized by Response Surface Methodology.
Arthur Bueno, Cristina Pereira, Inar Castro, Department of Food and Experimental Nutrition, Faculty of Pharmaceutical Sciences, University of São Paulo, Av. Lineu Prestes 580 B14, 05508-900 São Paulo, Brazil
Proteins and polysaccharides can exert a complementary function as emulsifiers in food formulations. Multivariate statistical techniques were first applied to classify different ingredients according to their chemical composition and functional properties. Three ingredients were selected and functional properties of their mixtures were optimized using response surface methodology (RSM). A simplex centroid design was applied to model emulsifying activity index (EAI), average droplet size (D[4,3]), creaming inhibition (CI), emulsion stability (ES) and water retention capacity (WRC) of the mixtures containing different proportions of high methoxyl pectin (x1 ), textured soybean protein (x2 ) and isolated soybean protein (x3 ). All polynomial models showed a good fitness in respect to the experimental data. The proportion of the ingredients suggested by optimization procedure showed an increase of 19% in EAI, 22% in ES, with the same value for D[4,3] (18μm) and WRC (1.4 ml/g) when compared with powdered textured soybean protein. More homogeneous droplet distribution was observed in the optimized mixture than when the ingredients were individually applied. Results can be partially attributed to the electrostatic balance between the anionic polysaccharide and soybean proteins at pH values above the protein isoelectric point.
The Effect of Transglutaminase Crosslinking Reactions on Soy Protein vs. Heated Soy Protein Dispersions.
D.A. Clare1, H. Hwang2, P. Kwanyuen3, C. Daubert1, 1Department of Food, Bioprocessing, and Nutrition Sciences, Raleigh, NC, USA, 2Department of Molecular and Structural Biochemistry, Raleigh, NC, USA, 3USDA-ARS and Crop Science Department, NC State University, Raleigh, NC, USA
Previously, we created a thermally modified soy protein isolate (mSPI) that was readily reconstituted in water and demonstrated improved heat stability and cold-set gel functionality compared to unheated SPI. Herein, we examined the effect of enzyme modification (microbial transglutaminase; mTGase) on equivalently prepared SPI and mSPI protein dispersions with respect to: (1) solubility, (2) the effects of temperature on polymerization reactions, (3) degree of cross-linking, (4) SDS-PAGE protein banding patterns before and after mTGase treatment, and (5) resultant altered functional parameters, such as apparent viscosity, after enzyme processing. Aggregate formation occurred in mSPI solutions which diminished solubility and heat stability. Notably, the mSPI substrate was not cross-linked to the same extent as SPI. Perhaps, this was due in part to aggregate formation that limited accessibility to the TGase catalytic site. SPI and mSPI dispersions exhibited a yield stress and non-Newtonian flow behavior. In contrast, both enzyme treated samples demonstrated shear thinning properties. The apparent viscosity was significantly higher in TGase-treated SPI vs. Tgase treated-mSPI dispersions, both of which were greatly enhanced compared to controls. These differences in functionality will impact the design for future applications.
Use of Sunflower Cake Obtained In Oil Extraction for Adsorbent Production.
Rosana de Cassia de Souza Schneider, Adriane Lawisch Rodriguez, Nelma Spinato Balbinot, Fabrício Ivan Kappel, Marta Barcellos Santos, Universidade de Santa Cruz do Sul, Santa Cruz do Sul, RS, Brazil
The use of co-products of the sunflower biodiesel productive chain has been studied aiming at the production of active charcoal. Charcoal was produced in an oven at 600 and 700 ºC for 20 and 30 minutes, having, as raw material, sunflower cakes of Aguará species, got in laboratory, and at sunflower industry. Activated carbon has been prepared by physical activated methods by pyrolysis at 110 ºC for 24 hours and chemical activation with KOH at 50% (m/v) at 700 ºC for 2 minutes. The activated carbon produced has been assessed by adsorption of blue of methylene in aqueous solution and for iodine adsorption. With chemical activation, all samples have got activity increase, i. e, larger specific surface area. Activated carbons produced are, therefore, comparable to commercial ones. The results of the active charcoal production were compared with results of the direct activation of the sunflower cake and it was observed that the activity was better with active charcoal. Therefore, the production of adsorbents from sunflower cake is viable.
Effect of Environmental Stressors on Biopolymer Particles Formed by Heat Treatment of β-Lactoglobulin/Beet Pectin Complexes.
O.G. Jones, D.J. McClements, University of Massachusetts, Amherst, Amherst, MA, USA
Acid-stable biopolymer complex particles between β-lactoglobulin and beet pectin were prepared using a novel acidification/heating procedure. Incorporation of sodium chloride was tested at different preparative stages to elucidate electrostatic effects. Also, dehydration and freezing were investigated on fully formed particles. After preparation, analyses were performed through visual observation, visible spectrophotometry, dynamic light scattering, and electrophoretic mobility. Salt inclusion prior to full particle formation created unstable particles (d > 1000nm), while later addition led to full stability at all concentrations (d < 350nm). Salts screened electrostatic interactions necessary for complexation, yet caused no reversal after thermal denaturation. In fact, 200mM sodium chloride added after preparation contributed to improved acid stability (pH 2-3). No significant changes were found after freezing and thawing procedures. Dehydration induced occasional precipitation events. Addition of sucrose alleviated aggregation, as did sodium chloride to a lesser extent. Particle structure was postulated as filamentous carbohydrate strands radiating outwards from an amorphous, denatured protein core. Such particles may be ideal for hydrophobic compound entrapment or as fat mimetics.
Promotion of Digestibility and Reduction in Allergenicity of Wheat Gliadin by Deamidation.
Keiko Kato1, Akihiro Suda1, Hitoshi Kumagai2, Hidetoshi Sakurai1, Hitomi Kumagai1, 1Nihon University, 1866 Kameino, Fujisawa-shi 252-8510, Japan, 2Kyoritsu Women's University, 2-2-1 Hitotsubashi, Chiyoda-ku, Tokyo 101-8437, Japan
Deamidation of proteins often changes their properties. We showed that the cation-exchange resin of the carboxylate type was quite effective to deamidate food proteins without causing any detectable hydrolysis of peptide bonds. Phytate-removed deamidated soybean globulin showed enhanced calcium absorption from the intestines, while deamidated wheat gliadin and gluten displayed increased solubility in water and salt solutions, and had an improved foaming property. Wheat gluten has a unique property of extendability, which makes it suitable for use in bakery products. However, the low solubility of gluten in water limits its use in other products and also makes it resistant to digestion. The increased solubility of the deamidated gliadin in water may improve its digestibility because solubilized protein is susceptible to hydrolysis by proteases. Moreover, the deamidated gliadin may have low allergenicity because the epitope structure of gliadin contains some glutamine residues in tandem. In the present study, the digestibility and allergenicity of the deamidated gliadn were examined by using an anti-gliadin antibody, an anti-peptide epitope antibody and sera from patients RAST positive for wheat. Our results indicated that the deamidated gliadin was digested rapidly in the stomach and had low allergenicity.
Properties of Yellow and Oriental Mustard Proteins.
J. Wanasundara, S. Abeysekara, Agriculture & Agri-Food Canada, Saskatoon, SK, Canada
Edible mustard in North America include seeds of Sinapis alba (yellow mustard) and Brassica juncea (oriental and brown mustard) that belong to Cruciferae family. Total protein content of yellow mustard (32%) is higher than brown or oriental mustard (26%). Storage proteins are mainly cruciferins and napins. Two varieties of each mustard type were studied for protein properties. Intact seed proteins have lowest solubility (20 to 30% of total protein) between pH 3 and 4. SDS-PAGE separation of soluble proteins showed primarily napins become soluble at the lowest soluble pHs. Solubility of napin at this pH can be improved by changing ionic strength. Cruciferins and napins of S. alba and B. juncea were isolated and purified. Clear endothermic peak was observed for cruciferin at 87.2±0.3°C for S. alba and 85± 0.5°C for B. juncea at pH 7.0. Napins gave a small endothermic peak which was between 65 and 67°C. Allergenic proteins Sin a 1 and Bra j 1E were present at 0.75% and 0.5% of phosphate buffer (pH 7.4) soluble proteins of S. alba and B. juncea, respectively. Information on mustard protein properties is scarce but essential to devise protein utilization strategies.