Progress 09/01/05 to 08/31/08
Outputs
OUTPUTS: Abstracts Presented at National Meetings: (1)Clare, D.A., Hwang, H.M., Kwanyuen, P., and Daubert, C.R. (2008) The Effect of Transglutaminase Cross-linking Reactions on Soy Protein vs. Heated Soy Protein Dispersions. American Oil Chemist Society (AOCS) - Seattle, Washington; (2)Clare, D.A. and Daubert, C.R. (2008) Transglutaminase Polymerization of a Modified Whey Protein Ingredient. ADSA - Indianapolis, Indiana;(3)Hwang, H.M., Clare, D.A., Kwanyuen, P., and Daubert, C.R. (2008) Effect of Tranglutaminase Polymerization on Biochemical and Functional Properties of Heated and Non-heated Soy Proteins. Ms. Hwang presented this poster at the NCSU Undergraduate Research Symposium, Spring 2008;(4)Lillard, J.S., Clare, D.A., and Daubert, C.R. (2007) Expanding the utility of a modified whey protein ingredient via carbohydrate conjugation. IFT 2007 - Chicago, Illinois;(5)Cramp, G.L., Clare, D.A., Kwanyuen, P., and Daubert, C.R. (2007) Formation and functionality of soy protein-dextran and soy protein-mannose conjugates. IFT 2007 - Chicago, Illinois;(6)Cramp, G. L., Kwanyuen, P., and Daubert, C.R. (2007) Molecular interactions of a modified soy protein isolate. AOCS - Quebec City, Quebec, Canada;(7)Clare, D.A. Lillard, S.J., Ramsey, S.R., Amato, P.M., and C.R. Daubert. (2007) Enhanced functionality of a modified whey protein ingredient upon the addition of calcium. Dairy Ingredients Symposium, International Spray Dried Milk Conference - San Francisco, California;(8)Clare, D.A., Lillard, J., Ramsey, S., Amato, P., and Daubert, C.R. (2007) Calcium effects on the functionality of a modified whey protein ingredient. IFT 2007 - Chicago, Illinois. PARTICIPANTS: *Christopher R. Daubert, Professor; *Debra A. Clare, Senior Researcher; *Prabhasankar Pichan, Postdoctoral Researcher (2005 only); Prachuab Kwanyuen, USDA Research Chemist, Associate Professor, Crop Science, NC State University, Raleigh, NC 27695; *George L. Catignani, Professor; *Grace L. Cramp, Graduate Student; and *John S. Lillard, Graduate Student. The asterisk(*) denotes the Dept. of Food, Bioprocessing, and Nutrition Sciences, NC State University, Raleigh, N.C. 27695-7624. TARGET AUDIENCES: The objectives for this project were designed to modify both whey and soy protein fractions via chemical and enzymatic means, and systematically evaluate altered functional properties. In both protein model systems, possible mechanisms to explain some of these changes was proposed. With accomplishment of these goals, the potential to create new ingredients should be of interest to the whey/soy industries. PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
Whey and soy protein starting materials were chemically (Maillard reaction) and enzymatically (transglutaminase) modified to produce glycoconjugates and protein polymers, respectively. Resultant end products exhibited altered functionality, including apparent viscosity, heat stability, water holding capacity, gelling and emulsion properties. Moreover, scanning electron micrographs revealed differences in the protein structural network. Ultimately, the development of novel food hydrocolloids, serving as highly nutritious food ingredients, will provide new global markets for both the whey and soy industries.
Publications
- Cramp, G.L., Kwanyuen, P., and Daubert, C.R. (2008) Molecular Interactions and Functionality of a Cold-gelling Soy Protein Isolate. J. of Food Science 73: E16-E24.
- Lillard, J.S., Clare, D.A., and Daubert, C.R. (2008) Expanding the Utility of a Modified Whey Protein Ingredient via Carbohydrate Conjugation. J. of Dairy Science - Accepted.
- Clare, D.A., Kwanyuen, P., and Daubert, C.R. (2008) The Effect of Transglutaminase Cross-linking Reactions on the Functionality of Soy Protein vs. Heated Soy Protein Dispersions. In preparation.
- Clare, D.A. and Daubert, C.R. (2008) Transglutaminase Polymerization of a Modified Whey Protein Ingredient. In preparation.
- Clare, D.A., Lillard, S.J., Ramsey, S.R., Amato, P.M., and Daubert, C.R. (2007) Calcium Effects on the Functionality of a Modified Whey Protein Ingredient. J. of Agricultural and Food Chemistry 55: 10932-10940.
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Progress 09/01/06 to 08/31/07
Outputs
I. Glycoprotein formation between a modified whey protein concentrate (mWPC) and soy protein isolate (SPI) with carbohydrate materials: Protein solutions, prepared with mWPC and SPI, were conjugated to dextran under dry heating conditions to form glycoprotein conjugates. Maillard reaction products were formed at pH 3.5 for mWPC-dextran samples compared to pH 7.0 for SPI-dextran conjugates. SDS-PAGE and glycoprotein staining techniques were employed to verify complex formation. OPA analyses showed ~39% coupling using mWPC-dextran complexes. Both preparations exhibited increased viscosity and enhanced emulsion stability compared to control protein solutions. II. Molecular Interactions of a Modified Soy Protein Isolate: SPI was thermally treated at pH 7.0 to yield a mSPI ingredient with expanded functional characteristics. The reaction mechanism is summarized accordingly: (1) A minimum ~8% protein concentration was required for gelation (2) Heat-treated SPI exhibited
higher gelling strength, and (3) hydrogen bonding, hydrophobic interactions, and disulfide bond formation were deemed essential to achieve maximal complex viscosity. Ultimately, thiol groupings appeared to be the most significant factor with respect to defining the "absolute functionality" of reconstituted SPI products. III. Transglutaminase polymerization of mWPC dispersions: Transglutaminase (TGase) polymerization of mWPC was accomplished in the absence of dithiothreitol (DTT) at pH 8.1. SDS-PAGE banding patterns of TGase treated mWPC samples evidenced the formation of high molecular weight polymers. OPA assays showed ~30% crosslinking after a 4h treatment at 37 degrees celsius, while significant precipitation was observed after a 24h treatment period. The apparent viscosity of mWPC dispersions was slightly less after enzymatic modification, and scanning electron micrographs revealed a larger pore-sized diameter in covalently linked adducts. Tgase polymerization of mWPC dispersions
may be used to alter functional parameters. IV. Calcium Effects on the Functionality of a modified Whey Protein Ingredient: Protein solutions, prepared with mWPC, showed a dramatically increased thickening capacity upon the addition of extra calcium ion [mWPC-Ca++]. This formulation exhibited expanded gelation characteristics, especially at 4 degrees celsius. SDS-PAGE revealed various disulfide linked forms of beta-lactoglobulin, BSA, and immunoglobulin under non-reducing electrophoretic conditions. Differential scanning calorimetry showed that mWPC-Ca++ dispersions exhibited a two-fold increase in the amount of bound water compared to control samples. Also, the water holding capacity was enhanced. The structural network of mWPC-Ca++ dispersions revealed large sized spheres upon visualized with SEM, a phenomenon ascribed to an increased surface tension caused by the higher salt content. Ultimately, such attributes may afford distinct advantages for whey-based ingredients intended for
application within food systems.
Impacts
The United States is the largest producer and exporter of whey and soy protein products in the world. Therefore, with expanding growth in the production of whey/soy proteins, there are special needs to improve the functionality of these resources. Presently, many whey/soy byproducts are not considered to be high valued added ingredients, and with the advent of novel glycoprotein additives, such as mWP/mSP-CHO, we anticipate their utilization in the world market serving to replace existing dairy/soy protein based food constituents. Herein, whey and soy protein formulations were modified using both chemical (Maillard) and enzymatic (TGase) means, resulting in an improved functionality. With these studies, we have achieved a better understanding of the molecular mechanisms that are involved, especially with regard to the soy product. Ultimately, such technologies may lead to the design of novel value added protein/glycoprotein ingredients that will benefit the US
agricultural industry.
Publications
- Abstract: Clare, D.A. Lillard, S.J., Ramsey, S.R., Amato, P.M., and C.R. Daubert. (2007): Enhanced functionality of a modified whey protein ingredient upon the addition of calcium. Dairy Ingredients Symposium, International Spray Dried Milk Conference. San Francisco, CA
- Abstract: Clare, D.A., Lillard, J., Ramsey, S., Amato, P., and Daubert, C.R. (2007): Calcium effects on the functionality of a modified whey protein ingredient. IFT 2007, Chicago, Illinois
- Abstract: Lillard, J.S., Clare, D.A., and Daubert, C.R. (2007): Expanding the utility of a modified whey protein ingredient via carbohydrate conjugation. IFT 2007, Chicago, Illinois
- Abstract: Cramp, G.L., Clare, D.A., Kwanyuen, P., and Daubert, C.R. (2007): The formation and functionality of soy protein - dextran and soy protein-mannose conjugates. IFT 2007, Chicago, Illinois
- Abstract: Cramp, G. L., Kwanyuen, P., and Daubert, C.R. (2007): Molecular interactions of a modified soy protein isolate. American Oil Chemists Society (AOCS) Quebec City, Quebec, Canada
- Publication: Clare, D.A., Lillard, J.S., Ramsey, S.R., Amato, P.M., and Daubert, C.R. (2007): Calcium Effects on the Functionality of a Modified Whey Protein Ingredient has been submitted to the Journal of Agriculture and Food Chemistry
- Publication: Cramp, G.L., Kwanyuen, P., and Daubert, C.R. (2007): Molecular Interactions of a Modified Soy Protein Isolate has been submitted to the Journal of Food Science
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Progress 09/01/05 to 09/01/06
Outputs
I. Glycoprotein formation between a modified whey protein concentrate (mWPC) and carbohydrate materials: A modified whey protein concentrate, [US patent #6,261,624 B1], was used to generate glycoconjugates using lactose, dextran or corn syrup. Initially, lactosylation of beta-lactoglobulin occurred during the manufacture of mWPC, generating a glycoprotein ingredient that differed from a commercial WPC (ULTRA 80). In fact, the low shear viscosity of mWPC dispersions was >10x higher than equivalent ULTRA 80 solutions. Additional complex formation between mWPC with either dextran or corn syrup was accomplished under both dry and liquid heating conditions and confirmed through OPA analyses and glycoprotein staining of SDS-PAGE gels. Newly formed glycoproteins were slightly less viscous than control WPC dispersions. Scanning electron micrographs revealed a larger pore diameter in the protein network formed by mWPC-glyco- complexes, suggesting that these samples may exhibit
a higher water holding capacity. II. Formulation of a modified soy protein ingredient: Soy protein isolate was thermally treated at pH 7 according to the previous methodology developed for manufacturing the mWPC ingredient. Accordingly, synthesis of a reconstituted soy powder (mSPI), exhibiting expanded functional characteristics was developed. Initial studies were focused on defining the biochemical mechanisms behind such changes, and to date, these findings may be summarized as follows: (1) A minimum concentration of 8% protein was required for gelation (2) Heat-treated soy isolates exhibited higher gelling strength, and (3) hydrogen bonding, hydrophobic interactions, and disulfide bond formation were deemed essential to achieve maximal complex viscosity. Ultimately, the final configuration of the thiol groupings within the system, appeared to have the most significant impact on the absolute functionality of reconstituted soy protein products. III. Transglutaminase polymerization of
mWPC dispersions: Transglutaminase (TGase) polymerization of mWPC was accomplished in the absence of dithiothreitol, after adjustment of the dispersions to pH 8.1. Apparently, sufficient denaturation of the protein starting material occurred during manufacture of the mWPC ingredient. SDS-PAGE banding patterns, used to monitor the degree of crosslinking, dramatically differed from those observed using ULTRA 80, pH 8.1, prepared at equivalent protein concentrations. After a 24h incubation period, significant precipitation was noted in the modified product reflective of extensive crosslinking. In one example, OPA assays, used to quantitatively evaluate the degree of polymerization, showed 30% crosslinking after a 4h treatment at 37 degrees C. Furthermore, the apparent viscosity of TGase modified mWPC solutions was slightly less than non-treated control samples after a 3h incubation period at 37C. Again, scanning electron micrographs revealed a larger pore-sized diameter, suggesting that
the enzyme mWPC ingredient may exhibit an increased water holding capacity.
Impacts
The United States is the largest producer and exporter of whey and soy protein products in the world. In 2003, 320 million pounds of whey and 65.8 million metric tons of soybeans were manufactured. Presently, only one-half of the total liquid volume of whey, produced during cheese making, is further utilized. The annual yield of whey protein concentrate in the US during 2004 was on the average of 24 million pounds per month produced at an approximate value of $5.00/lb (www. aac.wisc.edu/future). Thus, the total market value for this concentrate whey product was ~$120 million dollars/month. By comparison, the soy industry contributes only 5% of its total production for human nutritional purposes within the US (1), and one half of the total value of the US soybean crop is exported as whole soybeans, soybean meal, and/or soybean oil (www.soystats.com/2004). With expanding growth in the production of whey/soy proteins, there are special needs to improve the functionality
of these resources. Presently, many whey/soy byproducts are not considered to be high valued added ingredients, and with the advent of novel glycoprotein-based protein ingredients, such as mWP/mSP-CHO, we anticipate their utilization in the world market serving to replace existing dairy/soy protein based food constituents. Thus, an understanding of the macromolecular interactions between them is crucial for developing technologies leading to the final formulation of a novel valued-added protein/glycoprotein ingredients that will benefit the US agricultural industry.
Publications
- No publications reported this period
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