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Po-Wei_MS thesis_final version_0807.pdf (2.11 MB)

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The application of supercritical CO2 technology on modifying beta-lactoglobulin in buttermilk to reduce its antigenicity

Yeh, Po-Wei
http://rave.ohiolink.edu/etdc/view?acc_num=osu1564670043653261

Abstract Details

2019, Master of Science, Ohio State University, Food Science and Technology.
The purpose of this study focuses on modifying β-LG (beta-lactoglobulin) in buttermilk via supercritical CO2 treatment, in order to reduce its antigenicity. In dairy industry, Supercritical Fluid Extraction (SFE) is applied majorly in fat extraction. Yet it is possible to utilize SFE as a reactor in foods. Milk is an essential energy source for infants. However, formula is still needed in some instances and there are some children allergic to dairy based products. Buttermilk powder was used in this study because of its abundant milk fat globule membrane and phospholipids content which has been demonstrated to have positive effect on brain and cognitive development during early infancy, therefore, it is now being considered as a nutritional enhancer in infant formulas. In milk proteins, β-LG is believed to be a prevalent allergen in bovine milk. It was treated in SFE system with food grade CO2 at 100, 150, 200, 250, and 400 bar, with 50 and 75°C in each condition. All analyses were completed in 10% buttermilk suspension (w/v). Buttermilk proteins were examined in SDS-PAGE, Western-blot, and ELISA. Orbitrap Fusion LC-MS/MS and periodic staining were used to check any post-translational modification. Purified β-LG was used to evaluate the cytotoxicity, viability, and inflammatory response in Caco-2 cell line by the mean of LDH, MTT, and IL-8, respectively. The SDS-PAGE quantification showed that the signal intensity of β-LG bands was reduced in a maximum of 50.1% after being processed under 250 bar/75°C for 30 min. Lighter and diffused signals were also found in Western-blot. ELISA tests proved that our treatments can significantly change the β-LG antigenicity in buttermilk (p<0.05). Around 324 Da molecular weight increment was detected on treated β-LG by Orbitrap Fusion. Sugar moieties in glycoproteins were revealed by periodic staining. Whey proteins in treated buttermilk sample showed high viability and 24.9% lower inflammation without any cytotoxicity in Caco-2 cells, compared to the untreated sample. These results showed that antigenicity reduction of β-LG was completed by lactosylation, which has been reported as a possible pathway to reduce the allergenicity in foods. The denaturation of β-LG in supercritical fluid processing is a promising way to solve milk allergy, which is still a problem requiring more attention and further research.
Rafael Jimenez-Flores (Advisor)
MacDonald Wick (Committee Member)
Valente Alvarez (Committee Member)
128 p.
Food Science

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Citations

  • Yeh, P.-W. (2019). The application of supercritical CO2 technology on modifying beta-lactoglobulin in buttermilk to reduce its antigenicity [Master's thesis, Ohio State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=osu1564670043653261

    APA Style (7th edition)

  • Yeh, Po-Wei. The application of supercritical CO2 technology on modifying beta-lactoglobulin in buttermilk to reduce its antigenicity. 2019. Ohio State University, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=osu1564670043653261.

    MLA Style (8th edition)

  • Yeh, Po-Wei. "The application of supercritical CO2 technology on modifying beta-lactoglobulin in buttermilk to reduce its antigenicity." Master's thesis, Ohio State University, 2019. http://rave.ohiolink.edu/etdc/view?acc_num=osu1564670043653261

    Chicago Manual of Style (17th edition)

osu1564670043653261
368
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