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Isolation and Characterization of Protein-Tannin Complexes

Albertz, Megan Lee
http://rave.ohiolink.edu/etdc/view?acc_num=muhonors1209128363

Abstract Details

2008, Bachelor of Science, Miami University, College of Arts and Sciences - Biochemistry.
Tannins have two main properties; they readily bind to protein and they can scavenge radicals for days. When tannin is bound to protein, we hypothesize that the complex will provide persistent antioxidant activity in the gastrointestinal tract, and aid in treatment of inflammatory bowel disease. The interaction between proteins and tannins is inadequately understood, so steps were taken to better understand the interaction. First, a method was developed to analyze the complex, and its formation. Two proteins, bovine serum albumin (BSA) and the C-terminus of the SV-40 peptide were tested for complex formation with the tea polyphenolic epigallocatechin gallate (EGCG), and BSA was selected as the best model. Two reaction conditions, covalent crosslinking with formaldehyde and gentle oxidation at basic pH, were examined and the oxidation condition was selected. Anion exchange HPLC was successful in separating native BSA from the BSA-EGCG complex, but size exclusion HPLC was not successful. Kinetics were run using anion exchange HPLC to determine the rate of formation of the complex. When protein is in excess, the product peak increases in size as a function of time over 24 hours. Work was initiated to determine if protein-tannin complexes formed between tannin and protein in cell culture media or if the complex needed to be formed before it was added to the media. Free EGCG interfered with BSA-EGCG in our analysis methods, so ammonium sulfate precipitation and Sephadex-G10 packed columns were tested as methods for removing the free EGCG from the complex. These steps are preliminary for eventually forming complex and analyzing its antioxidant activity in a cell system.
Ann Hagerman, PhD (Advisor)
Chris Makaroff, PhD (Committee Member)
Neil Danielson, PhD (Committee Member)
35 p.
Biochemistry
tannin; EGCG; polyphenol; protein; antioxidant

Recommended Citations

Citations

  • Albertz, M. L. (2008). Isolation and Characterization of Protein-Tannin Complexes [Undergraduate thesis, Miami University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=muhonors1209128363

    APA Style (7th edition)

  • Albertz, Megan. Isolation and Characterization of Protein-Tannin Complexes. 2008. Miami University, Undergraduate thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=muhonors1209128363.

    MLA Style (8th edition)

  • Albertz, Megan. "Isolation and Characterization of Protein-Tannin Complexes." Undergraduate thesis, Miami University, 2008. http://rave.ohiolink.edu/etdc/view?acc_num=muhonors1209128363

    Chicago Manual of Style (17th edition)

muhonors1209128363
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© 2008, all rights reserved.
This open access ETD is published by Miami University Honors Theses and OhioLINK.