Isolation and Characterization of Protein-Tannin Complexes
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Miami University Honors Theses / OhioLINK
2008
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| Online Access: | http://rave.ohiolink.edu/etdc/view?acc_num=muhonors1209128363 |
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ndltd-OhioLink-oai-etd.ohiolink.edu-muhonors12091283632021-08-03T05:42:16Z Isolation and Characterization of Protein-Tannin Complexes Albertz, Megan Lee Biochemistry tannin EGCG polyphenol protein antioxidant 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. 2008-04-26 English text Miami University Honors Theses / OhioLINK http://rave.ohiolink.edu/etdc/view?acc_num=muhonors1209128363 http://rave.ohiolink.edu/etdc/view?acc_num=muhonors1209128363 unrestricted This thesis or dissertation is protected by copyright: all rights reserved. It may not be copied or redistributed beyond the terms of applicable copyright laws. |
| collection |
NDLTD |
| language |
English |
| sources |
NDLTD |
| topic |
Biochemistry tannin EGCG polyphenol protein antioxidant |
| spellingShingle |
Biochemistry tannin EGCG polyphenol protein antioxidant Albertz, Megan Lee Isolation and Characterization of Protein-Tannin Complexes |
| author |
Albertz, Megan Lee |
| author_facet |
Albertz, Megan Lee |
| author_sort |
Albertz, Megan Lee |
| title |
Isolation and Characterization of Protein-Tannin Complexes |
| title_short |
Isolation and Characterization of Protein-Tannin Complexes |
| title_full |
Isolation and Characterization of Protein-Tannin Complexes |
| title_fullStr |
Isolation and Characterization of Protein-Tannin Complexes |
| title_full_unstemmed |
Isolation and Characterization of Protein-Tannin Complexes |
| title_sort |
isolation and characterization of protein-tannin complexes |
| publisher |
Miami University Honors Theses / OhioLINK |
| publishDate |
2008 |
| url |
http://rave.ohiolink.edu/etdc/view?acc_num=muhonors1209128363 |
| work_keys_str_mv |
AT albertzmeganlee isolationandcharacterizationofproteintannincomplexes |
| _version_ |
1719423832185372672 |