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Reducing Skim Milk Powder Dispersion Turbidity by Dissociation of Casein Micelles at Acidic and Neutral Ph: Physicochemical Properties and Possible Mechanisms

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Reducing Skim Milk Powder Dispersion Turbidity by Dissociation of Casein Micelles at Acidic and Neutral Ph: Physicochemical Properties and Possible Mechanisms University of Tennessee, Knoxville TRACE: Tennessee Research and Creative Exchange Masters Theses Graduate School 5-2019 Reducing skim milk powder dispersion turbidity by dissociation of casein micelles at acidic and neutral pH: Physicochemical properties and possible mechanisms Inseob Choi University of Tennessee, [email protected] Follow this and additional works at: https://trace.tennessee.edu/utk_gradthes Recommended Citation Choi, Inseob, "Reducing skim milk powder dispersion turbidity by dissociation of casein micelles at acidic and neutral pH: Physicochemical properties and possible mechanisms. " Master's Thesis, University of Tennessee, 2019. https://trace.tennessee.edu/utk_gradthes/5407 This Thesis is brought to you for free and open access by the Graduate School at TRACE: Tennessee Research and Creative Exchange. It has been accepted for inclusion in Masters Theses by an authorized administrator of TRACE: Tennessee Research and Creative Exchange. For more information, please contact [email protected]. To the Graduate Council: I am submitting herewith a thesis written by Inseob Choi entitled "Reducing skim milk powder dispersion turbidity by dissociation of casein micelles at acidic and neutral pH: Physicochemical properties and possible mechanisms." I have examined the final electronic copy of this thesis for form and content and recommend that it be accepted in partial fulfillment of the requirements for the degree of Master of Science, with a major in Food Science. Qixin Zhong, Major Professor We have read this thesis and recommend its acceptance: Vermont P. Dia, Tao Wu Accepted for the Council: Dixie L. Thompson Vice Provost and Dean of the Graduate School (Original signatures are on file with official studentecor r ds.) Reducing skim milk powder dispersion turbidity by dissociation of casein micelles at acidic and neutral pH: Physicochemical properties and possible mechanisms A Thesis Presented for the Master of Science Degree The University of Tennessee, Knoxville Inseob Choi May 2019 Copyright © 2018 by Inseob Choi. All rights reserved. ii ACKNOWLEDGEMENTS Dr. Qixin Zhong for accepting me kindly as a master’s student and providing me with research opportunity with great guidance. Dr. Tao Wu and Dr. Vermont Dia for being my committee members. Dr. Philipus Pangloli, Dr. John Dunlap, Dr. Michael Essington, Dr. Edward Wright, and Melanie Stewart for scheduling and assisting experiments. Mary Lamsen, Hao Zhang, Teng Li, Shan Hong, Jennifer Vuia-Riser, Andrea Nieto, Anyi Wang, Nan Li, Dara Smith, and Melody Fagan, Wenfei Xiong, Cuixia Sun, and Tao Wang for being my best friends. My family, Heunggeun Choi, Boklee Kim, and Hyeongseob Choi for being my supporters. USDA National Institute of Food and Agriculture, TEN2015-05921 for supporting the study. iii ABSTRACT Casein micelles comprise 80% of dairy proteins. However, casein micelles as the source for protein fortification in beverages are limited due to its contribution to dispersion turbidity. On the other hand, protein beverages are experiencing substantial market growth and what follows is the demand for new protein ingredients. At acidic conditions, like those commonly found in lots of beverages, dispersions of skim milk powder (SMP) have high turbidity and readily precipitate. In order to circumvent these problems, the dissociation of casein micelles is required to reduce turbidity and improve dispersion stability during storage. Additionally, beverages with neutral pH are preferred to consumers suffering dental problems such as tooth erosion. The objectives of the present study were 1) to reduce turbidity of SMP dispersions and 2) characterize their physicochemical properties after treatment at acidic and neutral pH. To acidify SMP dispersions to pH 2.4-3.0, citric acid or glucono delta-lactone was added, followed by subsequent heating at 60, 70, 80, or 90 °C for 2, 5, 10, 30, or 60 min. A lower pH (i.e., pH 2.4) was more effective than pH 3.0 to reduce the dispersion turbidity (e.g., 223 vs 861 NTU) and the hydrodynamic diameter (e.g., 115 vs 265 nm) after heating at 90 °C for 10 min. At neutral pH, translucent dispersions were obtained with the addition of calcium chelators; sodium tripolyphosphate, trisodium citrate, or sodium hexametaphosphate. Higher concentrations of calcium chelators in SMP dispersions resulted in significant reduction in turbidity values compared to the control (e.g., 230 NTU vs >4000). The amount of dissolved calcium in the SMP serum phase apart from the casein micelles iv increased from ~150 to more than 500 mg/L after treatments at both acidic and neutral pH, indicating the dissociation of casein micelles due to the disruption of colloidal calcium phosphate. The SMP dispersions with modified casein structures can be utilized to produce novel types of beverages. v TABLE OF CONTENTS Chapter 1 Introduction .......................................................................................... 1 1.1 Abstract ....................................................................................................... 2 1.2 Literature review .......................................................................................... 2 1.2.1 Early studies on casein ......................................................................... 2 1.2.2 Protein as a nutrient .............................................................................. 5 1.2.3 Reduction of milk turbidity ..................................................................... 7 1.2.4 Structural changes due to heating of milk .......................................... 11 1.2.5 Effect of acidity on casein-whey protein complexation during heating of milk .............................................................................................................. 13 1.2.6 Significance of hydrophobic interactions on casein micelle structure . 14 1.2.7 Significance of colloidal calcium phosphate (CCP) on casein micelle structure ....................................................................................................... 17 1.2.8 Calcium chelating agents .................................................................... 21 1.3 Hypotheses and objectives........................................................................ 22 References ...................................................................................................... 25 Chapter 2 Reducing turbidity of skim milk powder dispersions by dissolving colloidal calcium phosphate from casein micelles and weakening hydrophobic interaction through acidification and subsequent heating ................................... 41 2.1 Abstract ..................................................................................................... 42 2.2 Materials and methods .............................................................................. 42 2.2.1 Chemicals ........................................................................................... 42 vi 2.2.2 Sample preparation ............................................................................ 43 2.2.3 Turbidity .............................................................................................. 44 2.2.4 Hydrodynamic diameter and particle size distribution ......................... 44 2.2.5 Fluorescence spectrometry ................................................................. 44 2.2.6 Inductively coupled plasma-optical emission spectrometry (ICP-OES) ............................................................................................................................ 45 2.2.7 Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS- PAGE) ........................................................................................................ 46 2.2.8 Determination of free thiol groups (SH) concentration ........................ 46 2.2.9 Scanning transmission electron microscopy (STEM) .......................... 47 2.2.10 Statistical analysis ............................................................................ 48 2.3 Results and discussion .............................................................................. 48 2.3.1 Turbidity, hydrodynamic diameter, and particle size distribution of skim milk powder dispersions .............................................................................. 48 2.3.1.1 Effect of heating duration for samples acidified with citric acid .... 48 2.3.1.2 Effect of heating temperature for samples acidified with citric acid ................................................................................................................. 49 2.3.1.3 Effect of pH for samples acidified with citric acid or GDL ............. 50 2.3.2 Relative role of casein and whey protein on SMP dispersions after acidification and heating ............................................................................. 51 2.3.3 Hydrophobic interactions between proteins in skim milk powder dispersions .................................................................................................. 52 vii 2.3.4 Calcium and phosphorous determination from serum phase of skim milk powder dispersions ............................................................................. 56 2.3.5 Effects of acidification and heating on free thiol groups (SH) concentration and molecular weights of proteins ........................................ 57 2.3.6 Structural change of casein micelles using scanning transmission electron microscopy (STEM) .......................................................................
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