Hetero-Protein Coacervation and Complex Equilibria Between β-lactoglobulin and Lactoferrin

• Main Author: Flanagan, Sean E
• Format: Others
• Published: ScholarWorks@UMass Amherst 2014
• Subjects: Coacervation; Hetero-Protein Coacervation; Protein-Protein Interactions; Complex Equilibrium; Lactoferrin; β-lactoglobulin; Analytical Chemistry; Biochemistry; Food Chemistry; Physical Chemistry; Polymer Chemistry
• Online Access: https://scholarworks.umass.edu/theses/1179; https://scholarworks.umass.edu/cgi/viewcontent.cgi?article=2303&context=theses

Coacervation between the milk proteins β-lactoglobulin (BLG) and Lactoferrin (LF) was studied as a model system for hetero-protein coacervation (HPC). Equilibria among BLG/LF complexes and the corresponding speciation were found to control coacervation, which can be quantitatively monitored by turbidimetry. Several methods were used to assess complexation as a function of LF : BLG (mol/mol) mixing ratio (r). Proton release, calculated from a shift in pH when LF is added to BLG, was used to identify regions of complexation. Dynamic light scattering (DLS) was used to determine regions of complexation by relating complex size to stoichiometry. Isothermal titration calorimetry (ITC) was used to measure enthalpies of binding upon addition of LF to BLG. These results are used to show that coacervation is related to speciation, with the LF(BLG2)2 complex as the coacervating species.