Investigation of molecular interactions between β-lactoglobulin and sugar beet pectin by multi-detection HPSEC.
Molecular interactions between β-lactoglobulin (β-LG) and sugar beet pectin (SBP) were studied using online multi-detection high performance size exclusion chromatography (HPSEC) at neutral pH and 50mM ionic strength. The hydrodynamic properties of various interacting polymer fractions were characterized in detail and compared with those of β-LG and SBP. Results showed that ∼6.5% (w/w) of native dimeric β-LG molecules formed complexes with over 35% SBP molecules of varying sizes, 800, 110 and 75 kDa. Although the β-LG molecules bind to SBP molecules of all sizes and shapes, they tend to favor the intermediate (110 kDa) and small sized (75 kDa) SBP molecules. All resulting complexes possess altered shapes and hydrodynamic properties when compared to unbound SBP and β-LG. About half of the interacting β-LG (∼3.5%) molecules were thought to bind to a small amount of non-covalently bound feruloyl groups, possibly present in SBP. When pre-heat treated β-LG and SBP were combined, more than 16% of β-LG formed complexes with at least 45% of SBP molecules of varying sizes, Mw∼750-800, 110, and 55-80 kDa. The complexes formed between β-LG aggregates and/or oligomers and the large SBP molecules (750-800 kDa) adopt the shape of β-LG aggregates, random coil. Both groups of complexes formed between β-LG intermediate (110 kDa) and small sized (55-80 kDa) SBP take on the shape of rigid rod. It was speculated that half of the interacting heat-treated β-LG molecules (∼8%) are complexed with non-covalently bound feruloyl groups in SBP.
Eastern Regional Research Center, Agricultural Research Service, U.S. Department of Agriculture, 600 E. Mermaid Lane, Wyndmoor, PA 19038, USA. Electronic address: Phoebe.Qi@ars.usda.gov.
Phoebe X
PX
Qi
Eastern Regional Research Center, Agricultural Research Service, U.S. Department of Agriculture, 600 E. Mermaid Lane, Wyndmoor, PA 19038, USA.
Hoa K
HK
Chau
Eastern Regional Research Center, Agricultural Research Service, U.S. Department of Agriculture, 600 E. Mermaid Lane, Wyndmoor, PA 19038, USA.
Marshall L
ML
Fishman
Eastern Regional Research Center, Agricultural Research Service, U.S. Department of Agriculture, 600 E. Mermaid Lane, Wyndmoor, PA 19038, USA.
Edward D
ED
Wickham
Eastern Regional Research Center, Agricultural Research Service, U.S. Department of Agriculture, 600 E. Mermaid Lane, Wyndmoor, PA 19038, USA.
Arland T
AT
Hotchkiss
D027461:Beta vulgaris / Q000737:chemistry*; D002850:Chromatography, Gel; D006358:Hot Temperature; D057446:Hydrodynamics; D006863:Hydrogen-Ion Concentration; D007782:Lactoglobulins / Q000737:chemistry / Q000378:metabolism*; D008970:Molecular Weight; D009994:Osmolar Concentration; D010368:Pectins / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding
D007782:Lactoglobulins; D010368:Pectins
Ferulic acid; HPSEC; Hydrodynamic properties; Molecular interactions; Sugar beet pectin; β-Lactoglobulin