Effect of Flow Field (Stirring) on the Heat-Induced Fibrillogenesis of Β-Lactoglobulin in the Presence of Glucose at Neutral pH

Document Type : Research Article

Authors

1 Graduate School of Life Science, Hokkaido University, Sapporo, 060-0810, Hokkaido, JAPAN

2 Faculty of Advanced Life Science, Hokkaido University, Sapporo, 060-0810, Hokkaido, JAPAN

Abstract

The effect of stirring on the amyloid fibrillogenesis of β-Lactoglobulin (βLG) at pH 7 was studied in the presence of glucose (Glu). Fibrillogenesis was carried out by heating the 0.30 mM βLG solution at pH 7 with and without glucose (37.5 mM) for 24 hrs at ≥80C under stirring (250 and 474 rpm) conditions. For control samples, βLG solutions with and without glucose were incubated under unstirred conditions. The flow-induced birefringence method was used to characterize the fibrillogenesis, revealing the coil-stretch transition and pointing out the existence of worm-like flexible fibrils in all samples. Atomic Force Microscopy (AFM) was used as a morphology that clearly showed the flexible fibrils in all samples and also revealed that the fibril lengths shortened on increasing the stirring rate. This shortening in lengths might be possible due to weak hydrophobic interaction at pH 7 resulting in fragmentation of fibrils over stirring. Glucose inhibited the fibrillogenesis
of βLG even on stirring.

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Main Subjects


[1] Flower D.R., North A.C.T., Sansom C.E., The Lipocalin Protein Family-Structure and Sequence Overview, Biochim. Biophys. Acta., 1482: 9-24 (2000).
[2] Hambling S.G., McAlpine A.S., Sawyer L., β-Lactoglobulin, in Advanced Dairy Chemistry-1 Proteins, (P.F. Fox ed.) Elsevier Applied Science, London, pp. 141–190 (1992).
[3] Thompson A., Boland M., Singh H., “Milk Proteins: From Expression to Food”, Academic Press: New York (2008).
[4] Papiz M.Z., Sawyer L., Eliopoulos E.E., North A.C.T., Findlay J.B.C., Sivaprasadarao R., Jones T.A., Newcomer M.E., Kraulis P.J., The Structure of Beta-Lactoglobulin and its Similarity to Plasma Retinol-Binding Protein, Nature, 324: 383-385 (1986).
[5] Uhrinova S., Smith M.H., Jameson G.B., Uhrin D., Sawyer L., Barlow P.N., Structural Changes Accompanying ph-Induced Dissociation of the Beta-Lactoglobulin Dimer, Biochemistry, 39: 3565–3574 (2000).
[6] Aguzzi A., Cell biology: Beyond the Prion Principle, Nature, 459: 924-925 (2009).
[7] Chiti F., Dobson C.M., Protein Misfolding, Functional Amyloid, and Human Disease, Annu. Rev. Biochem., 75: 333-366 (2006).
[8] Gosal W.S., Clark A.H., Ross-Murphy S.B., Fibrillar β-Lactoglobulin Gels:  Part 1. Fibril Formation and Structure, Biomacromolecules, 5: 2408-2419 (2004).
[9] Dumay E.M., Kalichevsky M.T., Cheftel J.C., Characteristics of Pressure-Induced Gels of β-Lactoglobulin at Various Times after Pressure Release, Food Sci. Technol, 31: 10-19 (1998).
[10] Katsuta K., Hatakeyama M., Hiraki J, Isothermal Gelation of Proteins. 1. Urea-Induced Gelation of Whey Proteins and their Gelling Mechanism, Food Hydrocolloids, 11: 367-372 (1997).
[11] Dufour E., Robert P., Renard D., Llamas G., Investigation of β-Lactoglobulin Gelation in Water/Ethanol Solutions, Int. Dairy J., 8: 87-93 (1998).
[12] De Wit J.N., Thermal Behaviour of Bovine β-Lactoglobulin at Temperatures up to 150°C. A Review, Trends Food Sci. Technol. 20: 27−34 (2009).
[13] Krebs M.R.H., Devlin G.L., Donald A.M., Amyloid Fibril-Like Structure Underlies the Aggregate Structure Across the pH Range for β-Lactoglobulin, Biophysical Journal, 96: 5013–5019 (2009).
[14] Loveday S.M., Su J., Rao M.A., Anema S.G., Singh H., Whey protein nanofibrils: Kinetic, Rheological and Morphological Effects of Group IA and IIA Cations, International Dairy Journal, 26: 133-140 (2012).
[15] Liu G., Zhong Q., Dispersible and Thermal Stable Nanofibrils Derived from Glycated Whey Protein, Biomacromolecules, 14: 2146–2153 (2013).
[16] Donald A.M., Aggregation in β-Lactoglobulin, Soft Matter, 4: 1147-1150 (2008).
[17] Heyn T.R., Mayer J., Neumann H.R., Selhuber-Unkel C., Kwade A., Schwarz K., Keppler J.K., The Threshold of Amyloid Aggregation of Beta-Lactoglobulin: Relevant Factor Combinations, Journal of Food Engineering, 283: 110005 (2020).
[18] Akkermans C., Venema P., Rogers S.S., Goot A.J.V.D., Boom R.M., Linden E.V.D., Shear Pulses Nucleate Fibril Aggregation, Food Biophys., 1: 144-150 (2006).
[19] Bolder S.G., Sagis L.M.C., Venema P., Linden E.V.D., Effect of Stirring and Seeding on Whey Protein Fibril Formation., J. Agric. Food Chem., 55: 5661-5669 (2007).
[20] Hill E.K., Krebs B., Goodall D.G., Howlett G.J., Dunstan D.E., Shear Flow Induces Amyloid Fibril Formation, Biomacromolecules, 7: 10-13 (2006).
[21] Sharma R.K., Furusawa K., Fukui A., Sasaki N., Effects of a Flow Field on Amyloid Fibrillogenesis in a β-Lactoglobulin Solution, International Journal of Biological Macromolecules, 70: 490-497 (2014).
[23] Hodge J.E., Dehydrated Foods, Chemistry of Browning Reactions in Model Systems, J. Agric. Food Chem., 1: 928–943 (1953).
[24] Pinto M.D.S., Bouhallab S., Carvalho A.F.D., Henry G., Putaux J.-L., Leonil J., Glucose Slows Down the Heat-Induced Aggregation of β-Lactoglobulin at Neutral pH, J. Agric. Food Chem., 60: 214–219 (2012).
[25] Zhao, D., Zhang X., Xu D., Su G., Li B., Li C., Heat-Induced Amyloid-Like Aggregation of β-Lactoglobulin Affected by Glycation by Α-Dicarbonyl Compounds in a Model Study, Journal of the Science of Food and Agriculture, 100: 607-613 (2021).
[26] Biancalana M., Koide S., Molecular Mechanism of Thioflavin-T Binding to Amyloid Fibrils, Biochim Biophys Acta., 1804: 1405–1412 (2010).
[27] Torza S.J., Shear-Induced Crystallization of Polymers. I. the Four-Roller Apparatus, J. Polym. Sci. Polym. Phys., 13: 43-57 (1975).
[28] Karbasi M., Sánchez-Ferrer A., Adamcik J., Askari G., Madadlou A., Mezzenga R., Covalent β-Lactoglobulin-Maltodextrin Amyloid Fibril Conjugate Prepared by the Maillard Reaction. Food Chemistry, 342: 128388 (2021).