Heat-induced denaturation and aggregation of beta-Lactoglobulin: kinetics of formation of hydrophobic and disulphide-linked aggregates

D Galani, Richard KO Apenten

Research output: Contribution to journalArticle

85 Citations (Scopus)

Abstract

Solutions of a whey protein mixture were subjected to various time/temperature treatments, at pH 6.7. Kinetic and thermodynamic activation parameters for the rates of irreversible denaturation/aggregation of the principal whey protein component-beta-lactoglobulin (beta-1g) were followed by gel permeation. Fast Protein Liquid Chromatography (non-dissociating, non-reducing conditions) and by SDS-PAGE (dissociating, non-reducing conditions). The rate of loss of native beta-1g owing to the formation of disulphide linked protein aggregates (k(SDS-PAGE)) and the rate of formation of aggregates via both covalent and non-covalent bonds (k(GP-FPLC)) showed similar biphasic Arrhenius plots. However, the break of the plot occurred at different points. The k(GP-FPLC) Values were higher than values of k(SDS-PAGE) for all the temperatures examined. There was a similar trend for the thermodynamic activation parameters implying that not all of the beta-1g aggregates through thiol-disulphide interactions. Hydrophobically driven associations occur within the aggregates.
LanguageEnglish
Pages467-476
JournalInternational Journal of Food Science and Technology
Volume34
Issue number5-6
DOIs
Publication statusPublished - Oct 1999

Fingerprint

Lactoglobulins
Denaturation
Disulfides
Agglomeration
Kinetics
Chemical activation
Thermodynamics
Arrhenius plots
Proteins
Liquid chromatography
Sulfhydryl Compounds
Permeation
Gels
Temperature
Hot Temperature

Cite this

@article{99759463293145a091253df6ec977f04,
title = "Heat-induced denaturation and aggregation of beta-Lactoglobulin: kinetics of formation of hydrophobic and disulphide-linked aggregates",
abstract = "Solutions of a whey protein mixture were subjected to various time/temperature treatments, at pH 6.7. Kinetic and thermodynamic activation parameters for the rates of irreversible denaturation/aggregation of the principal whey protein component-beta-lactoglobulin (beta-1g) were followed by gel permeation. Fast Protein Liquid Chromatography (non-dissociating, non-reducing conditions) and by SDS-PAGE (dissociating, non-reducing conditions). The rate of loss of native beta-1g owing to the formation of disulphide linked protein aggregates (k(SDS-PAGE)) and the rate of formation of aggregates via both covalent and non-covalent bonds (k(GP-FPLC)) showed similar biphasic Arrhenius plots. However, the break of the plot occurred at different points. The k(GP-FPLC) Values were higher than values of k(SDS-PAGE) for all the temperatures examined. There was a similar trend for the thermodynamic activation parameters implying that not all of the beta-1g aggregates through thiol-disulphide interactions. Hydrophobically driven associations occur within the aggregates.",
author = "D Galani and Apenten, {Richard KO}",
note = "Reference text: 1. ANEMA SG Reaction kinetics of thermal denaturation of whey proteins in heated reconstituted whole milk JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY 44 : 422 1996 2. APENTEN RKO DETERMINATION OF ENZYME GLOBAL THERMOSTABILITY FROM EQUILIBRIUM AND KINETIC-ANALYSIS OF HEAT INACTIVATION FOOD CHEMISTRY 51 : 15 1994 3. ARMSTRONG J ON FRACTIONATION OF BETA-LACTOGLOBULIN AND ALPHA-LACTALBUMIN BIOCHIMICA ET BIOPHYSICA ACTA 147 : 60 1967 4. BROWNLOW S Bovine beta-lactoglobulin at 1.8 angstrom resolution - Still an enigmatic lipocalin STRUCTURE 5 : 481 1997 5. DANNENBERG F REACTION-KINETICS OF THE DENATURATION OF WHEY PROTEINS IN MILK JOURNAL OF FOOD SCIENCE 53 : 258 1988 6. DANNENBERG F THERMODYNAMIC APPROACH TO KINETICS OF BETA-LACTOGLOBULIN DENATURATION IN HEATED SKIM MILK AND SWEET WHEY MILCHWISSENSCHAFT-MILK SCIENCE INTERNATIONAL 43 : 139 1988 7. GALANI D The comparative heat stability of bovine beta-lactoglobulin in buffer and complex media JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE 74 : 89 1997 8. GEZIMATI J Aggregation and gelation of bovine beta-lactoglobulin, alpha-lactalbumin, and serum albumin MACROMOLECULAR INTERACTIONS IN FOOD TECHNOLOGY 650 : 113 1996 9. GEZIMATI J Heat-induced interactions and gelation of mixtures of bovine beta-lactoglobulin and serum albumin JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY 44 : 804 1996 10. HARWALKAR VR KINETIC-STUDY OF THERMAL-DENATURATION OF PROTEINS IN WHEY MILCHWISSENSCHAFT-MILK SCIENCE INTERNATIONAL 41 : 206 1986 11. HAVEA P Electrophoretic characterization of the protein products formed during heat treatment of whey protein concentrate solutions JOURNAL OF DAIRY RESEARCH 65 : 79 1998 12. HILLIER RM WHEY-PROTEIN DENATURATION IN HEATED MILK AND CHEESE WHEY JOURNAL OF DAIRY RESEARCH 46 : 95 1979 13. HOFFMANN MAM Heat-induced aggregation of beta-lactoglobulin: Role of the free thiol group and disulfide bonds JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY 45 : 2942 1997 14. HUTTON JT THE ORIGIN OF SULFHYDRYL GROUPS IN MILK PROTEINS AND THEIR CONTRIBUTIONS TO COOKED FLAVOR JOURNAL OF DAIRY SCIENCE 35 : 699 1952 15. IAMMETI S J AGR FOOD CHEM 43 : 53 1995 16. IAMMETTI S EUR J BIOCHEM 237 : 106 1996 17. KESSLER HG DEV FOOD PRESERVATIO 5 : 91 1989 18. KONRAD G DTSCH MILCHWIRT 45 : 1134 1994 19. LAEMMLI UK CLEAVAGE OF STRUCTURAL PROTEINS DURING ASSEMBLY OF HEAD OF BACTERIOPHAGE-T4 NATURE 227 : 680 1970 20. LAW AJR HEAT DENATURATION OF BOVINE, CAPRINE AND OVINE WHEY PROTEINS MILCHWISSENSCHAFT-MILK SCIENCE INTERNATIONAL 50 : 384 1995 21. LAW AJR DENATURATION OF THE WHEY PROTEINS IN HEATED MILK AND THEIR INCORPORATION INTO CHEDDAR CHEESE MILCHWISSENSCHAFT-MILK SCIENCE INTERNATIONAL 49 : 63 1994 22. LAW AJR HEAT-INDUCED CHANGES IN THE WHEY PROTEINS AND CASEINS MILCHWISSENSCHAFT-MILK SCIENCE INTERNATIONAL 49 : 125 1994 23. LAW AJR COMPOSITIONAL CHANGES IN CAPRINE WHEY PROTEINS MILCHWISSENSCHAFT-MILK SCIENCE INTERNATIONAL 49 : 674 1994 24. LAW AJR QUANTITATIVE FRACTIONATION OF WHEY PROTEINS BY GEL-PERMEATION FPLC MILCHWISSENSCHAFT-MILK SCIENCE INTERNATIONAL 48 : 663 1993 25. LYSTER RLJ DENATURATION OF ALPHA-LACTALBUMIN AND BETA-LACTOGLOBULIN IN HEATED MILK JOURNAL OF DAIRY RESEARCH 37 : 233 1970 26. MANDERSON GA Effect of heat treatment on the conformation and aggregation of beta-lactoglobulin A, B, and C JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY 46 : 5052 1998 27. MANJI B DETERMINATION OF WHEY-PROTEIN DENATURATION IN HEAT-PROCESSED MILKS - COMPARISON OF 3 METHODS JOURNAL OF DAIRY SCIENCE 70 : 1355 1987 28. MCKENZIE HA MILK PROTEINS CHEM M 2 : 257 1971 29. MCKENZIE HA EFFECT OF PH ON BETA-LACTOGLOBULINS NATURE 214 : 1101 1967 30. MCSWINEY M THERMAL AGGREGATION AND GELATION OF BOVINE BETA-LACTOGLOBULIN FOOD HYDROCOLLOIDS 8 : 441 1994 31. MCSWINEY M THERMAL GELATION AND DENATURATION OF BOVINE BETA-LACTOGLOBULIN-A AND BETA-LACTOGLOBULIN-B JOURNAL OF DAIRY RESEARCH 61 : 221 1994 32. MODLER HW SELECTED PROCESSES TO IMPROVE THE FUNCTIONALITY OF DAIRY INGREDIENTS FOOD TECHNOLOGY 41 : 114 1987 33. MONACO HL CRYSTAL-STRUCTURE OF THE TRIGONAL FORM OF BOVINE BETA-LACTOGLOBULIN AND OF ITS COMPLEX WITH RETINOL AT 2.5-A RESOLUTION JOURNAL OF MOLECULAR BIOLOGY 197 : 695 1987 34. MULVIHILL DM WHEY PROTEINS AND THEIR THERMAL-DENATURATION - A REVIEW IRISH JOURNAL OF FOOD SCIENCE AND TECHNOLOGY 11 : 43 1987 35. OLDFIELD DJ Kinetics of denaturation and aggregation of whey proteins in skim milk heated in an ultra-high temperature (UHT) pilot plant INTERNATIONAL DAIRY JOURNAL 8 : 311 1998 36. OWUSU RK A TEST FOR THE 2-STAGE THERMOINACTIVATION MODEL FOR CHYMOTRYPSIN FOOD CHEMISTRY 48 : 231 1993 37. OWUSU RK HEAT INACTIVATION OF LIPASE FROM PSYCHROTROPHIC PSEUDOMONAS-FLUORESCENS P38 - ACTIVATION PARAMETERS AND ENZYME STABILITY AT LOW OR ULTRA-HIGH TEMPERATURES FOOD CHEMISTRY 44 : 261 1992 38. PAPIZ MZ THE STRUCTURE OF BETA-LACTOGLOBULIN AND ITS SIMILARITY TO PLASMA RETINOL-BINDING PROTEIN NATURE 324 : 383 1986 39. ROEFS SPFM A MODEL FOR THE DENATURATION AND AGGREGATION OF BETA-LACTOGLOBULIN EUROPEAN JOURNAL OF BIOCHEMISTRY 226 : 883 1994 40. SAWYER L STRUCTURE AND FUNCTION OF BOVINE BETA-LACTOGLOBULIN BIOCHEMICAL SOCIETY TRANSACTIONS 13 : 265 1985 41. SAWYER WH THERMODENATURATION OF BOVINE BETA-LACTOGLOBULIN KINETICS AND INTRODUCTION OF BETA-STRUCTURE BIOCHIMICA ET BIOPHYSICA ACTA 243 : 19 1971 42. SAWYER WH HEAT DENATURATION OF BOVINE BETA-LACTOGLOBULINS AND RELEVANCE OF DISULFIDE AGGREGATION JOURNAL OF DAIRY SCIENCE 51 : 323 1968 43. VERHEUL M Kinetics of heat-induced aggregation of beta-lactoglobulin JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY 46 : 896 1998",
year = "1999",
month = "10",
doi = "10.1046/j.1365-2621.1999.00314.x",
language = "English",
volume = "34",
pages = "467--476",
journal = "International Journal of Food Science and Technology",
issn = "0950-5423",
number = "5-6",

}

Heat-induced denaturation and aggregation of beta-Lactoglobulin: kinetics of formation of hydrophobic and disulphide-linked aggregates. / Galani, D; Apenten, Richard KO.

In: International Journal of Food Science and Technology, Vol. 34, No. 5-6, 10.1999, p. 467-476.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Heat-induced denaturation and aggregation of beta-Lactoglobulin: kinetics of formation of hydrophobic and disulphide-linked aggregates

AU - Galani, D

AU - Apenten, Richard KO

N1 - Reference text: 1. ANEMA SG Reaction kinetics of thermal denaturation of whey proteins in heated reconstituted whole milk JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY 44 : 422 1996 2. APENTEN RKO DETERMINATION OF ENZYME GLOBAL THERMOSTABILITY FROM EQUILIBRIUM AND KINETIC-ANALYSIS OF HEAT INACTIVATION FOOD CHEMISTRY 51 : 15 1994 3. ARMSTRONG J ON FRACTIONATION OF BETA-LACTOGLOBULIN AND ALPHA-LACTALBUMIN BIOCHIMICA ET BIOPHYSICA ACTA 147 : 60 1967 4. BROWNLOW S Bovine beta-lactoglobulin at 1.8 angstrom resolution - Still an enigmatic lipocalin STRUCTURE 5 : 481 1997 5. DANNENBERG F REACTION-KINETICS OF THE DENATURATION OF WHEY PROTEINS IN MILK JOURNAL OF FOOD SCIENCE 53 : 258 1988 6. DANNENBERG F THERMODYNAMIC APPROACH TO KINETICS OF BETA-LACTOGLOBULIN DENATURATION IN HEATED SKIM MILK AND SWEET WHEY MILCHWISSENSCHAFT-MILK SCIENCE INTERNATIONAL 43 : 139 1988 7. GALANI D The comparative heat stability of bovine beta-lactoglobulin in buffer and complex media JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE 74 : 89 1997 8. GEZIMATI J Aggregation and gelation of bovine beta-lactoglobulin, alpha-lactalbumin, and serum albumin MACROMOLECULAR INTERACTIONS IN FOOD TECHNOLOGY 650 : 113 1996 9. GEZIMATI J Heat-induced interactions and gelation of mixtures of bovine beta-lactoglobulin and serum albumin JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY 44 : 804 1996 10. HARWALKAR VR KINETIC-STUDY OF THERMAL-DENATURATION OF PROTEINS IN WHEY MILCHWISSENSCHAFT-MILK SCIENCE INTERNATIONAL 41 : 206 1986 11. HAVEA P Electrophoretic characterization of the protein products formed during heat treatment of whey protein concentrate solutions JOURNAL OF DAIRY RESEARCH 65 : 79 1998 12. HILLIER RM WHEY-PROTEIN DENATURATION IN HEATED MILK AND CHEESE WHEY JOURNAL OF DAIRY RESEARCH 46 : 95 1979 13. HOFFMANN MAM Heat-induced aggregation of beta-lactoglobulin: Role of the free thiol group and disulfide bonds JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY 45 : 2942 1997 14. HUTTON JT THE ORIGIN OF SULFHYDRYL GROUPS IN MILK PROTEINS AND THEIR CONTRIBUTIONS TO COOKED FLAVOR JOURNAL OF DAIRY SCIENCE 35 : 699 1952 15. IAMMETI S J AGR FOOD CHEM 43 : 53 1995 16. IAMMETTI S EUR J BIOCHEM 237 : 106 1996 17. KESSLER HG DEV FOOD PRESERVATIO 5 : 91 1989 18. KONRAD G DTSCH MILCHWIRT 45 : 1134 1994 19. LAEMMLI UK CLEAVAGE OF STRUCTURAL PROTEINS DURING ASSEMBLY OF HEAD OF BACTERIOPHAGE-T4 NATURE 227 : 680 1970 20. LAW AJR HEAT DENATURATION OF BOVINE, CAPRINE AND OVINE WHEY PROTEINS MILCHWISSENSCHAFT-MILK SCIENCE INTERNATIONAL 50 : 384 1995 21. LAW AJR DENATURATION OF THE WHEY PROTEINS IN HEATED MILK AND THEIR INCORPORATION INTO CHEDDAR CHEESE MILCHWISSENSCHAFT-MILK SCIENCE INTERNATIONAL 49 : 63 1994 22. LAW AJR HEAT-INDUCED CHANGES IN THE WHEY PROTEINS AND CASEINS MILCHWISSENSCHAFT-MILK SCIENCE INTERNATIONAL 49 : 125 1994 23. LAW AJR COMPOSITIONAL CHANGES IN CAPRINE WHEY PROTEINS MILCHWISSENSCHAFT-MILK SCIENCE INTERNATIONAL 49 : 674 1994 24. LAW AJR QUANTITATIVE FRACTIONATION OF WHEY PROTEINS BY GEL-PERMEATION FPLC MILCHWISSENSCHAFT-MILK SCIENCE INTERNATIONAL 48 : 663 1993 25. LYSTER RLJ DENATURATION OF ALPHA-LACTALBUMIN AND BETA-LACTOGLOBULIN IN HEATED MILK JOURNAL OF DAIRY RESEARCH 37 : 233 1970 26. MANDERSON GA Effect of heat treatment on the conformation and aggregation of beta-lactoglobulin A, B, and C JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY 46 : 5052 1998 27. MANJI B DETERMINATION OF WHEY-PROTEIN DENATURATION IN HEAT-PROCESSED MILKS - COMPARISON OF 3 METHODS JOURNAL OF DAIRY SCIENCE 70 : 1355 1987 28. MCKENZIE HA MILK PROTEINS CHEM M 2 : 257 1971 29. MCKENZIE HA EFFECT OF PH ON BETA-LACTOGLOBULINS NATURE 214 : 1101 1967 30. MCSWINEY M THERMAL AGGREGATION AND GELATION OF BOVINE BETA-LACTOGLOBULIN FOOD HYDROCOLLOIDS 8 : 441 1994 31. MCSWINEY M THERMAL GELATION AND DENATURATION OF BOVINE BETA-LACTOGLOBULIN-A AND BETA-LACTOGLOBULIN-B JOURNAL OF DAIRY RESEARCH 61 : 221 1994 32. MODLER HW SELECTED PROCESSES TO IMPROVE THE FUNCTIONALITY OF DAIRY INGREDIENTS FOOD TECHNOLOGY 41 : 114 1987 33. MONACO HL CRYSTAL-STRUCTURE OF THE TRIGONAL FORM OF BOVINE BETA-LACTOGLOBULIN AND OF ITS COMPLEX WITH RETINOL AT 2.5-A RESOLUTION JOURNAL OF MOLECULAR BIOLOGY 197 : 695 1987 34. MULVIHILL DM WHEY PROTEINS AND THEIR THERMAL-DENATURATION - A REVIEW IRISH JOURNAL OF FOOD SCIENCE AND TECHNOLOGY 11 : 43 1987 35. OLDFIELD DJ Kinetics of denaturation and aggregation of whey proteins in skim milk heated in an ultra-high temperature (UHT) pilot plant INTERNATIONAL DAIRY JOURNAL 8 : 311 1998 36. OWUSU RK A TEST FOR THE 2-STAGE THERMOINACTIVATION MODEL FOR CHYMOTRYPSIN FOOD CHEMISTRY 48 : 231 1993 37. OWUSU RK HEAT INACTIVATION OF LIPASE FROM PSYCHROTROPHIC PSEUDOMONAS-FLUORESCENS P38 - ACTIVATION PARAMETERS AND ENZYME STABILITY AT LOW OR ULTRA-HIGH TEMPERATURES FOOD CHEMISTRY 44 : 261 1992 38. PAPIZ MZ THE STRUCTURE OF BETA-LACTOGLOBULIN AND ITS SIMILARITY TO PLASMA RETINOL-BINDING PROTEIN NATURE 324 : 383 1986 39. ROEFS SPFM A MODEL FOR THE DENATURATION AND AGGREGATION OF BETA-LACTOGLOBULIN EUROPEAN JOURNAL OF BIOCHEMISTRY 226 : 883 1994 40. SAWYER L STRUCTURE AND FUNCTION OF BOVINE BETA-LACTOGLOBULIN BIOCHEMICAL SOCIETY TRANSACTIONS 13 : 265 1985 41. SAWYER WH THERMODENATURATION OF BOVINE BETA-LACTOGLOBULIN KINETICS AND INTRODUCTION OF BETA-STRUCTURE BIOCHIMICA ET BIOPHYSICA ACTA 243 : 19 1971 42. SAWYER WH HEAT DENATURATION OF BOVINE BETA-LACTOGLOBULINS AND RELEVANCE OF DISULFIDE AGGREGATION JOURNAL OF DAIRY SCIENCE 51 : 323 1968 43. VERHEUL M Kinetics of heat-induced aggregation of beta-lactoglobulin JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY 46 : 896 1998

PY - 1999/10

Y1 - 1999/10

N2 - Solutions of a whey protein mixture were subjected to various time/temperature treatments, at pH 6.7. Kinetic and thermodynamic activation parameters for the rates of irreversible denaturation/aggregation of the principal whey protein component-beta-lactoglobulin (beta-1g) were followed by gel permeation. Fast Protein Liquid Chromatography (non-dissociating, non-reducing conditions) and by SDS-PAGE (dissociating, non-reducing conditions). The rate of loss of native beta-1g owing to the formation of disulphide linked protein aggregates (k(SDS-PAGE)) and the rate of formation of aggregates via both covalent and non-covalent bonds (k(GP-FPLC)) showed similar biphasic Arrhenius plots. However, the break of the plot occurred at different points. The k(GP-FPLC) Values were higher than values of k(SDS-PAGE) for all the temperatures examined. There was a similar trend for the thermodynamic activation parameters implying that not all of the beta-1g aggregates through thiol-disulphide interactions. Hydrophobically driven associations occur within the aggregates.

AB - Solutions of a whey protein mixture were subjected to various time/temperature treatments, at pH 6.7. Kinetic and thermodynamic activation parameters for the rates of irreversible denaturation/aggregation of the principal whey protein component-beta-lactoglobulin (beta-1g) were followed by gel permeation. Fast Protein Liquid Chromatography (non-dissociating, non-reducing conditions) and by SDS-PAGE (dissociating, non-reducing conditions). The rate of loss of native beta-1g owing to the formation of disulphide linked protein aggregates (k(SDS-PAGE)) and the rate of formation of aggregates via both covalent and non-covalent bonds (k(GP-FPLC)) showed similar biphasic Arrhenius plots. However, the break of the plot occurred at different points. The k(GP-FPLC) Values were higher than values of k(SDS-PAGE) for all the temperatures examined. There was a similar trend for the thermodynamic activation parameters implying that not all of the beta-1g aggregates through thiol-disulphide interactions. Hydrophobically driven associations occur within the aggregates.

U2 - 10.1046/j.1365-2621.1999.00314.x

DO - 10.1046/j.1365-2621.1999.00314.x

M3 - Article

VL - 34

SP - 467

EP - 476

JO - International Journal of Food Science and Technology

T2 - International Journal of Food Science and Technology

JF - International Journal of Food Science and Technology

SN - 0950-5423

IS - 5-6

ER -