Evaluation of a sulphydryl-disulphide exchange index (SEI) for whey proteins - beta-lactoglobulin and bovine serum albumin

Richard Owusu-Apenten, C Chee, OP Hwee

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Sulphydryl-disulphide (SH-SS) exchange underlies many protein functions in processed foods. There is a need for reliable indicators of SH-SS exchange capacity in protein ingredients. An index for SH-SS exchange (SEI) is described using beta-lactoglobulin (BLG) or bovine serum albumin (BSA). The proteins were reacted with 2-pyridine disulphide (PDS Aldrithiol-2(TM)) or Ellman's reagent (DTNB) in a buffered medium (0.05 M phosphate buffer, pH 6.9) at 25degreesC. The 2nd order rate constant for protein SH-SS exchange with PDS or DTNB (k, M-1 s(-1)) was normalized by dividing by the rate constant for glutathione (GSH) reaction with PDS or DTNB (k*, M-1 s(-1)) determined under identical conditions. The capacity or SH-SS exchange was inversely related to SEI (= -log k/k*) values of 4.11 for BLG and 1.05 for BSA, based on measurements using PDS. Studies using DTNB yield SEI equal to 4.28 for BLG and 2.20 for BSA. The SH group of BSA was 100-1052 times more reactive than the SH group of BLG. In a medium containing 1.2 M sodium chloride the difference in SH-SS exchange capacity was similar to9000 fold. Differential scanning calorimetry (DSC) results showed that the conformational stability of BLG increased much more substantially than BSA when both proteins were exposed to 0.1-1.2 M sodium chloride concentrations. (C) 2003 Elsevier Ltd. All rights reserved.
LanguageEnglish
Pages541-545
JournalFood Chemistry
Volume83
Issue number4
DOIs
Publication statusPublished - Dec 2003

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Dithionitrobenzoic Acid
Lactoglobulins
beta-lactoglobulin
bovine serum albumin
Bovine Serum Albumin
whey protein
sulfides
Disulfides
Proteins
proteins
Sodium Chloride
sodium chloride
Rate constants
Processed foods
pyridines
Differential Scanning Calorimetry
processed foods
differential scanning calorimetry
Glutathione
glutathione

Keywords

  • STABILITY FUNCTION RELATIONS
  • CONFORMATIONAL STABILITY
  • THIOL-GROUPS
  • GELATION
  • SALTS

Cite this

Owusu-Apenten, Richard ; Chee, C ; Hwee, OP. / Evaluation of a sulphydryl-disulphide exchange index (SEI) for whey proteins - beta-lactoglobulin and bovine serum albumin. 2003 ; Vol. 83, No. 4. pp. 541-545.
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abstract = "Sulphydryl-disulphide (SH-SS) exchange underlies many protein functions in processed foods. There is a need for reliable indicators of SH-SS exchange capacity in protein ingredients. An index for SH-SS exchange (SEI) is described using beta-lactoglobulin (BLG) or bovine serum albumin (BSA). The proteins were reacted with 2-pyridine disulphide (PDS Aldrithiol-2(TM)) or Ellman's reagent (DTNB) in a buffered medium (0.05 M phosphate buffer, pH 6.9) at 25degreesC. The 2nd order rate constant for protein SH-SS exchange with PDS or DTNB (k, M-1 s(-1)) was normalized by dividing by the rate constant for glutathione (GSH) reaction with PDS or DTNB (k*, M-1 s(-1)) determined under identical conditions. The capacity or SH-SS exchange was inversely related to SEI (= -log k/k*) values of 4.11 for BLG and 1.05 for BSA, based on measurements using PDS. Studies using DTNB yield SEI equal to 4.28 for BLG and 2.20 for BSA. The SH group of BSA was 100-1052 times more reactive than the SH group of BLG. In a medium containing 1.2 M sodium chloride the difference in SH-SS exchange capacity was similar to9000 fold. Differential scanning calorimetry (DSC) results showed that the conformational stability of BLG increased much more substantially than BSA when both proteins were exposed to 0.1-1.2 M sodium chloride concentrations. (C) 2003 Elsevier Ltd. All rights reserved.",
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note = "Reference text: 1. ANDREWS AT 20 INT DAIR C E 1978 162 2. APENTEN RKO Protein stability function relations: beta-lactoglobulin-A sulphydryl group reactivity and its relationship to protein unfolding stability INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES 23 : 19 1998 3. APENTEN RKO Protein stability function relations: native beta-lactoglobulin sulphhydryl-disulphide exchange with PDS JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE 80 : 447 2000 4. BATRA PP CIRCULAR DICHROIC STUDY OF THE CONFORMATIONAL STABILITY OF SULFHYDRYL-BLOCKED BOVINE SERUM-ALBUMIN INTERNATIONAL JOURNAL OF BIOCHEMISTRY 21 : 857 1989 5. BROWNE KR J CONT LEGAL ISSUES 8 : 5 1997 6. CALVO MM INT DAIRY J 3 : 719 1993 7. CASPER JL Seasonal changes in protein composition of whey from commercial manufacture of caprine and ovine specialty cheeses JOURNAL OF DAIRY SCIENCE 81 : 3117 1998 8. FRIEDMAN M CHEM BIOCH SULFHYDRY : 1973 9. HILL AR THE BETA-LACTOGLOBULIN-KAPPA-CASEIN COMPLEX CANADIAN INSTITUTE OF FOOD SCIENCE AND TECHNOLOGY JOURNAL-JOURNAL DE L INSTITUT CANADIEN DE SCIENCE ET TECHNOLOGIE ALIMENTAIRES 22 : 120 1989 10. HILLIER RM GELATION OF RECONSTITUTED WHEY POWDERS BY HEAT JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE 31 : 1152 1980 11. HOWELL NK INT J FOOD SCI TECH 30 : 311 1995 12. KELLA NKD STRUCTURAL STABILITY OF BETA-LACTOGLOBULIN IN THE PRESENCE OF KOSMOTROPIC SALTS - A KINETIC AND THERMODYNAMIC STUDY INTERNATIONAL JOURNAL OF PEPTIDE AND PROTEIN RESEARCH 32 : 396 1988 13. LEE JY EFFECT OF SALTS ON THE THIOL-DEPENDENT GELATION OF BOVINE SERUM-ALBUMIN AGRICULTURAL AND BIOLOGICAL CHEMISTRY 55 : 2057 1991 14. MANNING PB METHOD FOR DETERMINING SULFHYDRYL AND DISULFIDE GROUPS IN MILK PROTEINS JOURNAL OF DAIRY SCIENCE 52 : 886 1969 15. MATSUDOMI N GELATION OF BOVINE SERUM-ALBUMIN AND BETA-LACTOGLOBULIN - EFFECTS OF PH, SALTS AND THIOL REAGENTS FOOD CHEMISTRY 40 : 55 1991 16. OWUSUAPENTEN RK Revised conformational stability parameters for beta-lactoglobulin isothermal denaturation by urea JOURNAL OF FOOD BIOCHEMISTRY 26 : 181 2002 17. RALSTON GB DECREASE IN STABILITY OF BETA-LACTOGLOBULIN ON BLOCKING SULFHYDRYL GROUP COMPTES RENDUS DES TRAVAUX DU LABORATOIRE CARLSBERG 38 : 499 1972 18. REINER CK ANAL BIOANAL CHEM 373 : 266 2002 19. SAKAI K Conformation and stability of thiol-modified bovine beta-lactoglobulin PROTEIN SCIENCE 9 : 1719 2000 20. WALSTRA P DAIRY TECHNOLOGY : 1990 21. WILSON JM SPECTROPHOTOMETRIC METHOD FOR STUDYING THE RATES OF REACTION OF DISULFIDES WITH PROTEIN THIOL-GROUPS APPLIED TO BOVINE SERUM-ALBUMIN JOURNAL OF THE AMERICAN CHEMICAL SOCIETY 102 : 359 1980 22. ZHANG H A kinetic approach to characterize the electrostatic environments of thiol groups in proteins BIOORGANIC CHEMISTRY 26 : 356 1998",
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Evaluation of a sulphydryl-disulphide exchange index (SEI) for whey proteins - beta-lactoglobulin and bovine serum albumin. / Owusu-Apenten, Richard; Chee, C; Hwee, OP.

Vol. 83, No. 4, 12.2003, p. 541-545.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Evaluation of a sulphydryl-disulphide exchange index (SEI) for whey proteins - beta-lactoglobulin and bovine serum albumin

AU - Owusu-Apenten, Richard

AU - Chee, C

AU - Hwee, OP

N1 - Reference text: 1. ANDREWS AT 20 INT DAIR C E 1978 162 2. APENTEN RKO Protein stability function relations: beta-lactoglobulin-A sulphydryl group reactivity and its relationship to protein unfolding stability INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES 23 : 19 1998 3. APENTEN RKO Protein stability function relations: native beta-lactoglobulin sulphhydryl-disulphide exchange with PDS JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE 80 : 447 2000 4. BATRA PP CIRCULAR DICHROIC STUDY OF THE CONFORMATIONAL STABILITY OF SULFHYDRYL-BLOCKED BOVINE SERUM-ALBUMIN INTERNATIONAL JOURNAL OF BIOCHEMISTRY 21 : 857 1989 5. BROWNE KR J CONT LEGAL ISSUES 8 : 5 1997 6. CALVO MM INT DAIRY J 3 : 719 1993 7. CASPER JL Seasonal changes in protein composition of whey from commercial manufacture of caprine and ovine specialty cheeses JOURNAL OF DAIRY SCIENCE 81 : 3117 1998 8. FRIEDMAN M CHEM BIOCH SULFHYDRY : 1973 9. HILL AR THE BETA-LACTOGLOBULIN-KAPPA-CASEIN COMPLEX CANADIAN INSTITUTE OF FOOD SCIENCE AND TECHNOLOGY JOURNAL-JOURNAL DE L INSTITUT CANADIEN DE SCIENCE ET TECHNOLOGIE ALIMENTAIRES 22 : 120 1989 10. HILLIER RM GELATION OF RECONSTITUTED WHEY POWDERS BY HEAT JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE 31 : 1152 1980 11. HOWELL NK INT J FOOD SCI TECH 30 : 311 1995 12. KELLA NKD STRUCTURAL STABILITY OF BETA-LACTOGLOBULIN IN THE PRESENCE OF KOSMOTROPIC SALTS - A KINETIC AND THERMODYNAMIC STUDY INTERNATIONAL JOURNAL OF PEPTIDE AND PROTEIN RESEARCH 32 : 396 1988 13. LEE JY EFFECT OF SALTS ON THE THIOL-DEPENDENT GELATION OF BOVINE SERUM-ALBUMIN AGRICULTURAL AND BIOLOGICAL CHEMISTRY 55 : 2057 1991 14. MANNING PB METHOD FOR DETERMINING SULFHYDRYL AND DISULFIDE GROUPS IN MILK PROTEINS JOURNAL OF DAIRY SCIENCE 52 : 886 1969 15. MATSUDOMI N GELATION OF BOVINE SERUM-ALBUMIN AND BETA-LACTOGLOBULIN - EFFECTS OF PH, SALTS AND THIOL REAGENTS FOOD CHEMISTRY 40 : 55 1991 16. OWUSUAPENTEN RK Revised conformational stability parameters for beta-lactoglobulin isothermal denaturation by urea JOURNAL OF FOOD BIOCHEMISTRY 26 : 181 2002 17. RALSTON GB DECREASE IN STABILITY OF BETA-LACTOGLOBULIN ON BLOCKING SULFHYDRYL GROUP COMPTES RENDUS DES TRAVAUX DU LABORATOIRE CARLSBERG 38 : 499 1972 18. REINER CK ANAL BIOANAL CHEM 373 : 266 2002 19. SAKAI K Conformation and stability of thiol-modified bovine beta-lactoglobulin PROTEIN SCIENCE 9 : 1719 2000 20. WALSTRA P DAIRY TECHNOLOGY : 1990 21. WILSON JM SPECTROPHOTOMETRIC METHOD FOR STUDYING THE RATES OF REACTION OF DISULFIDES WITH PROTEIN THIOL-GROUPS APPLIED TO BOVINE SERUM-ALBUMIN JOURNAL OF THE AMERICAN CHEMICAL SOCIETY 102 : 359 1980 22. ZHANG H A kinetic approach to characterize the electrostatic environments of thiol groups in proteins BIOORGANIC CHEMISTRY 26 : 356 1998

PY - 2003/12

Y1 - 2003/12

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AB - Sulphydryl-disulphide (SH-SS) exchange underlies many protein functions in processed foods. There is a need for reliable indicators of SH-SS exchange capacity in protein ingredients. An index for SH-SS exchange (SEI) is described using beta-lactoglobulin (BLG) or bovine serum albumin (BSA). The proteins were reacted with 2-pyridine disulphide (PDS Aldrithiol-2(TM)) or Ellman's reagent (DTNB) in a buffered medium (0.05 M phosphate buffer, pH 6.9) at 25degreesC. The 2nd order rate constant for protein SH-SS exchange with PDS or DTNB (k, M-1 s(-1)) was normalized by dividing by the rate constant for glutathione (GSH) reaction with PDS or DTNB (k*, M-1 s(-1)) determined under identical conditions. The capacity or SH-SS exchange was inversely related to SEI (= -log k/k*) values of 4.11 for BLG and 1.05 for BSA, based on measurements using PDS. Studies using DTNB yield SEI equal to 4.28 for BLG and 2.20 for BSA. The SH group of BSA was 100-1052 times more reactive than the SH group of BLG. In a medium containing 1.2 M sodium chloride the difference in SH-SS exchange capacity was similar to9000 fold. Differential scanning calorimetry (DSC) results showed that the conformational stability of BLG increased much more substantially than BSA when both proteins were exposed to 0.1-1.2 M sodium chloride concentrations. (C) 2003 Elsevier Ltd. All rights reserved.

KW - STABILITY FUNCTION RELATIONS

KW - CONFORMATIONAL STABILITY

KW - THIOL-GROUPS

KW - GELATION

KW - SALTS

U2 - 10.1016/S0308-8146(03)00150-X

DO - 10.1016/S0308-8146(03)00150-X

M3 - Article

VL - 83

SP - 541

EP - 545

IS - 4

ER -