Enzymatic cross-linking of ß-lactoglobulin in solution and at air-water interface: Structural constraints

Dilek Ercili-Cura (Corresponding Author), Riitta Partanen, F. Husband, M. Ridout, A. Macierzanka, Martina Lille, Harry Boer, Raija Lantto, Johanna Buchert, A. Mackie

Research output: Contribution to journalArticleScientificpeer-review

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Abstract

Effective and controlled use of cross-linking enzymes in structure engineering of food systems depends on characterization of the favorable conditions for enzyme-substrate complex and the limiting factors for the desired modification. In this respect, we analyzed the susceptibility of bovine β-lactoglobulin (BLG) to enzymatic cross-linking by Trichoderma reesei tyrosinase (TrTyr) and transglutaminase (TG). Changes in BLG molecular structure were determined at pH 6.8, 7.5 and 9.0 before and after high-temperature heat treatment. The conformational change was linked to efficiency of protein cross-linking. BLG was not susceptible to TrTyr without heat treatment. TG, however, induced inter-molecular cross-links at pH 7.5 and 9.0. After the heat treatments, BLG molecules adopted a molten-globule-like conformation. Both of the enzymes were able to form inter-molecular cross-links between heat-denatured BLG molecules. Electrophoretic mobility and broadness of the oligomer bands created by both enzymes on SDS-PAGE gels showed differences which were linked to the availability and number of target amino acid residues. Evidence for intra-molecular cross-linking was obtained. Once adsorbed to air/water interface, BLG formed a viscoelastic surface film which was characterized by surface shear rheology. Application of cross-linking enzymes under a dense layer of BLG molecules at the interface led to decreasing G′ with time. Intra-molecular links were most probably favored against inter-molecular on packed BLG layer leading to constrained molecules. Results in general emphasize the importance of structural and colloidal aspects of protein molecules in controlling inter/intra-molecular bond formation by cross-linking enzymes.

Original languageEnglish
Pages (from-to)1-9
Number of pages9
JournalFood Hydrocolloids
Volume28
Issue number1
DOIs
Publication statusPublished - 2012
MoE publication typeA1 Journal article-refereed

Fingerprint

lactoglobulins
Lactoglobulins
crosslinking
Enzymes
Air
air
Water
cattle
Molecules
Hot Temperature
water
Heat treatment
enzymes
Trichoderma reesei
Trichoderma
Transglutaminases
Monophenol Monooxygenase
protein-glutamine gamma-glutamyltransferase
heat treatment
Proteins

Keywords

  • ß-Lactoglobulin
  • tyrosinase
  • transglutaminase
  • cross-linking
  • interface

Cite this

Ercili-Cura, Dilek ; Partanen, Riitta ; Husband, F. ; Ridout, M. ; Macierzanka, A. ; Lille, Martina ; Boer, Harry ; Lantto, Raija ; Buchert, Johanna ; Mackie, A. / Enzymatic cross-linking of ß-lactoglobulin in solution and at air-water interface : Structural constraints. In: Food Hydrocolloids. 2012 ; Vol. 28, No. 1. pp. 1-9.
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abstract = "Effective and controlled use of cross-linking enzymes in structure engineering of food systems depends on characterization of the favorable conditions for enzyme-substrate complex and the limiting factors for the desired modification. In this respect, we analyzed the susceptibility of bovine β-lactoglobulin (BLG) to enzymatic cross-linking by Trichoderma reesei tyrosinase (TrTyr) and transglutaminase (TG). Changes in BLG molecular structure were determined at pH 6.8, 7.5 and 9.0 before and after high-temperature heat treatment. The conformational change was linked to efficiency of protein cross-linking. BLG was not susceptible to TrTyr without heat treatment. TG, however, induced inter-molecular cross-links at pH 7.5 and 9.0. After the heat treatments, BLG molecules adopted a molten-globule-like conformation. Both of the enzymes were able to form inter-molecular cross-links between heat-denatured BLG molecules. Electrophoretic mobility and broadness of the oligomer bands created by both enzymes on SDS-PAGE gels showed differences which were linked to the availability and number of target amino acid residues. Evidence for intra-molecular cross-linking was obtained. Once adsorbed to air/water interface, BLG formed a viscoelastic surface film which was characterized by surface shear rheology. Application of cross-linking enzymes under a dense layer of BLG molecules at the interface led to decreasing G′ with time. Intra-molecular links were most probably favored against inter-molecular on packed BLG layer leading to constrained molecules. Results in general emphasize the importance of structural and colloidal aspects of protein molecules in controlling inter/intra-molecular bond formation by cross-linking enzymes.",
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Enzymatic cross-linking of ß-lactoglobulin in solution and at air-water interface : Structural constraints. / Ercili-Cura, Dilek (Corresponding Author); Partanen, Riitta; Husband, F.; Ridout, M.; Macierzanka, A.; Lille, Martina; Boer, Harry; Lantto, Raija; Buchert, Johanna; Mackie, A.

In: Food Hydrocolloids, Vol. 28, No. 1, 2012, p. 1-9.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Enzymatic cross-linking of ß-lactoglobulin in solution and at air-water interface

T2 - Structural constraints

AU - Ercili-Cura, Dilek

AU - Partanen, Riitta

AU - Husband, F.

AU - Ridout, M.

AU - Macierzanka, A.

AU - Lille, Martina

AU - Boer, Harry

AU - Lantto, Raija

AU - Buchert, Johanna

AU - Mackie, A.

PY - 2012

Y1 - 2012

N2 - Effective and controlled use of cross-linking enzymes in structure engineering of food systems depends on characterization of the favorable conditions for enzyme-substrate complex and the limiting factors for the desired modification. In this respect, we analyzed the susceptibility of bovine β-lactoglobulin (BLG) to enzymatic cross-linking by Trichoderma reesei tyrosinase (TrTyr) and transglutaminase (TG). Changes in BLG molecular structure were determined at pH 6.8, 7.5 and 9.0 before and after high-temperature heat treatment. The conformational change was linked to efficiency of protein cross-linking. BLG was not susceptible to TrTyr without heat treatment. TG, however, induced inter-molecular cross-links at pH 7.5 and 9.0. After the heat treatments, BLG molecules adopted a molten-globule-like conformation. Both of the enzymes were able to form inter-molecular cross-links between heat-denatured BLG molecules. Electrophoretic mobility and broadness of the oligomer bands created by both enzymes on SDS-PAGE gels showed differences which were linked to the availability and number of target amino acid residues. Evidence for intra-molecular cross-linking was obtained. Once adsorbed to air/water interface, BLG formed a viscoelastic surface film which was characterized by surface shear rheology. Application of cross-linking enzymes under a dense layer of BLG molecules at the interface led to decreasing G′ with time. Intra-molecular links were most probably favored against inter-molecular on packed BLG layer leading to constrained molecules. Results in general emphasize the importance of structural and colloidal aspects of protein molecules in controlling inter/intra-molecular bond formation by cross-linking enzymes.

AB - Effective and controlled use of cross-linking enzymes in structure engineering of food systems depends on characterization of the favorable conditions for enzyme-substrate complex and the limiting factors for the desired modification. In this respect, we analyzed the susceptibility of bovine β-lactoglobulin (BLG) to enzymatic cross-linking by Trichoderma reesei tyrosinase (TrTyr) and transglutaminase (TG). Changes in BLG molecular structure were determined at pH 6.8, 7.5 and 9.0 before and after high-temperature heat treatment. The conformational change was linked to efficiency of protein cross-linking. BLG was not susceptible to TrTyr without heat treatment. TG, however, induced inter-molecular cross-links at pH 7.5 and 9.0. After the heat treatments, BLG molecules adopted a molten-globule-like conformation. Both of the enzymes were able to form inter-molecular cross-links between heat-denatured BLG molecules. Electrophoretic mobility and broadness of the oligomer bands created by both enzymes on SDS-PAGE gels showed differences which were linked to the availability and number of target amino acid residues. Evidence for intra-molecular cross-linking was obtained. Once adsorbed to air/water interface, BLG formed a viscoelastic surface film which was characterized by surface shear rheology. Application of cross-linking enzymes under a dense layer of BLG molecules at the interface led to decreasing G′ with time. Intra-molecular links were most probably favored against inter-molecular on packed BLG layer leading to constrained molecules. Results in general emphasize the importance of structural and colloidal aspects of protein molecules in controlling inter/intra-molecular bond formation by cross-linking enzymes.

KW - ß-Lactoglobulin

KW - tyrosinase

KW - transglutaminase

KW - cross-linking

KW - interface

U2 - 10.1016/j.foodhyd.2011.11.010

DO - 10.1016/j.foodhyd.2011.11.010

M3 - Article

VL - 28

SP - 1

EP - 9

JO - Food Hydrocolloids

JF - Food Hydrocolloids

SN - 0268-005X

IS - 1

ER -