Cross-linking of β-casein by Trichoderma reesei tyrosinase and Streptoverticillium mobaraense transglutaminase followed by SEC–MALLS

Evanthia Monogioudi (Corresponding Author), Nathalie Creusot, Kristiina Kruus, Harry Gruppen, Johanna Buchert, Maija-Liisa Mattinen

Research output: Contribution to journalArticleScientificpeer-review

31 Citations (Scopus)

Abstract

Enzymatic modification of proteins, in order to produce functional materials such as hydrogels, adhesives and films via cross-linked networks or scaffolds of proteins, is a constantly evolving technology to create tailored micro- and nanostructured materials for food, cosmetic, and medical applications. For the successful utilization of oxidoreductases or transferases such as tyrosinases and transglutaminases, respectively, it is crucial to understand the action of these enzymes on protein substrates. In this study, cross-linking of the milk protein β-casein by Trichoderma reesei tyrosinase (TrTyr) was studied using size-exclusion chromatography (SEC) equipped with multi-angle light scattering (MALLS) and ultraviolet/visible (UV/Vis) detectors in order to determine the molecular mass (MM), radius of gyration (RG) and degree of polymerization (DP) of the reaction products. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) was used to detect early polymerization states. The widely used Streptoverticillium mobaraense transglutaminase (Tgase) was used for comparison to tyrosinase from T. reesei. The results showed that cross-linking of β-casein by these two different types of enzymes resulted in the formation of polymerized reaction products with MM ranging from 500 to 1700 kg mol−1 depending on the enzyme dosage and incubation time. The DP varied from 21 to 71, respectively. In the case of TrTyr the polymerized reaction products were slightly colored, and formation of the covalent cross-linking of β-casein could be monitored by UV/Vis as a function of incubation time.
Original languageEnglish
Pages (from-to)2008 - 2015
Number of pages8
JournalFood Hydrocolloids
Volume23
Issue number7
DOIs
Publication statusPublished - 2009
MoE publication typeA1 Journal article-refereed

Fingerprint

Streptomyces mobaraensis
Trichoderma reesei
Casein
Trichoderma
Transglutaminases
Monophenol Monooxygenase
protein-glutamine gamma-glutamyltransferase
Caseins
Reaction products
crosslinking
polymerization
casein
Enzymes
Polymerization
Molecular mass
Proteins
enzymes
scaffolding proteins
molecular weight
nanomaterials

Keywords

  • Tyrosinase
  • Transglutaminase
  • Cross-linking
  • beta-Casein
  • Molecular mass
  • Degree of polymerization
  • Color formation

Cite this

Monogioudi, Evanthia ; Creusot, Nathalie ; Kruus, Kristiina ; Gruppen, Harry ; Buchert, Johanna ; Mattinen, Maija-Liisa. / Cross-linking of β-casein by Trichoderma reesei tyrosinase and Streptoverticillium mobaraense transglutaminase followed by SEC–MALLS. In: Food Hydrocolloids. 2009 ; Vol. 23, No. 7. pp. 2008 - 2015.
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title = "Cross-linking of β-casein by Trichoderma reesei tyrosinase and Streptoverticillium mobaraense transglutaminase followed by SEC–MALLS",
abstract = "Enzymatic modification of proteins, in order to produce functional materials such as hydrogels, adhesives and films via cross-linked networks or scaffolds of proteins, is a constantly evolving technology to create tailored micro- and nanostructured materials for food, cosmetic, and medical applications. For the successful utilization of oxidoreductases or transferases such as tyrosinases and transglutaminases, respectively, it is crucial to understand the action of these enzymes on protein substrates. In this study, cross-linking of the milk protein β-casein by Trichoderma reesei tyrosinase (TrTyr) was studied using size-exclusion chromatography (SEC) equipped with multi-angle light scattering (MALLS) and ultraviolet/visible (UV/Vis) detectors in order to determine the molecular mass (MM), radius of gyration (RG) and degree of polymerization (DP) of the reaction products. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) was used to detect early polymerization states. The widely used Streptoverticillium mobaraense transglutaminase (Tgase) was used for comparison to tyrosinase from T. reesei. The results showed that cross-linking of β-casein by these two different types of enzymes resulted in the formation of polymerized reaction products with MM ranging from 500 to 1700 kg mol−1 depending on the enzyme dosage and incubation time. The DP varied from 21 to 71, respectively. In the case of TrTyr the polymerized reaction products were slightly colored, and formation of the covalent cross-linking of β-casein could be monitored by UV/Vis as a function of incubation time.",
keywords = "Tyrosinase, Transglutaminase, Cross-linking, beta-Casein, Molecular mass, Degree of polymerization, Color formation",
author = "Evanthia Monogioudi and Nathalie Creusot and Kristiina Kruus and Harry Gruppen and Johanna Buchert and Maija-Liisa Mattinen",
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doi = "10.1016/j.foodhyd.2009.03.011",
language = "English",
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Cross-linking of β-casein by Trichoderma reesei tyrosinase and Streptoverticillium mobaraense transglutaminase followed by SEC–MALLS. / Monogioudi, Evanthia (Corresponding Author); Creusot, Nathalie; Kruus, Kristiina; Gruppen, Harry; Buchert, Johanna; Mattinen, Maija-Liisa.

In: Food Hydrocolloids, Vol. 23, No. 7, 2009, p. 2008 - 2015.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Cross-linking of β-casein by Trichoderma reesei tyrosinase and Streptoverticillium mobaraense transglutaminase followed by SEC–MALLS

AU - Monogioudi, Evanthia

AU - Creusot, Nathalie

AU - Kruus, Kristiina

AU - Gruppen, Harry

AU - Buchert, Johanna

AU - Mattinen, Maija-Liisa

PY - 2009

Y1 - 2009

N2 - Enzymatic modification of proteins, in order to produce functional materials such as hydrogels, adhesives and films via cross-linked networks or scaffolds of proteins, is a constantly evolving technology to create tailored micro- and nanostructured materials for food, cosmetic, and medical applications. For the successful utilization of oxidoreductases or transferases such as tyrosinases and transglutaminases, respectively, it is crucial to understand the action of these enzymes on protein substrates. In this study, cross-linking of the milk protein β-casein by Trichoderma reesei tyrosinase (TrTyr) was studied using size-exclusion chromatography (SEC) equipped with multi-angle light scattering (MALLS) and ultraviolet/visible (UV/Vis) detectors in order to determine the molecular mass (MM), radius of gyration (RG) and degree of polymerization (DP) of the reaction products. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) was used to detect early polymerization states. The widely used Streptoverticillium mobaraense transglutaminase (Tgase) was used for comparison to tyrosinase from T. reesei. The results showed that cross-linking of β-casein by these two different types of enzymes resulted in the formation of polymerized reaction products with MM ranging from 500 to 1700 kg mol−1 depending on the enzyme dosage and incubation time. The DP varied from 21 to 71, respectively. In the case of TrTyr the polymerized reaction products were slightly colored, and formation of the covalent cross-linking of β-casein could be monitored by UV/Vis as a function of incubation time.

AB - Enzymatic modification of proteins, in order to produce functional materials such as hydrogels, adhesives and films via cross-linked networks or scaffolds of proteins, is a constantly evolving technology to create tailored micro- and nanostructured materials for food, cosmetic, and medical applications. For the successful utilization of oxidoreductases or transferases such as tyrosinases and transglutaminases, respectively, it is crucial to understand the action of these enzymes on protein substrates. In this study, cross-linking of the milk protein β-casein by Trichoderma reesei tyrosinase (TrTyr) was studied using size-exclusion chromatography (SEC) equipped with multi-angle light scattering (MALLS) and ultraviolet/visible (UV/Vis) detectors in order to determine the molecular mass (MM), radius of gyration (RG) and degree of polymerization (DP) of the reaction products. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) was used to detect early polymerization states. The widely used Streptoverticillium mobaraense transglutaminase (Tgase) was used for comparison to tyrosinase from T. reesei. The results showed that cross-linking of β-casein by these two different types of enzymes resulted in the formation of polymerized reaction products with MM ranging from 500 to 1700 kg mol−1 depending on the enzyme dosage and incubation time. The DP varied from 21 to 71, respectively. In the case of TrTyr the polymerized reaction products were slightly colored, and formation of the covalent cross-linking of β-casein could be monitored by UV/Vis as a function of incubation time.

KW - Tyrosinase

KW - Transglutaminase

KW - Cross-linking

KW - beta-Casein

KW - Molecular mass

KW - Degree of polymerization

KW - Color formation

U2 - 10.1016/j.foodhyd.2009.03.011

DO - 10.1016/j.foodhyd.2009.03.011

M3 - Article

VL - 23

SP - 2008

EP - 2015

JO - Food Hydrocolloids

JF - Food Hydrocolloids

SN - 0268-005X

IS - 7

ER -