Formation of protein-oligosaccharide conjugates by laccase and tyrosinase

Emilia Selinheimo (Corresponding Author), Piritta Lampila, Maija-Liisa Mattinen, Johanna Buchert

Research output: Contribution to journalArticleScientificpeer-review

90 Citations (Scopus)

Abstract

Proteins and certain carbohydrates contain phenolic moieties, which are potential sites for modification of the function of the biopolymers. In this study, the capability of two different fungal oxidative enzymes, laccase from Trametes hirsuta (ThL) and tyrosinase from Trichoderma reesei (TrT), to catalyze formation of hetero-cross-linking between tyrosine side chains of α-casein and phenolic acids of hydrolyzed oat spelt xylan (hOSX) was studied. Formation of reaction products was followed by size exclusion chromatography (SEC), fluorescence spectroscopy, and SDS-PAGE, using specific staining methods for proteins and protein−carbohydrate conjugates. ThL and TrT were observed to differ significantly in their ability to catalyze the formation of protein−carbohydrate conjugates or the linking of the small molecular weight phenolic compounds to α-casein. The efficiency of these enzymes to directly cross-link protein also differed notably. TrT was able to cross-link α-casein more efficiently than ThL. ThL-catalyzed casein cross-linking was significantly enhanced by ferulic acid, p-coumaric acid, and also hOSX. The main reaction products by ThL appeared to be phenolic acid-bridged α-caseins. Indications of hetero-cross-link formation between α-casein and hOSX by both oxidative enzymes could be visualized by glycoprotein-specific staining in the SDS-PAGE analysis, although ThL was observed to be more effective in the heteroconjugate formation than TrT.
Original languageEnglish
Pages (from-to)3118 - 3128
Number of pages11
JournalJournal of Agricultural and Food Chemistry
Volume56
Issue number9
DOIs
Publication statusPublished - 2008
MoE publication typeA1 Journal article-refereed

Fingerprint

Laccase
Monophenol Monooxygenase
laccase
Caseins
Oligosaccharides
catechol oxidase
oligosaccharides
casein
Triticum aestivum subsp. spelta
Xylans
xylan
oats
Proteins
ferulic acid
proteins
phenolic acids
Reaction products
crosslinking
Polyacrylamide Gel Electrophoresis
fluorescence emission spectroscopy

Keywords

  • Laccase
  • tyrosinase
  • protein
  • xylan
  • phenolic acids
  • cross-linking

Cite this

Selinheimo, Emilia ; Lampila, Piritta ; Mattinen, Maija-Liisa ; Buchert, Johanna. / Formation of protein-oligosaccharide conjugates by laccase and tyrosinase. In: Journal of Agricultural and Food Chemistry. 2008 ; Vol. 56, No. 9. pp. 3118 - 3128.
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abstract = "Proteins and certain carbohydrates contain phenolic moieties, which are potential sites for modification of the function of the biopolymers. In this study, the capability of two different fungal oxidative enzymes, laccase from Trametes hirsuta (ThL) and tyrosinase from Trichoderma reesei (TrT), to catalyze formation of hetero-cross-linking between tyrosine side chains of α-casein and phenolic acids of hydrolyzed oat spelt xylan (hOSX) was studied. Formation of reaction products was followed by size exclusion chromatography (SEC), fluorescence spectroscopy, and SDS-PAGE, using specific staining methods for proteins and protein−carbohydrate conjugates. ThL and TrT were observed to differ significantly in their ability to catalyze the formation of protein−carbohydrate conjugates or the linking of the small molecular weight phenolic compounds to α-casein. The efficiency of these enzymes to directly cross-link protein also differed notably. TrT was able to cross-link α-casein more efficiently than ThL. ThL-catalyzed casein cross-linking was significantly enhanced by ferulic acid, p-coumaric acid, and also hOSX. The main reaction products by ThL appeared to be phenolic acid-bridged α-caseins. Indications of hetero-cross-link formation between α-casein and hOSX by both oxidative enzymes could be visualized by glycoprotein-specific staining in the SDS-PAGE analysis, although ThL was observed to be more effective in the heteroconjugate formation than TrT.",
keywords = "Laccase, tyrosinase, protein, xylan, phenolic acids, cross-linking",
author = "Emilia Selinheimo and Piritta Lampila and Maija-Liisa Mattinen and Johanna Buchert",
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volume = "56",
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journal = "Journal of Agricultural and Food Chemistry",
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publisher = "American Chemical Society",
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Selinheimo, E, Lampila, P, Mattinen, M-L & Buchert, J 2008, 'Formation of protein-oligosaccharide conjugates by laccase and tyrosinase', Journal of Agricultural and Food Chemistry, vol. 56, no. 9, pp. 3118 - 3128. https://doi.org/10.1021/jf0730791

Formation of protein-oligosaccharide conjugates by laccase and tyrosinase. / Selinheimo, Emilia (Corresponding Author); Lampila, Piritta; Mattinen, Maija-Liisa; Buchert, Johanna.

In: Journal of Agricultural and Food Chemistry, Vol. 56, No. 9, 2008, p. 3118 - 3128.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Formation of protein-oligosaccharide conjugates by laccase and tyrosinase

AU - Selinheimo, Emilia

AU - Lampila, Piritta

AU - Mattinen, Maija-Liisa

AU - Buchert, Johanna

PY - 2008

Y1 - 2008

N2 - Proteins and certain carbohydrates contain phenolic moieties, which are potential sites for modification of the function of the biopolymers. In this study, the capability of two different fungal oxidative enzymes, laccase from Trametes hirsuta (ThL) and tyrosinase from Trichoderma reesei (TrT), to catalyze formation of hetero-cross-linking between tyrosine side chains of α-casein and phenolic acids of hydrolyzed oat spelt xylan (hOSX) was studied. Formation of reaction products was followed by size exclusion chromatography (SEC), fluorescence spectroscopy, and SDS-PAGE, using specific staining methods for proteins and protein−carbohydrate conjugates. ThL and TrT were observed to differ significantly in their ability to catalyze the formation of protein−carbohydrate conjugates or the linking of the small molecular weight phenolic compounds to α-casein. The efficiency of these enzymes to directly cross-link protein also differed notably. TrT was able to cross-link α-casein more efficiently than ThL. ThL-catalyzed casein cross-linking was significantly enhanced by ferulic acid, p-coumaric acid, and also hOSX. The main reaction products by ThL appeared to be phenolic acid-bridged α-caseins. Indications of hetero-cross-link formation between α-casein and hOSX by both oxidative enzymes could be visualized by glycoprotein-specific staining in the SDS-PAGE analysis, although ThL was observed to be more effective in the heteroconjugate formation than TrT.

AB - Proteins and certain carbohydrates contain phenolic moieties, which are potential sites for modification of the function of the biopolymers. In this study, the capability of two different fungal oxidative enzymes, laccase from Trametes hirsuta (ThL) and tyrosinase from Trichoderma reesei (TrT), to catalyze formation of hetero-cross-linking between tyrosine side chains of α-casein and phenolic acids of hydrolyzed oat spelt xylan (hOSX) was studied. Formation of reaction products was followed by size exclusion chromatography (SEC), fluorescence spectroscopy, and SDS-PAGE, using specific staining methods for proteins and protein−carbohydrate conjugates. ThL and TrT were observed to differ significantly in their ability to catalyze the formation of protein−carbohydrate conjugates or the linking of the small molecular weight phenolic compounds to α-casein. The efficiency of these enzymes to directly cross-link protein also differed notably. TrT was able to cross-link α-casein more efficiently than ThL. ThL-catalyzed casein cross-linking was significantly enhanced by ferulic acid, p-coumaric acid, and also hOSX. The main reaction products by ThL appeared to be phenolic acid-bridged α-caseins. Indications of hetero-cross-link formation between α-casein and hOSX by both oxidative enzymes could be visualized by glycoprotein-specific staining in the SDS-PAGE analysis, although ThL was observed to be more effective in the heteroconjugate formation than TrT.

KW - Laccase

KW - tyrosinase

KW - protein

KW - xylan

KW - phenolic acids

KW - cross-linking

U2 - 10.1021/jf0730791

DO - 10.1021/jf0730791

M3 - Article

VL - 56

SP - 3118

EP - 3128

JO - Journal of Agricultural and Food Chemistry

JF - Journal of Agricultural and Food Chemistry

SN - 0021-8561

IS - 9

ER -