Polymerization of Horseradish Peroxidase by a Laccase-Catalyzed Tyrosine Coupling Reaction

Dani Permana, Kosuke Minamihata, Tsuneyuki Tatsuke, Jae M. Lee, Takahiro Kusakabe, Masahiro Goto, Noriho Kamiya

研究成果: ジャーナルへの寄稿記事

1 引用 (Scopus)

抄録

The polymerization of proteins can create newly active and large bio-macromolecular assemblies that exhibit unique functionalities depending on the properties of the building block proteins and the protein units in polymers. Herein, the first enzymatic polymerization of horseradish peroxidase (HRP) is reported. Recombinant HRPs fused with a tyrosine-tag (Y-tag) through a flexible linker at the N- and/or C-termini are expressed in silkworm, Bombyx mori. Trametes sp. laccase (TL) is used to activate the tyrosine of Y-tagged HRPs with molecular O2 to form a tyrosyl-free radical, which initiates the tyrosine coupling reaction between the HRP units. A covalent dityrosine linkage is also formed through a HRP-catalyzed self-crosslinking reaction in the presence of H2O2. The addition of H2O2 in the self-polymerization of Y-tagged HRPs results in lower activity of the HRP polymers, whereas TL provides site-selectivity, mild reaction conditions and maintains the activity of the polymeric products. The cocrosslinking of Y-tagged HRPs and HRP-protein G (Y-HRP-pG) units catalyzed by TL shows a higher signal in enzyme-linked immunosorbent assay (ELISA) than the genetically pG-fused HRP, Y-HRP-pG, and its polymers. This new enzymatic polymerization of HRP promises to provide highly active and functionalized polymers for biomedical applications and diagnostics probes.

元の言語英語
記事番号1800531
ジャーナルBiotechnology Journal
14
発行部数6
DOI
出版物ステータス出版済み - 6 1 2019

Fingerprint

Laccase
Horseradish Peroxidase
Polymerization
Tyrosine
Polymers
Bombyx
Proteins
Trametes
Free Radicals
Enzyme-Linked Immunosorbent Assay

All Science Journal Classification (ASJC) codes

  • Applied Microbiology and Biotechnology
  • Molecular Medicine

これを引用

Polymerization of Horseradish Peroxidase by a Laccase-Catalyzed Tyrosine Coupling Reaction. / Permana, Dani; Minamihata, Kosuke; Tatsuke, Tsuneyuki; Lee, Jae M.; Kusakabe, Takahiro; Goto, Masahiro; Kamiya, Noriho.

:: Biotechnology Journal, 巻 14, 番号 6, 1800531, 01.06.2019.

研究成果: ジャーナルへの寄稿記事

@article{2d42fc3458ab456f8c107809dd149363,
title = "Polymerization of Horseradish Peroxidase by a Laccase-Catalyzed Tyrosine Coupling Reaction",
abstract = "The polymerization of proteins can create newly active and large bio-macromolecular assemblies that exhibit unique functionalities depending on the properties of the building block proteins and the protein units in polymers. Herein, the first enzymatic polymerization of horseradish peroxidase (HRP) is reported. Recombinant HRPs fused with a tyrosine-tag (Y-tag) through a flexible linker at the N- and/or C-termini are expressed in silkworm, Bombyx mori. Trametes sp. laccase (TL) is used to activate the tyrosine of Y-tagged HRPs with molecular O2 to form a tyrosyl-free radical, which initiates the tyrosine coupling reaction between the HRP units. A covalent dityrosine linkage is also formed through a HRP-catalyzed self-crosslinking reaction in the presence of H2O2. The addition of H2O2 in the self-polymerization of Y-tagged HRPs results in lower activity of the HRP polymers, whereas TL provides site-selectivity, mild reaction conditions and maintains the activity of the polymeric products. The cocrosslinking of Y-tagged HRPs and HRP-protein G (Y-HRP-pG) units catalyzed by TL shows a higher signal in enzyme-linked immunosorbent assay (ELISA) than the genetically pG-fused HRP, Y-HRP-pG, and its polymers. This new enzymatic polymerization of HRP promises to provide highly active and functionalized polymers for biomedical applications and diagnostics probes.",
author = "Dani Permana and Kosuke Minamihata and Tsuneyuki Tatsuke and Lee, {Jae M.} and Takahiro Kusakabe and Masahiro Goto and Noriho Kamiya",
year = "2019",
month = "6",
day = "1",
doi = "10.1002/biot.201800531",
language = "English",
volume = "14",
journal = "Biotechnology Journal",
issn = "1860-6768",
publisher = "Wiley-VCH Verlag",
number = "6",

}

TY - JOUR

T1 - Polymerization of Horseradish Peroxidase by a Laccase-Catalyzed Tyrosine Coupling Reaction

AU - Permana, Dani

AU - Minamihata, Kosuke

AU - Tatsuke, Tsuneyuki

AU - Lee, Jae M.

AU - Kusakabe, Takahiro

AU - Goto, Masahiro

AU - Kamiya, Noriho

PY - 2019/6/1

Y1 - 2019/6/1

N2 - The polymerization of proteins can create newly active and large bio-macromolecular assemblies that exhibit unique functionalities depending on the properties of the building block proteins and the protein units in polymers. Herein, the first enzymatic polymerization of horseradish peroxidase (HRP) is reported. Recombinant HRPs fused with a tyrosine-tag (Y-tag) through a flexible linker at the N- and/or C-termini are expressed in silkworm, Bombyx mori. Trametes sp. laccase (TL) is used to activate the tyrosine of Y-tagged HRPs with molecular O2 to form a tyrosyl-free radical, which initiates the tyrosine coupling reaction between the HRP units. A covalent dityrosine linkage is also formed through a HRP-catalyzed self-crosslinking reaction in the presence of H2O2. The addition of H2O2 in the self-polymerization of Y-tagged HRPs results in lower activity of the HRP polymers, whereas TL provides site-selectivity, mild reaction conditions and maintains the activity of the polymeric products. The cocrosslinking of Y-tagged HRPs and HRP-protein G (Y-HRP-pG) units catalyzed by TL shows a higher signal in enzyme-linked immunosorbent assay (ELISA) than the genetically pG-fused HRP, Y-HRP-pG, and its polymers. This new enzymatic polymerization of HRP promises to provide highly active and functionalized polymers for biomedical applications and diagnostics probes.

AB - The polymerization of proteins can create newly active and large bio-macromolecular assemblies that exhibit unique functionalities depending on the properties of the building block proteins and the protein units in polymers. Herein, the first enzymatic polymerization of horseradish peroxidase (HRP) is reported. Recombinant HRPs fused with a tyrosine-tag (Y-tag) through a flexible linker at the N- and/or C-termini are expressed in silkworm, Bombyx mori. Trametes sp. laccase (TL) is used to activate the tyrosine of Y-tagged HRPs with molecular O2 to form a tyrosyl-free radical, which initiates the tyrosine coupling reaction between the HRP units. A covalent dityrosine linkage is also formed through a HRP-catalyzed self-crosslinking reaction in the presence of H2O2. The addition of H2O2 in the self-polymerization of Y-tagged HRPs results in lower activity of the HRP polymers, whereas TL provides site-selectivity, mild reaction conditions and maintains the activity of the polymeric products. The cocrosslinking of Y-tagged HRPs and HRP-protein G (Y-HRP-pG) units catalyzed by TL shows a higher signal in enzyme-linked immunosorbent assay (ELISA) than the genetically pG-fused HRP, Y-HRP-pG, and its polymers. This new enzymatic polymerization of HRP promises to provide highly active and functionalized polymers for biomedical applications and diagnostics probes.

UR - http://www.scopus.com/inward/record.url?scp=85064490040&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85064490040&partnerID=8YFLogxK

U2 - 10.1002/biot.201800531

DO - 10.1002/biot.201800531

M3 - Article

C2 - 30810287

AN - SCOPUS:85064490040

VL - 14

JO - Biotechnology Journal

JF - Biotechnology Journal

SN - 1860-6768

IS - 6

M1 - 1800531

ER -