Transglutaminase-mediated protein immobilization to casein nanolayers created on a plastic surface

Noriho Kamiya, Satoshi Doi, Jo Tominaga, Hirofumi Ichinose, Masahiro Goto

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

An enzymatic method for covalent and site-specific immobilization of recombinant proteins on a plastic surface was explored. Using Escherichia coli alkaline phosphatase (AP) with a specific peptide tag (MKHKGS) genetically incorporated at the N-terminus as a model (NK-AP), microbial transglutaminase (MTG)-mediated protein immobilization was demonstrated. To generate a reactive surface for MTG, a 96-well polystyrene microtiter plate was physically coated with casein, a good MTG substrate. Successful immobilization of recombinant AP to the nanolayer of casein on the surface of the microtiter plate was verified by the detection of enzymatic activity. Since little activity was observed when wild-type AP was used, immobilization of NK-AP was likely directed by the specific peptide tag. When polymeric casein prepared by MTG was used as a matrix on the plate, the loading capacity of AP was increased about 2-fold compared to when casein was used as the matrix. Transglutaminase-mediated site-specific posttranslational modification of proteins offers one way of generating a variety of protein-based solid formulations for biotechnological applications.

Original languageEnglish
Pages (from-to)35-38
Number of pages4
JournalBiomacromolecules
Volume6
Issue number1
DOIs
Publication statusPublished - Jan 1 2005

Fingerprint

Casein
Transglutaminases
Phosphatases
Caseins
Immobilization
Plastics
Alkaline Phosphatase
Proteins
Peptides
Recombinant proteins
Polystyrenes
Post Translational Protein Processing
Recombinant Proteins
Escherichia coli
Substrates

All Science Journal Classification (ASJC) codes

  • Bioengineering
  • Biomaterials
  • Polymers and Plastics
  • Materials Chemistry

Cite this

Transglutaminase-mediated protein immobilization to casein nanolayers created on a plastic surface. / Kamiya, Noriho; Doi, Satoshi; Tominaga, Jo; Ichinose, Hirofumi; Goto, Masahiro.

In: Biomacromolecules, Vol. 6, No. 1, 01.01.2005, p. 35-38.

Research output: Contribution to journalArticle

@article{28506a1f2d8443db862a509a27bb2526,
title = "Transglutaminase-mediated protein immobilization to casein nanolayers created on a plastic surface",
abstract = "An enzymatic method for covalent and site-specific immobilization of recombinant proteins on a plastic surface was explored. Using Escherichia coli alkaline phosphatase (AP) with a specific peptide tag (MKHKGS) genetically incorporated at the N-terminus as a model (NK-AP), microbial transglutaminase (MTG)-mediated protein immobilization was demonstrated. To generate a reactive surface for MTG, a 96-well polystyrene microtiter plate was physically coated with casein, a good MTG substrate. Successful immobilization of recombinant AP to the nanolayer of casein on the surface of the microtiter plate was verified by the detection of enzymatic activity. Since little activity was observed when wild-type AP was used, immobilization of NK-AP was likely directed by the specific peptide tag. When polymeric casein prepared by MTG was used as a matrix on the plate, the loading capacity of AP was increased about 2-fold compared to when casein was used as the matrix. Transglutaminase-mediated site-specific posttranslational modification of proteins offers one way of generating a variety of protein-based solid formulations for biotechnological applications.",
author = "Noriho Kamiya and Satoshi Doi and Jo Tominaga and Hirofumi Ichinose and Masahiro Goto",
year = "2005",
month = "1",
day = "1",
doi = "10.1021/bm0494895",
language = "English",
volume = "6",
pages = "35--38",
journal = "Biomacromolecules",
issn = "1525-7797",
publisher = "American Chemical Society",
number = "1",

}

TY - JOUR

T1 - Transglutaminase-mediated protein immobilization to casein nanolayers created on a plastic surface

AU - Kamiya, Noriho

AU - Doi, Satoshi

AU - Tominaga, Jo

AU - Ichinose, Hirofumi

AU - Goto, Masahiro

PY - 2005/1/1

Y1 - 2005/1/1

N2 - An enzymatic method for covalent and site-specific immobilization of recombinant proteins on a plastic surface was explored. Using Escherichia coli alkaline phosphatase (AP) with a specific peptide tag (MKHKGS) genetically incorporated at the N-terminus as a model (NK-AP), microbial transglutaminase (MTG)-mediated protein immobilization was demonstrated. To generate a reactive surface for MTG, a 96-well polystyrene microtiter plate was physically coated with casein, a good MTG substrate. Successful immobilization of recombinant AP to the nanolayer of casein on the surface of the microtiter plate was verified by the detection of enzymatic activity. Since little activity was observed when wild-type AP was used, immobilization of NK-AP was likely directed by the specific peptide tag. When polymeric casein prepared by MTG was used as a matrix on the plate, the loading capacity of AP was increased about 2-fold compared to when casein was used as the matrix. Transglutaminase-mediated site-specific posttranslational modification of proteins offers one way of generating a variety of protein-based solid formulations for biotechnological applications.

AB - An enzymatic method for covalent and site-specific immobilization of recombinant proteins on a plastic surface was explored. Using Escherichia coli alkaline phosphatase (AP) with a specific peptide tag (MKHKGS) genetically incorporated at the N-terminus as a model (NK-AP), microbial transglutaminase (MTG)-mediated protein immobilization was demonstrated. To generate a reactive surface for MTG, a 96-well polystyrene microtiter plate was physically coated with casein, a good MTG substrate. Successful immobilization of recombinant AP to the nanolayer of casein on the surface of the microtiter plate was verified by the detection of enzymatic activity. Since little activity was observed when wild-type AP was used, immobilization of NK-AP was likely directed by the specific peptide tag. When polymeric casein prepared by MTG was used as a matrix on the plate, the loading capacity of AP was increased about 2-fold compared to when casein was used as the matrix. Transglutaminase-mediated site-specific posttranslational modification of proteins offers one way of generating a variety of protein-based solid formulations for biotechnological applications.

UR - http://www.scopus.com/inward/record.url?scp=14044250023&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=14044250023&partnerID=8YFLogxK

U2 - 10.1021/bm0494895

DO - 10.1021/bm0494895

M3 - Article

C2 - 15638501

AN - SCOPUS:14044250023

VL - 6

SP - 35

EP - 38

JO - Biomacromolecules

JF - Biomacromolecules

SN - 1525-7797

IS - 1

ER -