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Site-selective immobilization of streptavidin on enzymatically biotinylated bacterial magnetic particles

Published online by Cambridge University Press:  01 February 2011

Yoshiaki Maeda ,
Tomoko Yoshino ,
Haruko Takeyama ,
Masaaki Takahashi ,
Harumi Ginya ,
Junko Asahina ,
Hideji Tajima  and
Tadashi Matsunaga
Yoshiaki Maeda
Affiliation:
[email protected], Tokyo University of Agriculture and Technology, Department of Biotechnology, 2-24-16, Naka-cho, Koganei, 184-8588, Japan
Tomoko Yoshino
Affiliation:
[email protected], Tokyo University of Agriculture and Technology, Department of Biotechnology, 2-24-16, Naka-cho, Koganei, 184-8588, Japan
Haruko Takeyama
Affiliation:
[email protected], Tokyo University of Agriculture and Technology, Department of Biotechnology, 2-24-16, Naka-cho, Koganei, 184-8588, Japan
Masaaki Takahashi
Affiliation:
[email protected], Precision System Science Co., Ltd., Japan, 88 Kamihongo, Matsudo, Chiba, 271-0064, Japan
Harumi Ginya
Affiliation:
[email protected], Precision System Science Co., Ltd., Japan, 88 Kamihongo, Matsudo, Chiba, 271-0064, Japan
Junko Asahina
Affiliation:
[email protected], Precision System Science Co., Ltd., Japan, 88 Kamihongo, Matsudo, Chiba, 271-0064, Japan
Hideji Tajima
Affiliation:
[email protected], Precision System Science Co., Ltd., Japan, 88 Kamihongo, Matsudo, Chiba, 271-0064, Japan
Tadashi Matsunaga
Affiliation:
[email protected], Tokyo University of Agriculture and Technology, Department of Biotechnology, 2-24-16, Naka-cho, Koganei, 184-8588, Japan
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Abstract

Biotinylated magnetic nanoparticles were constructed by displaying biotin acceptor peptide (BAP) on the surfaces of bacterial magnetic particles (BacMPs) synthesized by Magnetospirillum magneticum AMB-1. Both BAP and green fluorescent protein (GFP) were fused to Mms13 that was isolated from BacMP membranes. The localization of the fusion protein, BAP-Mms13-GFP, was confirmed by fluorescence analysis. BacMPs that expressed BAP-Mms13-GFP (BAP/GFP-BacMPs) were extracted from bacterial cells and incubated with biotin and Escherichia coli biotin ligase. The in vitro biotinylation of BAP/GFP-BacMPs was confirmed using alkaline phosphatase (ALP)-streptavidin detection. The conjugation system developed in this study provides a method for producing biotin- or streptavidin-labeled magnetic nanoparticles without the use of a crosslinker reagent. Various functional materials can be immobilized site-selectively onto these uniquely designed BacMPs. By combining this site-selective biotinylation technology and protein display methodology, increasingly innovative and attractive magnetic nano-materials can be constructed.

Keywords

biomaterialmagneticparticulate
Type
Research Article
Information
MRS Online Proceedings Library (OPL) , Volume 1094: Symposium DD – From Biological Materials to Biomimetic Material Synthesis , 2008 , 1094-DD07-19
Copyright
Copyright © Materials Research Society 2008

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