Identification of the adhesive domain of AtaA from Acinetobacter sp. Tol 5 and its application in immobilizing Escherichia coli

Cell immobilization is an important technique for efficiently utilizing whole-cell biocatalysts. We previously invented a method for bacterial cell immobilization using AtaA, a trimeric autotransporter adhesin from the highly sticky bacterium sp. Tol 5. However, except for species, only one bacteriu...

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Published inFrontiers in bioengineering and biotechnology Vol. 10; p. 1095057
Main Authors Yoshimoto, Shogo, Aoki, Sota, Ohara, Yuki, Ishikawa, Masahito, Suzuki, Atsuo, Linke, Dirk, Lupas, Andrei N, Hori, Katsutoshi
Format Journal Article
LanguageEnglish
Norwegian
Published Switzerland Frontiers Media S.A 09.01.2023
Subjects
adhesion
autotransporter
bacteria
Bioengineering and Biotechnology
immobilization
protein engineering
autotransporter
protein engineering
immobilization
adhesion
bacteria
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Summary:Cell immobilization is an important technique for efficiently utilizing whole-cell biocatalysts. We previously invented a method for bacterial cell immobilization using AtaA, a trimeric autotransporter adhesin from the highly sticky bacterium sp. Tol 5. However, except for species, only one bacterium has been successfully immobilized using AtaA. This is probably because the heterologous expression of large AtaA (1 MDa), that is a homotrimer of polypeptide chains composed of 3,630 amino acids, is difficult. In this study, we identified the adhesive domain of AtaA and constructed a miniaturized AtaA (mini-AtaA) to improve the heterologous expression of In-frame deletion mutants were used to perform functional mapping, revealing that the N-terminal head domain is essential for the adhesive feature of AtaA. The mini-AtaA, which contains a homotrimer of polypeptide chains from 775 amino acids and lacks the unnecessary part for its adhesion, was properly expressed in and a larger amount of molecules was displayed on the cell surface than that of full-length AtaA (FL-AtaA). The immobilization ratio of cells expressing mini-AtaA on a polyurethane foam support was significantly higher compared to the cells with or without FL-AtaA expression, respectively. The expression of mini-AtaA in had little effect on the cell growth and the activity of another enzyme reflecting the production level, and the immobilized cells could be used for repetitive enzymatic reactions as a whole-cell catalyst.
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NFR/286467
Present address: Masahito Ishikawa, Department of Bioscience, Nagahama Institute of Bio-Science and Technology, Nagahama, Japan
These authors have contributed equally to this work
Edited by: Ye-Wang Zhang, Jiangsu University, China
Sota Aoki, Meidensha Corporation, Tokyo, Japan
Yuki Ohara, Friend Microbe Inc., Nagoya, Japan
Reviewed by: Jiandong Cui, Tianjin University of Science and Technology, China
This article was submitted to Bioprocess Engineering, a section of the journal Frontiers in Bioengineering and Biotechnology
Guang Zhao, Shandong University, China
ISSN:2296-4185
2296-4185
DOI:10.3389/fbioe.2022.1095057