Enhanced heterologous protein display on bacterial magnetic particles using a lon protease gene deletion mutant in Magnetospirillum magneticum AMB-1

Yuka Kanetsuki, Tsuyoshi Tanaka, Tadashi Matsunaga, Tomoko Yoshino

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Bacterial magnetic particles (BacMPs) produced by the magnetotactic bacterium Magnetospirillum magneticum AMB-1, are used as magnetic supports or carriers for a variety of biomedical and environmental applications. Although protein expression systems on BacMPs have been established in previous studies, the expression efficiency was dependent on the introduced protein sequences. Recombinant human proteins are often poorly expressed on BacMPs because of proteolytic degradation by endogenous proteases. We constructed a lon protease gene deletion mutant strain (Δlon) of M. magneticum AMB-1 by homologous recombination to increase the efficiency of functional protein display on BacMPs using Δlon host cells. Wild-type and Δlon-M. magneticum AMB-1 cells were transformed using expression plasmids for human proteins, thyroid-stimulating hormone receptor (TSHR) and the class II major histocompatibility complex (MHC II) molecules onto BacMPs. Although mRNA expression of both TSHR and MHC II was the same level in the wild-type and Δlon transformants, the protein expression levels in Δlon transformants were significantly increased versus wild-type cells. Furthermore, the amounts of two different human proteins on BacMPs were successfully improved. This phenomenon could be due to the reduction of the degradation of target proteins in the Δlon strain. This is the first report to construct a protease deletion mutant in magnetotactic bacteria. The Δlon strain is a useful host to provide BacMPs displaying target proteins for various experimental, and ultimately, clinical applications.

Original languageEnglish
Pages (from-to)65-70
Number of pages6
JournalJournal of Bioscience and Bioengineering
Volume116
Issue number1
DOIs
Publication statusPublished - 2013 Jul
Externally publishedYes

Fingerprint

Magnetospirillum
Protease La
Gene Deletion
Genes
Display devices
Proteins
Thyrotropin Receptors
Major Histocompatibility Complex
Peptide Hydrolases
Bacteria
Bacterial Proteins
Homologous Recombination
Recombinant Proteins
Proteolysis
Degradation
Plasmids
Messenger RNA

Keywords

  • Bacterial magnetic particles
  • Deletion mutant
  • Lon protease
  • Magnetotactic bacteria
  • Protein display

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology
  • Bioengineering

Cite this

Enhanced heterologous protein display on bacterial magnetic particles using a lon protease gene deletion mutant in Magnetospirillum magneticum AMB-1. / Kanetsuki, Yuka; Tanaka, Tsuyoshi; Matsunaga, Tadashi; Yoshino, Tomoko.

In: Journal of Bioscience and Bioengineering, Vol. 116, No. 1, 07.2013, p. 65-70.

Research output: Contribution to journalArticle

@article{7e6561ddb9354786b37aef38b381e19e,
title = "Enhanced heterologous protein display on bacterial magnetic particles using a lon protease gene deletion mutant in Magnetospirillum magneticum AMB-1",
abstract = "Bacterial magnetic particles (BacMPs) produced by the magnetotactic bacterium Magnetospirillum magneticum AMB-1, are used as magnetic supports or carriers for a variety of biomedical and environmental applications. Although protein expression systems on BacMPs have been established in previous studies, the expression efficiency was dependent on the introduced protein sequences. Recombinant human proteins are often poorly expressed on BacMPs because of proteolytic degradation by endogenous proteases. We constructed a lon protease gene deletion mutant strain (Δlon) of M. magneticum AMB-1 by homologous recombination to increase the efficiency of functional protein display on BacMPs using Δlon host cells. Wild-type and Δlon-M. magneticum AMB-1 cells were transformed using expression plasmids for human proteins, thyroid-stimulating hormone receptor (TSHR) and the class II major histocompatibility complex (MHC II) molecules onto BacMPs. Although mRNA expression of both TSHR and MHC II was the same level in the wild-type and Δlon transformants, the protein expression levels in Δlon transformants were significantly increased versus wild-type cells. Furthermore, the amounts of two different human proteins on BacMPs were successfully improved. This phenomenon could be due to the reduction of the degradation of target proteins in the Δlon strain. This is the first report to construct a protease deletion mutant in magnetotactic bacteria. The Δlon strain is a useful host to provide BacMPs displaying target proteins for various experimental, and ultimately, clinical applications.",
keywords = "Bacterial magnetic particles, Deletion mutant, Lon protease, Magnetotactic bacteria, Protein display",
author = "Yuka Kanetsuki and Tsuyoshi Tanaka and Tadashi Matsunaga and Tomoko Yoshino",
year = "2013",
month = "7",
doi = "10.1016/j.jbiosc.2013.01.016",
language = "English",
volume = "116",
pages = "65--70",
journal = "Journal of Bioscience and Bioengineering",
issn = "1389-1723",
publisher = "Elsevier",
number = "1",

}

TY - JOUR

T1 - Enhanced heterologous protein display on bacterial magnetic particles using a lon protease gene deletion mutant in Magnetospirillum magneticum AMB-1

AU - Kanetsuki, Yuka

AU - Tanaka, Tsuyoshi

AU - Matsunaga, Tadashi

AU - Yoshino, Tomoko

PY - 2013/7

Y1 - 2013/7

N2 - Bacterial magnetic particles (BacMPs) produced by the magnetotactic bacterium Magnetospirillum magneticum AMB-1, are used as magnetic supports or carriers for a variety of biomedical and environmental applications. Although protein expression systems on BacMPs have been established in previous studies, the expression efficiency was dependent on the introduced protein sequences. Recombinant human proteins are often poorly expressed on BacMPs because of proteolytic degradation by endogenous proteases. We constructed a lon protease gene deletion mutant strain (Δlon) of M. magneticum AMB-1 by homologous recombination to increase the efficiency of functional protein display on BacMPs using Δlon host cells. Wild-type and Δlon-M. magneticum AMB-1 cells were transformed using expression plasmids for human proteins, thyroid-stimulating hormone receptor (TSHR) and the class II major histocompatibility complex (MHC II) molecules onto BacMPs. Although mRNA expression of both TSHR and MHC II was the same level in the wild-type and Δlon transformants, the protein expression levels in Δlon transformants were significantly increased versus wild-type cells. Furthermore, the amounts of two different human proteins on BacMPs were successfully improved. This phenomenon could be due to the reduction of the degradation of target proteins in the Δlon strain. This is the first report to construct a protease deletion mutant in magnetotactic bacteria. The Δlon strain is a useful host to provide BacMPs displaying target proteins for various experimental, and ultimately, clinical applications.

AB - Bacterial magnetic particles (BacMPs) produced by the magnetotactic bacterium Magnetospirillum magneticum AMB-1, are used as magnetic supports or carriers for a variety of biomedical and environmental applications. Although protein expression systems on BacMPs have been established in previous studies, the expression efficiency was dependent on the introduced protein sequences. Recombinant human proteins are often poorly expressed on BacMPs because of proteolytic degradation by endogenous proteases. We constructed a lon protease gene deletion mutant strain (Δlon) of M. magneticum AMB-1 by homologous recombination to increase the efficiency of functional protein display on BacMPs using Δlon host cells. Wild-type and Δlon-M. magneticum AMB-1 cells were transformed using expression plasmids for human proteins, thyroid-stimulating hormone receptor (TSHR) and the class II major histocompatibility complex (MHC II) molecules onto BacMPs. Although mRNA expression of both TSHR and MHC II was the same level in the wild-type and Δlon transformants, the protein expression levels in Δlon transformants were significantly increased versus wild-type cells. Furthermore, the amounts of two different human proteins on BacMPs were successfully improved. This phenomenon could be due to the reduction of the degradation of target proteins in the Δlon strain. This is the first report to construct a protease deletion mutant in magnetotactic bacteria. The Δlon strain is a useful host to provide BacMPs displaying target proteins for various experimental, and ultimately, clinical applications.

KW - Bacterial magnetic particles

KW - Deletion mutant

KW - Lon protease

KW - Magnetotactic bacteria

KW - Protein display

UR - http://www.scopus.com/inward/record.url?scp=84878202662&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84878202662&partnerID=8YFLogxK

U2 - 10.1016/j.jbiosc.2013.01.016

DO - 10.1016/j.jbiosc.2013.01.016

M3 - Article

C2 - 23578586

AN - SCOPUS:84878202662

VL - 116

SP - 65

EP - 70

JO - Journal of Bioscience and Bioengineering

JF - Journal of Bioscience and Bioengineering

SN - 1389-1723

IS - 1

ER -