Development of a peroxidase-Fused protein reagent by brevibacillus choshinensis heterologous expression system

Kounosuke Hayashi, Yusuke Tomozoe, Kenji Nagai, Yoshiyuki Hiraishi, Noriho Kamiya

Research output: Contribution to journalArticle

Abstract

To detect a target protein in biological samples, a fusion protein was designed composed of a peroxidase from Arthromyces ramosus (ARP) and parts of the antibody-binding domains of Staphylococcus aureus protein A and Streptococcus protein G (PG). The ARP-PG fusion protein was successfully expressed by a heterologous protein expression system in Brevibacillus choshinensis. The fusion protein was secreted as an active form in culture media. The production of ARP-PG with higher peroxidase activity was observed by the addition of 5-aminolevulinic acid to the culture media. The performance of purified ARP-PG was validated by dot blotting for the detection of transferrin as a model target protein. A comparable performance in the dot blot analysis was attained using a culture supernatant containing crude but active ARP-PG, indicating the practicality of the Brevibacillus protein secretion system.

Original languageEnglish
Pages (from-to)157-161
Number of pages5
Journalkagaku kogaku ronbunshu
Volume41
Issue number2
DOIs
Publication statusPublished - Mar 20 2015

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Peroxidase
Proteins
Fusion reactions
Culture Media
Aminolevulinic Acid
Staphylococcal Protein A
Transferrin
Antibodies

All Science Journal Classification (ASJC) codes

  • Chemistry(all)
  • Chemical Engineering(all)

Cite this

Development of a peroxidase-Fused protein reagent by brevibacillus choshinensis heterologous expression system. / Hayashi, Kounosuke; Tomozoe, Yusuke; Nagai, Kenji; Hiraishi, Yoshiyuki; Kamiya, Noriho.

In: kagaku kogaku ronbunshu, Vol. 41, No. 2, 20.03.2015, p. 157-161.

Research output: Contribution to journalArticle

Hayashi, Kounosuke ; Tomozoe, Yusuke ; Nagai, Kenji ; Hiraishi, Yoshiyuki ; Kamiya, Noriho. / Development of a peroxidase-Fused protein reagent by brevibacillus choshinensis heterologous expression system. In: kagaku kogaku ronbunshu. 2015 ; Vol. 41, No. 2. pp. 157-161.
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