Production of luciferase-magnetic particle complex by recombinant Magnetospirillum sp. AMB-1

Tadashi Matsunaga, H. Togo, T. Kikuchi, T. Tanaka

Research output: Contribution to journalArticle

42 Citations (Scopus)

Abstract

Luciferase-bacterial magnetic particle (BMP) complexes were produced by recombinant Magnetospirillum sp. AMB-1. We constructed plasmids pKML and pNELM, respectively, by fusing luc to the 5' and 3' terminal of magA, encoding an integral iron translocating protein situated in the BMP membrane, of AMB-1. In addition, we produced bifunctional active-fusion proteins on BMPs by using a plasmid pAcML. In this plasmid, acetate kinase and luciferase genes were fused to the N-terminus and the C-terminus of MagA, respectively. Bacterial magnetic particles isolated from transconjugants for pKML, pNELM and pAcML exhibited luciferase activity. Bacterial magnetic particles isolated from transconjugants for pAcML also exhibited acetate kinase activity. Fed-batch culture of pKML transconjugant yielded 2.6 mg BMPs per liter of culture, and 95% conversion of iron into magnetite was obtained, at a nitrate concentration of 1.4 mM. Continuous feeding of iron as ferric quinate significantly enhanced growth and total magnetic production. Final cell concentration of 1.8 x 109 cells/mL and 6 mg per liter of culture was obtained. Magnetite production by fed-batch culture of AMB-1 was about 3 times that obtained by batch culture. There were no significant differences in BMPs yield between recombinant AMB-1 cultivated by fed-batch culture and wild type of AMB-1. (C) 2000 John Wiley and Sons, Inc.

Original languageEnglish
Pages (from-to)704-709
Number of pages6
JournalBiotechnology and Bioengineering
Volume70
Issue number6
DOIs
Publication statusPublished - 2000 Dec 20
Externally publishedYes

Keywords

  • Bacterial magnetic particle (BMP)
  • BMP-luciferase complex
  • Fed-batch culture
  • MagA
  • Magnetospirillum sp. AMB-1

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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