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 language | English |
|---|---|
| Pages (from-to) | 704-709 |
| Number of pages | 6 |
| Journal | Biotechnology and Bioengineering |
| Volume | 70 |
| Issue number | 6 |
| DOIs | |
| Publication status | Published - 2000 Dec 20 |
| Externally published | Yes |
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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
Cite this
Production of luciferase-magnetic particle complex by recombinant Magnetospirillum sp. AMB-1. / Matsunaga, Tadashi; Togo, H.; Kikuchi, T.; Tanaka, T.
In: Biotechnology and Bioengineering, Vol. 70, No. 6, 20.12.2000, p. 704-709.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Production of luciferase-magnetic particle complex by recombinant Magnetospirillum sp. AMB-1
AU - Matsunaga, Tadashi
AU - Togo, H.
AU - Kikuchi, T.
AU - Tanaka, T.
PY - 2000/12/20
Y1 - 2000/12/20
N2 - 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.
AB - 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.
KW - Bacterial magnetic particle (BMP)
KW - BMP-luciferase complex
KW - Fed-batch culture
KW - MagA
KW - Magnetospirillum sp. AMB-1
UR - http://www.scopus.com/inward/record.url?scp=0034694884&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0034694884&partnerID=8YFLogxK
U2 - 10.1002/1097-0290(20001220)70:6<704::AID-BIT14>3.0.CO;2-E
DO - 10.1002/1097-0290(20001220)70:6<704::AID-BIT14>3.0.CO;2-E
M3 - Article
C2 - 11064341
AN - SCOPUS:0034694884
VL - 70
SP - 704
EP - 709
JO - Biotechnology and Bioengineering
JF - Biotechnology and Bioengineering
SN - 0006-3592
IS - 6
ER -