Increasing in vivo iron loading of archaeoglobus fulgidus ferritin (afftn) transformed E.coli by overexpressing the ferrous transporter, FeoB
Research has proven iron-loaded Archaeoglobus fulgidus ferritin (AfFtn) as a good T2 MRI contrast agent and that higher concentrations of iron in iron-loaded AfFtn enhanced T2 imaging contrasts. Works on increasing in vivo AfFtn iron loading using E. coli as a biofactory have been performed by incre...
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sg-ntu-dr.10356-452822023-03-03T15:32:12Z Increasing in vivo iron loading of archaeoglobus fulgidus ferritin (afftn) transformed E.coli by overexpressing the ferrous transporter, FeoB Tan, Ker Jia. Lim Sierin School of Chemical and Biomedical Engineering DRNTU::Engineering::Chemical engineering::Biotechnology Research has proven iron-loaded Archaeoglobus fulgidus ferritin (AfFtn) as a good T2 MRI contrast agent and that higher concentrations of iron in iron-loaded AfFtn enhanced T2 imaging contrasts. Works on increasing in vivo AfFtn iron loading using E. coli as a biofactory have been performed by increasing the extracellular ferrous (Fe2+) concentrations. However, the amount of iron loaded achieved at 1400 Fe/24-mer is still far from the ~7200 Fe/24-mer stoichiometric value. It was hypothesized that the E.coli cytoplasmic membrane is the limiting factor to high intracellular Fe2+ concentrations. This project investigated the possibility of increasing ferrous iron permeability of the E.coli cytoplasmic membrane by overexpressing its ferrous transporter, FeoB, in order to increase the intracellular Fe2+ concentration and subsequently increasing in vivo iron loading into cytoplasmic AfFtn. AfFtn and FeoB from BL21 (DE3) were co-expressed in E.coli BL21 (DE3) CodonPlus-RIL cells. FeoB was tagged with green fluorescent protein (GFP) to detect its correct expression. Iron loading was compared across FeoB-GFP-AfFtn, FeoB-AfFtn and AfFtn transformed E.coli at 2 and 4 hours of 10mM Fe2+ iron loading. Results obtained shown that a ~3 fold increase in AfFtn iron loading was achieved when FeoB-GFP was co-expressed with AfFtn. Thus, overexpression of ferrous transporter FeoB increased the efficiency of in vivo iron nanoparticles production using E.coli as a biofactory. Bachelor of Engineering (Chemical and Biomolecular Engineering) 2011-06-10T07:07:30Z 2011-06-10T07:07:30Z 2011 2011 Final Year Project (FYP) http://hdl.handle.net/10356/45282 en Nanyang Technological University 66 p. application/pdf |
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DRNTU::Engineering::Chemical engineering::Biotechnology Tan, Ker Jia. Increasing in vivo iron loading of archaeoglobus fulgidus ferritin (afftn) transformed E.coli by overexpressing the ferrous transporter, FeoB |
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Research has proven iron-loaded Archaeoglobus fulgidus ferritin (AfFtn) as a good T2 MRI contrast agent and that higher concentrations of iron in iron-loaded AfFtn enhanced T2 imaging contrasts. Works on increasing in vivo AfFtn iron loading using E. coli as a biofactory have been performed by increasing the extracellular ferrous (Fe2+) concentrations. However, the amount of iron loaded achieved at 1400 Fe/24-mer is still far from the ~7200 Fe/24-mer stoichiometric value. It was hypothesized that the E.coli cytoplasmic membrane is the limiting factor to high intracellular Fe2+ concentrations. This project investigated the possibility of increasing ferrous iron permeability of the E.coli cytoplasmic membrane by overexpressing its ferrous transporter, FeoB, in order to increase the intracellular Fe2+ concentration and subsequently increasing in vivo iron loading into cytoplasmic AfFtn. AfFtn and FeoB from BL21 (DE3) were co-expressed in E.coli BL21 (DE3) CodonPlus-RIL cells. FeoB was tagged with green fluorescent protein (GFP) to detect its correct expression. Iron loading was compared across FeoB-GFP-AfFtn, FeoB-AfFtn and AfFtn transformed E.coli at 2 and 4 hours of 10mM Fe2+ iron loading. Results obtained shown that a ~3 fold increase in AfFtn iron loading was achieved when FeoB-GFP was co-expressed with AfFtn. Thus, overexpression of ferrous transporter FeoB increased the efficiency of in vivo iron nanoparticles production using E.coli as a biofactory. |
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Lim Sierin |
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Lim Sierin Tan, Ker Jia. |
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Final Year Project |
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Tan, Ker Jia. |
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Tan, Ker Jia. |
title |
Increasing in vivo iron loading of archaeoglobus fulgidus ferritin (afftn) transformed E.coli by overexpressing the ferrous transporter, FeoB |
title_short |
Increasing in vivo iron loading of archaeoglobus fulgidus ferritin (afftn) transformed E.coli by overexpressing the ferrous transporter, FeoB |
title_full |
Increasing in vivo iron loading of archaeoglobus fulgidus ferritin (afftn) transformed E.coli by overexpressing the ferrous transporter, FeoB |
title_fullStr |
Increasing in vivo iron loading of archaeoglobus fulgidus ferritin (afftn) transformed E.coli by overexpressing the ferrous transporter, FeoB |
title_full_unstemmed |
Increasing in vivo iron loading of archaeoglobus fulgidus ferritin (afftn) transformed E.coli by overexpressing the ferrous transporter, FeoB |
title_sort |
increasing in vivo iron loading of archaeoglobus fulgidus ferritin (afftn) transformed e.coli by overexpressing the ferrous transporter, feob |
publishDate |
2011 |
url |
http://hdl.handle.net/10356/45282 |
_version_ |
1759853176744312832 |