• Journal of Microbiology and Biotechnology
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• v.14 no.5
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• pp.1043-1046
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• 2004

The stability and electrochemical properties of a self-assembled layer of spinach ferredoxin on a quartz substrate and on a highly oriented pyrolytic graphite electrode were investigated. To fabricate the ferredoxin self-assembly layer, dimyristoylphosphatidylcholine was first deposited onto a substrate for ferredoxin immobilization. Surface analysis of the ferredoxin layer was carried out by atomic force microscopy to verify the ferredoxin immobilization. To verify ferredoxin immobilization on the lipid layer and to confirm the maintenance of redox activity, absorption spectrum measurement was carried out. Finally, cyclic-voltammetry measurements were performed on the ferredoxin layers and the redox potentials were obtained. The redox potential of immobilized ferredoxin had a formal potential value of -540 mV. It is suggested that the redox-potential measurement of self-assembled ferredoxin molecules could be used to construct a biosensor and biodevice.

• 한국생물공학회:학술대회논문집
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• 2003.10a
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• pp.209-213
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• 2003

Self-assembled monolayers of spinach ferredoxin immobilized to Au substrate were investigated. Ferredoxin was immobilized onto the chemically modified Au surface. Au surface were modified to $NH^{3+}$ by the 4-aminothiphenol and then modified by N-succinimidyl-3-[2-pyridyldithio]propionate for the ferredoxin immobilization. To verify the electrochemical activity of immobilized ferredoxin molecules, cyclic-voltammetry was measured. Finally, to verify the memory application, reduction potential was applied to ferredoxin molecules as for the write function, and then current transients observed from the reduced ferredoxin layers were measured for the read function of memory applications.

• Journal of Microbiology and Biotechnology
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• v.16 no.1
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• pp.15-19
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• 2006

A ferredoxin adsorbed hetero self-assembled layer was fabricated on chemically modified Au substrate, 4-Aminothiophenol (4-ATP) was deposited onto Au substrate and then N-succinimidyl-3-[2-pyridyldithio] propionate (SPDP) was adsorbed on the 4-ATP layer, since SPDP was used as a bridging molecule for ferredoxin adsorption, Ferredoxin/SPDP/4-ATP structured hetero layer was constructed because of strong chemical binding of ferredoxin, SPDP, and 4-ATP, The surface of the ferredoxin-adsorbed SPDP/4-ATP layer was observed by scanning tunneling microscopy, The hetero film formation was verified by surface plasmon resonance measurement. The current flow and rectifying property based on the scanning tunneling spectroscopy I-V characteristics was achieved in the proposed hetero layer. Thus, the hetero layer structure of ferredoxin functioned as a molecular diode with rectifying property, The proposed molecular diode can be usefully applied for the development of molecular scale electronic devices.

• BMB Reports
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• v.39 no.1
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• pp.46-54
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• 2006

The coenzyme A-acylating 2-oxoacid:ferredoxin oxidoreductase and ferredoxin (an effective electron acceptor) were purified from the hyperthermophilic archaeon, Sulfolobus solfataricus P1 (DSM1616). The purified ferredoxin is a monomeric protein with an apparent molecular mass of approximately 11 kDa by SDS-PAGE and of $11,180{\pm}50$ Da by MALDI-TOF mass spectrometry. Ferredoxin was identified to be a dicluster, [3Fe-4S][4Fe-4S], type ferredoxin by spectrophotometric and EPR studies, and appeared to be zinc-containing based on the shared homology of its N-terminal sequence with those of known zinc-containing ferredoxins. On the other hand, the purified 2-oxoacid: ferredoxin oxidoreductase was found to be a heterodimeric enzyme consisting of 69 kDa $\alpha$ and 34 kDa $\beta$ subunits by SDS-PAGE and MALDI-TOF mass spectrometry. The purified enzyme showed a specific activity of 52.6 units/mg for the reduction of cytochrome c with 2-oxoglutarate as substrate at $55^{\circ}C$, pH 7.0. Maximum activity was observed at $70^{\circ}C$ and the optimum pH for enzymatic activity was 7.0 -8.0. The enzyme displays broad substrate specificity toward 2-oxoacids, such as pyruvate, 2-oxobutyrate, and 2-oxoglutarate. Among the 2-oxoacids tested (pyruvate, 2-oxobutyrate, and 2-oxoglutarate), 2-oxoglutarate was found to be the best substrate with $K_m$ and $k_{cat}$ values of $163\;{\mu}M$ and $452\;min^{-1}$, respectively. These results provide useful information for structural studies on these two proteins and for studies on the mechanism of electron transfer between the two.

• Journal of Microbiology and Biotechnology
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• v.21 no.2
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• pp.124-128
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• 2011

Ferredoxin is a typical iron-sulfur protein that is ubiquitous in biological redox systems. This study investigates the in vitro assembly of a [$Fe_2S_2$] cluster in the ferredoxin from Acidithiobacillus ferrooxidans in the presence of three scaffold proteins: IscA, IscS, and IscU. The spectra and MALDI-TOF MS results for the reconstituted ferredoxin confirm that the iron-sulfur cluster was correctly assembled in the protein. The inactivation of cysteine desulfurase by L-allylglycine completely blocked any [$Fe_2S_2$] cluster assembly in the ferredoxin in E. coli, confirming that cysteine desulfurase is an essential component for iron-sulfur cluster assembly. The present results also provide strong evidence that [$Fe_2S_2$] cluster assembly in ferredoxin follows the AUS pathway.

• Microbiology and Biotechnology Letters
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• v.25 no.4
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• pp.367-373
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• 1997

One of the three components of the phenanthrene dioxygenase which is required for conversion of phenanthrene to cis-phenanthrene dihydrodiol, Rieske-type ferredoxin encoded by phnR has been cloned and sequenced from Pseudomonas sp. strain DJ77. The gene phnR is positioned at the downstream of phnQ encoding 2,3-dihydroxybiphenyl 1,2-dioxygenase. The PhnR ferredoxin contains 108 amino acids with a Mr of 11,355. The deduced amino acid sequence of the PhnR ferredoxin is 35-79% identical to those of homologous ferredoxins encoded by various genes.

• Korean Journal of Microbiology
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• v.34 no.3
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• pp.115-119
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• 1998

We cloned the 4.8 kb BglII fragment containing genes downstream pHENX7 from Pseudomonas sp. strain DJ77. The restriction map of the resultant clone, recombinant plasmid pYCS500, was determined. Sequencing analysis of the 465 bp HindIII-ClaI fragment revealed an open reading frame of 282 bp that was then designated phnM. The deduced polypeptide is 93 amino acid residues long with a $M_r$ of 10,008. The PhnM has 37.3-53.9% identity with plant-type ferredoxin proteins such as NahT, XylT, DmpQ, AtdS, PhlG, PhhQ and TbuW and contains the motif similar to well-conserved functional domains of those proteins.

• BMB Reports
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• v.35 no.4
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• pp.432-436
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• 2002

Pseudomonas sp. S-47 is capable of degrading catechol and 4-chlorocatechol via the meta-cleavage pathway. XyITE products catalyze the dioxygenation of the aromatics. The sylT of the strain S-47 is located just upstream of the xylE gene. XylT of the strain S-47 is located just upstream of the xylE gene. XyIT is typical chloroplast-type ferredoxin, which is characterized by 4 cystein residues that are located at positions 41, 46, 49, and 81. The chloroplast-type ferredoxin of Pseudomonas sp. S-47 exhibited a 98% identity with that of P. putida mt-2(TOL plasmid) in the amino acid sequence, but only about a 40 to 60% identity with the corresponding enzymes from other organisms. We constructed two recombinant plasmids (pRES1 containing xylTE and pRES101 containing xylE without xylT) in order to examine the function of XyIT for the reactivation of the catechol 2,3-dioxygenase (XyIE) that is oxidized with hydrogen peroxide was recovered in the catechol 2,3-dioxygenase (C23O) activity about 4 mimutes after incubation, but the pRES101 showed no recovery. That means that the typical chloroplast-type ferredoxin (XyIT) of Pseudomonas sp. S-47 is involved in the reactivation of the oxidized C23O in the dioxygenolytic cleavage of aromatic compounds.

• Parasites, Hosts and Diseases
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• v.54 no.1
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• pp.71-74
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• 2016

Trichomonas vaginalis is a flagellate protozoan parasite and commonly infected the lower genital tract in women and men. Iron is a known nutrient for growth of various pathogens, and also reported to be involved in establishment of trichomoniasis. However, the exact mechanism was not clarified. In this study, the author investigated whether the 120 kDa protein of T. vaginalis may be involved in pathogenicity of trichomonads. Antibodies against 120 kDa protein of T. vaginalis, which was identified as pyruvate:ferredoxin oxidoreductase (PFOR) by peptide analysis of MALDI-TOF-MS, were prepared in rabbits. Pretreatment of T. vaginalis with anti-120 kDa Ab decreased the proliferation and adherence to vaginal epithelial cells (MS74) of T. vaginalis. Subcutaneous tissue abscess in anti-120 kDa Ab-treated T. vaginalis-injected mice was smaller in size than that of untreated T. vaginalis-infected mice. Collectively, the 120 kDa protein expressed by iron may be involved in proliferation, adhesion to host cells, and abscess formation, thereby may influence on the pathogenicity of T. vaginalis.

• Journal of Microbiology
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• v.34 no.2
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• pp.137-143
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• 1996

We isolated a promoter inducible by paraquat, a superoxide-generating agent, from Escherichia coli using a promoter-probing plasmid pRS415. From sequence analysis we found out the promoter is for fpr ENCODING nadph : ferredoxin oxidoreductase. We constructed on operon fusion of lacZ gene with fpr promoter to monitor the expression of the gene in the single-copy state. LacZ expression generators, menadione and plumbagin, also induced the expression of .betha.-galactosidase in the fusion strain. On the other hand, no significant induction was observed by treatment with hydrogen peroxide, ethanol, and heat shock. Induction of .betha.-galactosidase was significantly reduced by introducing a .DELTA. sox 8 :: cat of soxS3 :: Tn10 mutation into the fusion strain, indicating that fpr gene is a member of the soxRS regulon. The transcriptional start site was determined by primer extension analysis. Possible roles of fpr induction in superoxide stress were discussed.