• Journal of Periodontal and Implant Science
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• v.29 no.3
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• pp.663-676
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• 1999

Porphyromonas gingivalis, a Gram negative, anaerobic, asaccharolytic rod, is one of the most frequently implicated pathogens in human periodontal disease and has a requirement for hemin for growth. A 30 kDa (heated 24 kDa) hemin-binding protein whose expression is both hemin and iron regulated has recently been purified and characterized in this oral pathogen. This study has identified a hemin-binding P. gingivalis protein by expression of a P. gingivalis genomic library in Escherichia coli, a bacterium which does not require or transport exogenous hemin. A library of genomic DNA fragments from P. gingivalis was constructed in plasmid pUC18, transformed into Escherichia coli strain $DH5{\alpha}$ , and screened for recombinant clones with hemin-binding activity by plating onto hemin-containing agar. Of approximately 10,000 recombinant E. coli colonies screened on LB-amp-hemin agar, 10 exhibited a clearly pigmented phenotype. Each clone contained various insert DNA. The Hind III fragment transferred to the T7 RNA polymerase/promoter expression vector system produced a sligltly smaller (21 kDa) protein, a precursor form, immunoreactive to the antibody against the 24 kDa protein, suggesting that the cloned DNA fragment probably carried an entire gene for the 24 kDa hemin-binding protein.

• Journal of Periodontal and Implant Science
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• v.36 no.1
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• pp.155-165
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• 2006

The results of this study confirm that the availability of hemin influences the expression of selected membrane proteins of Porphyromonas gingivalis, Prevotella intermedia, and Prevotella nigrescens. A 30 kDa (heated 24 kDa) hemin-binding protein whose expression is hemin regulated was identified and purified in P. gingivalis. A strong hemin-binding function was found by LDS-PAGE and TMBZ staining when P. gingivalis cells were grown under hemin-limited conditions. A 50 kDa cell envelope associated protein, whose expression is hemin regulated, is considered to be a putative hemin binding protein from P. intermedia and P. nigrescens, respectively. N-terminal amino acid sequence analysis of CNBr-digested 24 kDa hemin binding protein from P. gingivalis revealed that this protein belongs to a new, so far undescribed hemin-binding class of proteins. N-terminal amino acid sequence of a 50 kDa putative hemin binding protein from P. intermedia was identical with Enolase from Streptococcus intermedia. Work is in progress to further characterize the molecular structure of these proteins.

• Analytical Science and Technology
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• v.7 no.4
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• pp.505-515
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• 1994

$^1H$ NMR spectra for monocyanide ligated ferriprotoporphyrin(hemin) complex and dicyanide coordinated hemin complex in dimethylsulfoxide(DMSO-$d_6$) solution have been recorded and analyzed. NMR spectra of hemin-cyanide complexation in DMSO-$d_6$ exhibit that the cyanide ligation to hemin is temperature-dependent. Thermodynamic parameters for the monocyanide ligated hemin to dicyanide ligated hemin are consistent with endothermic process with ${\Delta}H^{\circ}=736.6cal/mol$ and ${\Delta}S^{\circ}=16.4eu$. Detailed analysis of the anomalous deviation from Curie behavior for CN/DMSO coordinated hemin complex demonstrates the presence of a high spin character, and this weaker axial field relative to the purely low-spin dicyanide hemin complex is supposed to attribute to instantaneously ruptured iron-DMSO bond. This complex may serve as a useful model to characterize electronic/molecular structure of hemoproteins, which one of axial ligands is weak.

• Journal of Periodontal and Implant Science
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• v.30 no.4
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• pp.737-746
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• 2000

본 연구는 hemin이 치주질환 주요 병인균주 중의 하나인 Prevotella intermedia의 성장 및 세포막 단백질의 발현에 미치는 영향을 규명하고, hemin 결합에 관여하는 것으로 추정되는 단백질의 순수분리 및 특성 분석을 위해 수행 되었다. 본 연구의 결과에 의하면, hemin은 P.intermedia의 성장 및 세포막 단백질의 발현에 영향을 미쳐, hemin이 고갈된 조건에서 균주의 성장이 현저히 억제되었으며, 약 50 kDa의 세포막 단백질이 현저히 강화되어 발현되었다. 본 연구에서는 hemin 결합에 관여하는 것으로 추정되는 이 50 kDa의 세포막 단백질을 순수분리하였으며, N'-terminal 아미노산 분석을 수행한 결과 이 단백질은 Streptococcus inter - medius의 Enolase와 아미노산 서열 및 분자량이 일치하였다. 한편, 이 단백질에는 disulfidebond가 존재하지 않았다. 본 연구는 P.intermedia에서의 porphyrin 생리 및 hemin 획득기전을 밝히는데 있어 중요한 의의가 있으리라 사료된다.

• Korean Chemical Engineering Research
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• v.60 no.4
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• pp.588-593
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• 2022

Terephthalaldehyde (TPA) is introduced as a cross liker to enhance electron transfer of hemin-based cathodic catalyst consisting of polyethyleneimine (PEI), carbon nanotube (CNT) for hydrogen peroxide reduction reaction (HPRR). In the cyclic voltammetry (CV) test with 10 mM H₂O₂ in phosphate buffer solution (pH 7.4), the current density for HPRR of the suggested catalyst (CNT/PEI/hemin/PEI/TPA) shows 0.2813 mA cm^-2 (at 0.2 V vs. Ag/AgCl), which is 2.43 and 1.87 times of non-cross-linked (CNT/PEI/hemin/PEI) and conventional cross liker (glutaraldehyde, GA) used catalyst (CNT/PEI/hemin/PEI/GA), respectively. In the case of onset potential for HPRR, that of CNT/PEI/hemin/PEI/TPA is observed at 0.544 V, while those of CNT/PEI/hemin/PEI and CNT/PEI/hemin/PEI/GA are 0.511 and 0.471 V, respectively. These results indicate that TPA plays a role in facilitating electron transfer between the electrodes and substrates due to the π-conjugated cross-linking bonds, whereas conventional GA cross-linker increases the overpotential by interrupting electron and mass transfer. Electrochemical impedance spectroscopy (EIS) results also display the same tendency. The charge transfer resistance (R_ct) of CNT/PEI/hemin/PEI/TPA decreases about 6.2% from that of CNT/PEI/hemin/PEI, while CNT/PEI/hemin/PEI/GA shows the highest R_ct. The polarization curve using each catalyst also supports the superiority of TPA cross liker. The maximum power density of CNT/PEI/hemin/PEI/TPA (36.34±1.41 μWcm^-2) is significantly higher than those of CNT/PEI/hemin/PEI (27.87±0.95 μWcm^-2) and CNT/PEI/hemin/PEI/GA (25.57±1.32 μWcm^-2), demonstrating again that the cathode using TPA has the best performance in HPRR.

• Animal Bioscience
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• v.34 no.8
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• pp.1382-1391
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• 2021

Objective: The objective was to evaluate the impact of different forms of iron including myoglobin, hemin, and ferric chloride on the quality of chicken breast meat. Methods: Chicken breast muscles were subjected to 1, 2, 3 mg/mL of FeCl₃, myoglobin and hemin treatment respectively, and the production of reactive oxygen species (ROS) and malondialdehyde, meat color, tenderness, water holding capacity and morphology of meat was evaluated. Results: Hemin was found to produce more ROS and induce greater extent of lipid oxidation than myoglobin and ferric chloride. However, it showed that hemin could significantly increase the redness and decrease the lightness of the muscle. Hemin was also shown to be prominent in improving water holding capacity of meat, maintaining a relatively higher level of the immobilized water from low-field nuclear magnetic resonance measurements. Morphology observation by hematoxylin-eosin staining further confirmed the results that hemin preserved the integrity of the muscle. Conclusion: The results indicated that hemin may have economic benefit for the industry based on its advantage in improving water holding capacity and quality of meat.

• Archives of Pharmacal Research
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• v.26 no.5
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• pp.411-415
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• 2003

To investigate the mechanisms involved in hemin-induced febrile response, the rectal temperature of rats were measured after intracerebroventricular (i.c.v.) injections of hemin, with or without antagonists. Hemin ($10\mu\textrm{g}$) elicited a significant febrile response, which lasted from 30 min, to more than 6 h, after its administration, but this was not the case with biliverdin (i.c.v.) and bilirubin (i.c.v.). The hemin-induced febrile response was significantly inhibited by pretreatment with an inhibitor of tyrosine kinase (genistein), but not by pretreatment with an inhibitor of protein kinase C (chelerythrine) and a scavenger of iron (deferoxamine). These results suggest that tyrosine kinase is involved in the hemin-induced febrile response.

• Bulletin of the Korean Chemical Society
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• v.6 no.2
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• pp.112-115
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• 1985

3-Aminopropylsilated silica gel bound hemin, biliverdin, and bilirubin were synthesized by reacting 3-aminopropyl silated silica gel with hemin, biliverdin and bilirubin respectively. The aspects of binding of bilirubin to amino group of 3-aminopropylsilated silica gel were studied using the above synthetic silica immobilized hemin, biliverdin and bilirubin, and oxodipyrromethene.

• Journal of the korean academy of Pediatric Dentistry
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• v.33 no.2
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• pp.304-310
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• 2006

Prevotella intermedia is one of the most frequently implicated pathogens in human periodontal disease and has a requirement for hemin for growth. This study has identified a hemin-binding P. intermedia protein by expression of a P. intermedia genomic library in Escherichia coli, a bacterium which does not require or transport exogenous hemin. The genomic library of P. intermedia was constructed into plasmid pUC18, transformed into Escherichia coli strain $DH5{\alpha}$, and screened for recombinant clones using heminbinding activity by plating onto hemin-containing agar. Approximately 5,000 recombinant E. coli colonies were screened onto LB-amp-hemin agar, single clone(pHem1) was exhibited a clearly pigmented phonotype. The 2.5kb insert DNA of pHem1 was determined by restriction enzyme mapping. Southern blot analysis of BamHI, BglII, EcoRI, HindIII and PstI-digested P. intermedia DNA indicated that single copy of the gene was present in the genome. Northern blot analysis revealed that the size of transcript was approximately 1.8 kb. The cloned gene contained a single ORF, consisting of approximately 850-residue amino acids. A BLAST search of the Institute for Genomic Research genes with similar nucleotide sequence revealed no significant similarity It needs further investigation to clarify the mechanisms of heme uptake in P. intermedia.

• Journal of Periodontal and Implant Science
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• v.32 no.2
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• pp.351-360
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• 2002

In the study presented here, identification, purification, and partial characterization of a hemin-regulated protein in Prevotella nigrescens were carried out. The results of this study confirm that the availability of hemin influences the expression of a selected membrane protein as well as the growth rate of P. nigrescens ATCC 33563. The 50 kDa cell envelope associated protein, whose expression is hemin regulated, is considered to be a putative hemin-binding protein from P. nigrescens. Disulfide bonds were not present in this protein, and N'-terminal amino acid sequence analysis revealed that this protein belongs to a new, so far undescribed protein. The 50 kDa protein was found to be rich in hydrophilic amino acids, with glycine comprising approximately 60% of the total amino acids. The study described here is the first to identify, purify, and biochemically characterize a putative hemin-binding protein from P. nigrescens. Work is in progress to further characterize the molecular structure of this protein.