• Applied Biological Chemistry
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• 제44권4호
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• pp.219-223
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• 2001

Hemoglobin(Hgb)의 효소 가수분해액으로부터 고농도 heme-iron의 분리에 영향을 주는 기질의 농도, 가수분해도 (DH, degree of hydrolysis) 및 분리 pH의 효과 등을 조사하였으며, pilot scale에서 고농도 heme-iron을 생산하였다. 최적 반응조건(pH 10.0, $50^{\circ}C$, 5 hr)에서의 Esperase에 의한 Hgb의 DH는 $24{\sim}25%$였다. Hgb의 농도는 heme-iron의 분리에 영향을 미쳐서 Hgb의 농도가 2.5%, 5.0%, 10.0%로 증가할수록 heme-iron/peptide(heme-iron의 분리도)는 각각 47.4%, 35.6%, 21.8%로 감소하였으며, 이에 따라 농축율도 감소하였다. Hgb의 가수분해액으로부터 고농도 heme-iron의 분리에 효율적인 pH 영역은 DH에 따라 달라졌으며, DH가 6.3%의 경우는 pH $5.0{\sim}6$에서 heme-iron/peptide의 비가 8.92%, DH가 14.5%인 경우는 pH $4.0{\sim}6.0$에서 $12.1{\sim}19.5%$였으며, DH가 24.3%인 경우는 pH $3.0{\sim}5.0$에서 $34.7{\sim}36.4%$로 가수분해도가 높을수록 고농도 heme-iron의 분리시 최적 PH 범위가 산성 영역으로 이동되었으며, 이 영역의 pH에서 peptide의 용해도 증가로 분리도가 증가되었다. 가수분해액으로부터 분리된 heme-iron의 분자량은 약 1 kDa정도로 hematin과 유사한 분리패턴을 보였다. Pilot scale로 제조한 고농도 heme-iron 제품에서의 heme-iron 함량은 27.1%, heme-iron/peptide 비는 38.7%, 생산수율은 9.3%이었다.

• Applied Biological Chemistry
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• 제46권2호
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• pp.130-133
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• 2003

Hemoglobin(Hgb)의 가수분해물로부터 heme-iron을 분리하는데 영향을 미치는 Hgb의 농도, 가수분해도(DH, degree of hydrolysis), 투석용액의 pH, 투석막의 pore size에 대하여 조사하였다. 가수분해에 사용된 Hgb의 농도가 높아짐에 따라 heme-iron의 회수율은 증가하였으나, peptide의 제거율과 HP ratio(총 peptide 중 heme-iron이 차지하는 비율)는 Hgb의 농도에 따른 큰 차이를 볼 수 없었다. Hgb의 가수분해도가 8%, 16%, 24%로 증가함에 따라 투석에 의한 peptide의 제거가 용이하게 이루어져 HP ratio가 13.7%, 20.7%, 31%로 증가하였다. 투석용액에 $KH_2PO_4$를 25 mM이 되도록 첨가할 때는 HP ratio가 25.7%이었으나,50 mM이상의 농도에서는 30.0-32.5%로 증가하였다. 투석용액의 pH 높을수록 heme-iron의 용해도가 증가하여 투석막을 통한 heme-iron의 세출이 많아졌다. Peptide의 제거율은 투석막이 2kDa에서 5kDa로 커질 때 74.5%에서 87.5%의 큰 폭으로 증가하였으며, heme-iron의 회수율은 투석막의 크기에 따라 감소하여 2kDa에서는 86.5%, 15kDa에서는 79.6%, 25kDa에서 63.1%로 급감하였다. 투석법에 의해 분리, 건조된 heme-iron제품의 heme-iron과 peptide의 함량은 각각 21.7%와 77.0%이었으며, HP ratio는 28.2%, 수율은 6,5%이었다.

• 약학회지
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• 제45권1호
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• pp.78-84
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• 2001

Heme oxygenase is a rate-limiting enzyme in heme catabolism that cleaves heme to form biliverdin, iron, and carbon monoxide. Heme oxygenase-1 is expressed in many types of cells and tissues and is highly induced in response to oxidative stress. Carbon monoxide, one of the products of heme oxygenase, can stimulate soluble guanylate cyclase and dilate the vascular smooth muscle. So, the induction of heme oxygenase by lipopolysaccharide (LPS)-induced oxydative stress and the effect of the resultant carbon monoxide on aortic contractility were examined in this study. Zinc protoporphyrine IX (ZnPP), a inhibitor of heme oxygenase, elicited weak contraction of thoracic aortic ring, and this effect was more potent in aorta of LPS-treated rats than control and was blocked by methylene blue. The hyperreactivity to ZnPP in LPS-treated group was blocked by co-treatment with aminoguanidine. In the aortic ring of LPS-treated rats, ZnPP didn't change the vasoreactivity to phenylephrine or acetylcholine. ZnPP elicited hyper-tensive effect in concious rats, and pretreatment with LPS did not affect this effect. Prazosin significantly diminished the hypertensive effect of ZnPP. These results indicate that LPS induced heme oxygenase in aotra, and the resultant carbon monoxide diminished the aortic reactivity to vasoconstrictor.

• Environmental Engineering Research
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• 제19권2호
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• pp.139-143
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• 2014

Catalytic degradation of pentachlorophenol in soil by heme and hydrogen peroxide has been hypothesized to occur through nonspecific catalytic reactions similar to those involving ligninase. The present study examines the evidence for a heme catalytic mechanism for the oxidation of organic compounds. In the presence of hydrogen peroxide, heme is converted to the ferryl heme radical (Hm-$Fe^{+4{\cdot}}$), which can oxidize organic compounds, such as 5-aminosalicylic acid (5-ASA). A second 5-ASA may later be oxidized by ferryl heme (Hm-$Fe^{+4}$), which reverts to the ferric heme state (Hm-$Fe^{+3}$) to complete the cycle. We believe that this catalytic cycle is involved in the degradation of hazardous pollutants, such as polycyclic aromatic hydrocarbons (PAHs). Remediation via heme catalytic reactions of PAHs in soil from a pole yard was evaluated, and about 96% of PAHs was found to disappear within 42 days after treatment with heme and hydrogen peroxide. In addition, benzo[a]pyrene and six other PAHs were undetectable among a total of 16 PAH compounds examined. Therefore, we propose heme catalysis as a novel technology for the remediation of hazardous compounds in contaminated soil.

• Journal of Nutrition and Health
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• 제25권7호
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• pp.608-616
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• 1992

Using the hematofluiorometer normal values of the erythrocyte zinc protoporphyrin(ZPP) and ZPP/Heme ratio were measured in 312 males and 163 females aged from 6 month to 73 years old and compared with those of anemic persons. The mean$\pm$SD values of ZPP of normal Koreans were 28.5$\pm$6.4($\mu\textrm{g}$/dl) in males and 3.18$\pm$7.7 in females. the mean$\pm$SD of ZPP/Heme ratio were 49.5$\pm$12.3($\mu$mol/mol heme) in males and 62.0$\pm$15.8 in females. The difference in the mean ZPP and ZPP/Heme ratio values between male and female subjects were statistically signficant(p<0.0001) In male subjects the mean ZPP and ZPP/Heme ratio of the age groups less than 15 years old were higher than adult groups and the difference between age groups was significant(p<0.005 and p<0.0001 respecti-vely) The normal upper limit of the mean$\pm$2SD in normal male and female subjects were 41.3 and 47.2 for ZPP and 74.1 and 93.6 for ZPP/Heme ratio respectively. The mean values of ZPP and ZPP/Heme ratio measured in the anemic persons were higher than those of normal subjects and did not show any significant difference by the sex and age groups except in 6-14 years male groups. The test specificity(positivity) analyzed in the anemic persons by the cut-off values calculated from the normal data were 50.6% for ZPP and 73.0% for ZPP/Heme ratio. The correlation analysis between blood hemoglobin and erythrocyte ZPP or ZPP/Heme ratio in the total 801 normal and anemic subjects showed that there are very high logarithmic correlation between the hemoglobin levels and ZPP/Heme ratio (r=-0.8339) and high correla-tion between the hemoglobin levels and ZPP concentrations(r=-0.6372) These results suggested that the measurement of the erythrocyte ZPP and ZPP/Heme ratio with the hematofluorometer can be a usuful screening method for iron deficiency anermin because they provide a reliable immediate results with a small amount of sample and are relatively simple and inexpensive.

• 분석과학
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• 제11권1호
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• pp.73-78
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• 1998

미오글로빈 시안착물(MbCN) 단백질에 대한 NMR의 HMQC 연구는 수소와 결합된 상자기성 heme 탄소 시그날의 완전한 지정을 가능토록 해준다. 이러한 상자기성 MbCN에 대한 HMQC 실험의 적용은 heme시그날뿐만 아니라 상자기성 아미노산에 대해 결합된 수소와 탄소간의 coherence를 지정하여주며 자연존재량 $^{13}C$시그날의 지정이 모든 low-spin 상자기성 heme단백질에서도 가능하다. 이러한 시그날 지정 전략은 정자기성 영역에서 공명하는 수소 시그날의 지정을 위해 사용되는 NOE에만 의존하는것 보다 훨씬 명확한 시그날지정이 가능하다. 2,4-비닐기의 ${\alpha}$-탄소들과 7-프로피온기의 ${\beta}$-탄소에서 특이한 anti-Curie형태를 보이는 것은 그들이 heme평면에 존재하고 있지 않다는 증거가 된다. Proximal His에 의해 유도된 heme의 전자 및 자성의 비대칭은 heme탄소 시그날공명이 $25^{\circ}C$에서 250 ppm의 범위에 이르도록 한다. 이러한 heme 탄소 시그날 공명은 미오그로빈 heme의 전자구조를 분석하는데 있어서 수소 시그날의 공명보다 더욱 민감한 증거로 작용할수 있다.

• Journal of Microbiology and Biotechnology
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• 제19권6호
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• pp.604-609
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• 2009

To investigate the potential use of microbial heme as an iron source, recombinant Escherichia coli coexpressing ALA synthase (HemA) as well as the NADP-dependent malic enzyme (MaeB) and dicarboxylic acid transporter (DctA) were cultured. The typical red pigment extracted from the recombinant E. coli after 38 h showed highest absorbance at 407 nm, and the amount of iron in 38.4 mg of microbial heme extract derived from 6-1 fermentation broth was 4.1 mg. To determine the commercial potential of the recombinant E.coli-synthesized iron-associated heme as an iron source, mice were fed the iron-free provender with the microbial heme extract. The average body weight reduction of mice fed non-iron provender was 2.3%, whereas no detectable weight loss was evident in mice fed microbial heme addition after 15 days. The heme content of the blood from microbial heme fed mice was 4.2 mg/ml whereas that of controls was 2.4 mg/ml, which implies that the microbial heme could be available for use as an animal iron source.

• Journal of Microbiology and Biotechnology
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• 제25권6호
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• pp.880-886
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• 2015

Bacterial heme was produced from a genetic-engineered Escherichia coli via the porphyrin pathway and it was useful as an iron resource for animal feed. The amount of the E. coli-synthesized heme, however, was only few milligrams in a culture broth and it was not enough for industrial applications. To analyze heme biosynthetic pathways, an engineered E. coli artificially overexpressing ALA synthase (hemA from Rhodobacter sphaeroides) and pantothenate kinase (coaA gene from self geneome) was constructed as a bacterial heme-producing strain, and both the transcription levels of pathway genes and the intermediates concentrations were determined from batch and continuous cultures. Transcription levels of the pathway genes were not significantly changed among the tested conditions. Intracellular intermediate concentrations indicated that aminolevulinic acid (ALA) and coenzyme A (CoA) were enhanced by the hemA-coaA co-expression. Intracellular coproporphyrinogen I and protoporphyrin IX accumulation suggested that the bottleneck steps in the heme biosynthetic pathway could be the spontaneous conversion of HMB to coproporphyrinogen I and the limited conversion of protoporphyrin IX to heme, respectively. A strategy to increase the conversion of ALA to heme is discussed based on the results.

• Bulletin of the Korean Chemical Society
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• 제17권9호
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• pp.820-826
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• 1996

The p2H dependence of the NMR chemical shifts of the proton signals of heme methyl groups and ionizable groups in the vicinity of the heme were investigated. The p2H titration of heme methyl signals in four macroscopic oxidation states by saturation tranfer method was performed in the range between p2H 5.2 and 9.0. While the p2H dependence of the heme methyl resonance in fully oxidized state was small, most resonances in the intermediate oxidation states showed certain shifts. Particularly, methyl resonances of heme 1 (sequential heme numbering) exhibited sharp p2H dependence in acidic range. β-CH2 of the propionate of hemes 1 and 4 were titrated in the range of p2H 4.5-9.0. Only the 6-propionate group of heme 1 was protonated in this p2H range and its titration curve was similar to those of methyl resonances of heme 1 in intermediate oxidation states. Analysis of the microscopic redox potentials showed that they change depending on p2H. The ionizable groups responsible for the p2H dependence of these potentials are 6-propionate of heme 1 in acidic range and His 67 in basic range.

• 한국수산과학회지
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• 제35권2호
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• pp.162-165
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• 2002

백색육어를 배소한 경우와 배소한 시료를 저온에 저장 후, 단계적으로 가열처리했을 때 일어나는 산화에 의한 heme 화합물의 변화를 조사하였다. Heme 화합물 중 myoglobin 함량은 각시가자미나 조기 모두 180$^{\circ}C$에서 20분간 어피 제거육의 상태로 배소한 것이 상대적으로 감소폭이 컸으며 가열, 재가열에 의해서도 감소하는 경향이었다. Myoglobin의 감소 추세에 따라 metmyoglobin 함량은 역으로 점차 증가하는 경향이었으며, 배소온도가 낮을수록 또한 어피를 제거함으로써 생성된 metmyoglobin의 함량이 높았다. 총철 함량은 배소온도와 시간을 달리하여 처리하여도 두 어종 모두 생시료와는 함량 차이가 거의 없었다. 비 heme 철은 각시가자미의 경우 생시료에는 총철함량에 대해 약 $20\%$ 정도 함유되어 있었으나, 180$^{\circ}C$에서 20분간 어피 제거육의 상태로 배소했을 경우 생시료에 비해 약 2배 정도 증가하였다. 조기는 생시료에는 건물당으로 약 0.8mg/100g 함유되어 있었으나 배소, 가열 및 재가열과 같은 반복되는 가열과정에 의해 증가하였다. Heme 철 함량은 각시가자미와 조기 모두 비 heme 철의 증가에 반비례해서 감소하였으며 또한 배소시간이 길수록 그리고 어피 잔존육보다는 어피 제거육의 감소폭이 더 컸다.