• Applied Biological Chemistry
• /
• 제44권4호
• /
• pp.219-223
• /
• 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
• /
• 제46권2호
• /
• pp.130-133
• /
• 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.

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

• Nutritional Sciences
• /
• 제9권4호
• /
• pp.295-300
• /
• 2006

Iron bound to heme appears to be more bioavailable than iron salts. A clinical study was performed to investigate the absorption efficiency of heme-iron and iron-salt products available. Heme-iron and nonheme-iron supplements have become available in Korea. We performed iron absorption studies to compare the absorption of heme-iron polypeptide (HIP) products made from digested hemoglobin, produced in Korea (HIPk) and imported from Japan (HIj), with that of iron salts. In the study, 80 subjects were divided into 5 groups (n=56): placebo group; 12 mg glucose, HIPk group; 12 mg iron as HIPk, HIj group; 12 mg iron as HIj, iron-salt group 1; 12 mg of iron as ferrous aminoacetate, and iron-salt group 2; 100 mg iron as ferrous aminoacetate. Changes in serum iron levels were measured at 3 and 5 hours post ingestion. Absorption of iron in HIPk was higher compared to HIj, iron-salt or placebo. There was a significant inverse correlation between low serum iron levels $(<80{\mu}g/dl)$ and iron absorption from HIPk. These results demonstrated that HIPk was more bioavailable, even taken with a meal, and would have potential advantages over iron salt or HIj as an iron supplement. Our results indicate that heme-iron absorption is regulated by iron status through a heme receptor, whereas iron-salt absorption is unregulated.

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

• 한국식품영양과학회지
• /
• 제40권12호
• /
• pp.1720-1725
• /
• 2011

본 연구에서는 아스파르트산, 탄산칼슘 및 황산철을 반응시켜 아스파르트산 킬레이트 철분(aspartic acid chelated iron, Asp-Fe)을 제조하고 Asp-Fe의 철분결핍 쥐(iron-deficient rat, ID)에서의 생물학적 유용성을 확인하였다. 시험군은 철분이 함유된 식이를 섭취한 정상군(NC), 철분 결핍식이를 1개월간 투여하여 철분 결핍 상태를 유도한 쥐(ID)에 생리식염수를 공급한 결핍 대조군(ID+C), 철분 결핍 쥐에 햄철(heme-Fe) 투여군(ID+heme-Fe) 및 Asp-Fe 투여군(ID+Asp-Fe)으로 나누어 실시하였다. 그 결과 식이섭취량, 장기무게, 체중증가 정도에서 각 군에 따른 차이가 없는 것으로 나타났다. 7일간 투여 후 혈액 중 철분의 함량을 측정한 결과 결핍쥐에 Asp-Fe 투여군(175.2 ${\mu}g$/dL)과 heme-Fe 투여군(140.8 ${\mu}g$/dL)은 결핍 대조군(96.1 ${\mu}g$/dL)보다 유의적인 수준으로 증가하였다. 총 철분 결합능(total iron binding capacity, TIBC)를 측정한 결과 Asp-Fe 투여군(735.4 ${\mu}g$/dL)은 결핍 대조군(841.9 ${\mu}g$/dL)보다 유의적 수준으로 정상화되었다. 헤마토크리트(HCT) 수치를 측정한 결과에서 Asp-Fe 및 heme-Fe 모두 결핍 대조군보다 증가하는 경향은 보였지만 유의적인 차이는 없었다. 흡수율에서는 heme-Fe의 경우 21.3%인 반면에 Asp-Fe의 경우 50.2%로 약 2.3배 높은 것으로 나타났으며, 혈청에서의 철분농도 및 transferrin saturation(TS)는 heme-Fe 투여군 및 결핍 대조군에 비하여 Asp-Fe 투여군이 유의하게 높은 수준을 유지하였다. 이상의 결과로 미루어 볼 때 아미노산 킬레이트 철분은 heme-Fe과 유사한 수준의 생체 이용율을 가지고 있으며, 철분 결핍을 회복시키는데 매우 효과적인 보충제로서 이용될 수 있을 것으로 사료된다.

• Archives of Pharmacal Research
• /
• 제28권8호
• /
• pp.963-969
• /
• 2005

Many reports have stated that some of the pathogenic bacteria can obtain iron from ferroproteins, such as cytochrome C, ferritin, hemin, hemoglobin, and myoglobin. These reports prompted us to determine if an opportunistic pathogenic fungus, Candida albicans, can utilize ferroproteins to circumvent the iron-regulatory effect of transferrin. The following assays were carried out to measure in vitro growth stimulation by the ferroproteins: as an initial step, C. albicans was cultured in iron-free (pretreated with apotransferrin for 24h) culture medium. Once Candida albicans yeast cell growth reached stasis from iron starvation, individual ferroproteins were added to the culture media. Results showed that hemin, hemoglobin, and myoglobin supported a partial growth recovery. Additional studies with haptoglobin, a serum protein that interacts with the globin moiety of certain ferroproteins, established that C. albicans could obtain iron from the haptoglobin-ferroprotein complexes. These data indicate that the heme part of the ferroproteins is the source of iron. This implies that heme oxygenase, CaHMX1 might be involved in bringing about dissociation of heme-containing protein for iron-acquisition. In addition, anticandidal activity of transferrin takes place not only by the process of iron regulation, but also by direct interaction with the yeast cells.

• BMB Reports
• /
• 제45권4호
• /
• pp.239-241
• /
• 2012

Iron availability is limited in the environment and most bacteria have developed a system to acquire iron from host hemoproteins. Heme oxygenase plays an important role by degrading heme group and releasing the essential nutrient iron. The structure of Bacillus subtilis HmoB was determined to 2.0 ${\AA}$ resolution. B. subtilis HmoB contains a typical antibiotic biosynthesis monooxygenase (ABM) domain that spans from 71 to 146 residues and belongs to the IsdG family heme oxygenases. Comparison of HmoB and IsdG family proteins showed that the C-terminal region of HmoB has similar sequence and structure to IsdG family proteins and contains conserved critical residues for heme degradation. However, HmoB is distinct from other IsdG family proteins in that HmoB is about 60 amino acids longer in the N-terminus and does not form a dimer whereas previously studied IsdG family heme oxygenases form functional homodimers. Interestingly, the structure of monomeric HmoB resembles the dimeric structure of IsdG family proteins. Hence, B. subtilis HmoB is a heme oxygenase with a novel structural feature.

• 대한지역사회영양학회지
• /
• 제3권1호
• /
• pp.53-61
• /
• 1998

This study was designed to estimate mean daily iron intake and its bioavailabi- lity and to assess nutrition knowledge for 144 pregnant women in the last trimester. Serum ferritin concentration was analyzed to estimate their iron stores. Dietary intakes of iron(heme iron and nonheme iron), the amounts of MPF(meat, poultry and fish) and ascorbic acid were assessed by modified 24-hr recall method. The food frequency questionnaire was used to assess subjects usual food consumption patterns. The mean value of serum ferritin was $21.3\pm{15.2ng/ml}$ and 26.4% of the pregnant women had a serum ferritin level <12ng/ml(i.e. depleted iron stores). The mean daily intake of total orin in the pregnant women was 56.5%(17.0mg) of RDA and heme iron intake was 0.94mg which was 5.5% of total iron intake. Total absorbable iron calculated by the method of Monsen was 2.41mg and bioavailability of dietary iron was 2.41%. Food frequency test score of meats group was positively correlated(r=0.443) with the bioaavailability of dietary iron. The mean score on the nutrition knowledge test of subjects was 12.76(out of a possible 20 points). These results indicate that the nutritional iron status may be improved by increasing either the amount of iron in the diet or its availability.

• Archives of Pharmacal Research
• /
• 제29권3호
• /
• pp.249-255
• /
• 2006

Ability of iron acquisition of pathogenic microorganisms functions as a virulence factor. Candida albicans, a fungal pathogen that requires iron for growth, is susceptible to growth retardation by high-affinity iron binding proteins such as transferrin. Recently, we reported that C. albicans could utilize the heme as a part of heme-containing proteins dissociated by heme oxygenase, CaHMX1. In search of another pathway that C. albicans can use to bypass the growth regulation produced by iron limitation, this present study examined utilization of non-candidal siderophores such as Desferal and rhodotorulic acid (RA) for acquisition of inorganic iron by the fungus. C. albicans secreting no siderophores was cultured in iron-free (pretreated with apotransferrin for 24 h) (culture medium). Once growth of the yeast reached stasis from iron starvation, a siderophore was added to the culture media. Results showed that cultures containing apotransferrin within a dialysis membrane recovered growth to the level of untreated controls, whereas C. albicans yeast cells in direct contact with soluble iron-free (apo) transferrin recovered growth only partially. When static growth from iron limitation was reached, the addition of siderophore-apotransferrin complex to culture medium also permitted the yeast to recover growth from apotransferrin growth regulation. All the data show that C. albicans can utilize the non-candidal siderophores for iron acquisition under transferrin regulation as can pathogenic bacteria.