• Applied Biological Chemistry
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• v.44 no.4
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• pp.219-223
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• 2001

Effects of hemoglobin (Hgb) concentration and degree of hydrolysis (DH) of Hgb on the separation of heme-iron were examined to produce highly enriched heme-iron from Hgb hydrolysate. Separation efficiency of Hgb hydrolysate with different DH was studied at wide pH range (pH $1.0{\sim}11.0$). Separation efficiency expressed as heme-iron/peptide ratio increased with decreasing Hgb concentration. When 5% Hgb (pH 10.0) was hydrolyzed using commercially available Esperase for 5 h at $50^{\circ}C$, DH was 25%. The precipitation of heme-iron-enriched peptides were remarkably high at pH range $3{\sim}6$. Optimal pH range for heme-iron with high heme-iron/peptide ratio shifted to acidic pH with increasing DHs of Hgb. The enriched heme-iron fraction in the precipitates showed a single band through urea-SDS-PAGE, with a molecular mass of 1 kDa. In the dry heme-iron product produced in a pilot bioreactor, content of heme-iron and heme-iron/peptide ratio were 27.1 and 38.7%, respectively, and production yield was 9.3%.

• Applied Biological Chemistry
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• v.46 no.2
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• pp.130-133
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• 2003

A method for separating heme-iron from hemoglobin (Hgb) hydrolysate by dialysis was developed. Recovery of heme-iron increased with increasing Hgb concentration, whereas rejection of peptide and separation effciency expressed by HP ratio (heme-iron/peptide) did not show significant differences. HP ratio increased with increases in the degree of hydrolysis of Hgb and $KH_2PO_4$ concentrations of dialysis solution. Recovery of heme-iron decreased with increase in the pH of dialysis solution due to wash-out of heme-iron across the dialysis membrane caused by increase in solubility of heme-iron. Rejections of peptide were 74.5 and 87.5% (2 and 5 kDa of cut off size, respectively), whereas recovery of heme-iron decreased from 86.5 (2 kDa) to 63.1% (25 kDa). Amounts of heme-iron and peptide of dried heme-iron product were 21.7 and 77.0%, and HP ratio and production yield were 28.2 and 6.5%, respectively.

• Journal of Microbiology and Biotechnology
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• v.19 no.6
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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.

• Nutritional Sciences
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• v.9 no.4
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• pp.295-300
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• 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.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.35 no.2
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• pp.162-165
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• 2002

To determine changes of heme compounds on lipid oxidation during repeat heating in white muscled fish (yellowfin sole and yellow croaker), myoglobin, metmyoglobin, total iron, nonheme iron and heme iron contents were analysed. Myoglobin content was decreased in the step of repeat heating. Especially, it was decreased the most rapidly roasted at 180$^{\circ}C$ for 20 min in fillet samples. The skinless fillet roasted at the lower temperature resulted in the higher level of metmyoglobin associated with the reduced myoglobin. Regardless of roasted temperature and time, total iron content was not change in contrast of raw meat throughout processing. Nonheme iron content was increased, but heme iron content was decreased during roasted, heated and reheated.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.12
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• pp.1720-1725
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• 2011

Aspartic acid chelated iron (Asp-Fe) was synthesized by a new method using calcium carbonate, aspartic acid, and ferrous sulfate. This study was carried out to investigate the bioavailability of Asp-Fe in iron-deficient rats. We divided the rats into four experimental groups. The first was the normal diet control group, or NC. The second was the no treated control group of iron-deficient (ID) rats, or ID+C. The third was the heme-iron (heme-Fe) treated group of ID rats, ID+heme-Fe. And the fourth was the Asp-Fe treated group of ID rats, or ID+Asp-Fe. There were no differences among any of the experimental groups in diet consumption, change of body weight, or the weight of the livers, kidneys, or spleens. After 7 days of feeding, the iron content in the sera of the ID+Asp-Fe group (175.2 ${\mu}g$/dL) and the ID+heme-Fe group (140.8 ${\mu}g$/dL) were significantly higher than that of the ID-C group (96.1 ${\mu}g$/dL). The total iron binding capacity (TIBC) of the ID+Asp-Fe group (735.4 ${\mu}g$/dL) was significantly normalized compared to the ID+C group (841.9 ${\mu}g$/dL) or ID+heme-Fe group (824.6 ${\mu}g$/dL). The hematocrit level of the ID+Asp-Fe group was increased to normal levels, but there was no statistical difference among ID groups. The absorption ratio of heme-Fe was 21.3% and that of Asp-Fe was 50.2%, which indicates a 2.3 times higher ratio in comparison with heme iron. With the above results we found that Asp-Fe seems to be an efficient form of iron to supply iron deficient rats in order to cure them of anemia. Thus, these findings suggest that aspartic acid chelated iron has the potential to serve as a functional food related to iron metabolism.

• Archives of Pharmacal Research
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• v.28 no.8
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• pp.963-969
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• 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
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• v.45 no.4
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• pp.239-241
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• 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.

• Korean Journal of Community Nutrition
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• v.3 no.1
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• pp.53-61
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• 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
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• v.29 no.3
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• pp.249-255
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• 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.