• Title/Summary/Keyword: Binding Capacity

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Influence of Growth Rate on Biosorption of Heavy Metals by Nocardia amarae

  • Kim, Dong Wook;Daniel K. Cha;Hyung-Joon Seo;Jong Bok Bak
    • Journal of Microbiology and Biotechnology
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    • v.12 no.6
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    • pp.878-881
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    • 2002
  • The goal of the current research was to assess the influence of the growth rate of Nocardia amarae on its overall metal binding capacity. Batch sorption isotherms for cadmium (Cd), copper (Cu), and nickel (Ni) showed that Nocardia cells harvested from chemostat cultures at a dilution rate of $0.33d^-1$ had a significantly higher metal sorption capacity than cells grown at 0.5 and $1d^-1$. The cell surface area estimated using a dye technique indicated that pure N. amarae cells grown at a lower growth rate had a significantly more specific surface area than cells harvested from a higher growth rate operation. Accordingly, this difference in the specific surface area seemed to indicate that the higher metal sorption capacity of the slowly growing Nocardia cells was due to their higher specific surface area.

Effects of Commercial Soy Milk and Cow Milk on Iron Status and Work Capacity of Rats (시판 두유 및 우유가 흰쥐의 체내 철분 영양상태와 운동능력에 미치는 영향)

  • 이윤복
    • Journal of Nutrition and Health
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    • v.30 no.8
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    • pp.904-910
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    • 1997
  • Evaluation of soy milk as an iron-rich milk substitute was conducted by feeding commercial soy milk , cow milk and mixed formula(soy and cow milk) to Wistar rats for 8 weeks. Body weight gains were significantly lower in the soy milk and mixed formula groups. Hematocrit, serum iron concentration and TIBC (total iron binding capacity) were measured to determine the iron status of the rats. In these respects, the iron status of the soy milk group was normal. Both serum iron concentration and TIBC as well as hematocrit were abnormal in the cow milk group , which is indictive of severe iron deficiency . Although hematocrit was normal in the mixed formula group, serum iron concentration was lightly low. The work capacities of each group were correlated with serum iron concentration and tIBC rather than hematocrit. The running distance of the soy milk group was about 10 -fold longer than that of the cow milk group. Soy milk may be considered an iron- rich substitute for cow milk due to its higher iron content and bioavailability.

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Effect of Chlorella on Metallothionein Synthesis and Binding Capacity of Cadmium in Cadmium Poisoned Rat Liver and Kidney

  • Hwang Yoo-Kyeong;Choi Hyun-Jin;Nan Meng;Yoo Jai-Du;Kim Yong-Ho
    • Biomedical Science Letters
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    • v.12 no.1
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    • pp.23-27
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    • 2006
  • The rate of metallothionein synthesis on cadmium-poisoned rats reflects the level of toxicity, and also it reduces the toxicity which is caused by the uptake of cadmium. Chlorella supplementation in the diets of the cadmium-poisoned rats decreased the concentration of cadmium in blood and urine compared with the control group. Although the liver and kidneys of rats are major target organs of cadmium and coherence of metallothionein and cadmium, no previous study has determined the correlation between the rate of metallothionein synthesis in the liver and kidneys of rats and dietary supplementation of chlorella with cadmium uptake. This study analyzed total metallothionein level on the tissue of the liver and kidneys, the concentration of cadmium bound to the metallothionein, and the total concentration of cadmium on the tissue of the liver and kidneys after dietary supplementation with 1%, 5%, and 10% dried chlorella and 40 ppm of cadmium to 46 male SD rats (mean weight: $150\pm20\;g$) for 4 weeks. According to the data analysis of the total rate of metallothionein synthesis in the liver and kidneys, the group of SD rats on the supplementation with 1% chlorella and 40 ppm of cadmium showed a rate of $93.2\pm8.9\;ng/g$, a significant decrease of 58.8% compared to that of the control group of SD rats on the supplementation with cadmium only, which showed a rate of $227.3\pm32.5 ng/g$ (P=0.0001). In contrast, no significant difference was observed through the changing of chlorella concentrations between 5% and 10% chlorella supplementation with cadmium. The group supplemented with 1% or greater chlorella levels represented a greater decrease in the total cadmium concentration of the kidney and liver tissues, the amount of total metallothionein synthesis, the amount of metallothionein with binding to cadmium, and the concentration of free cadmium without binding to metallothionein. Consequently, the supplementation of 1% and 5% chlorella was effective in reducing the synthesis of metallothionein for cadmium uptake, but increased the rate of binding of cadmium to metallothionein.

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Differential Sialic Acid Content and Hemoglobin-binding Activity of Precursor Prohaptoglobin and Mature Haptoglobin (전구체 프로합토글로빈과 성숙 합토글로빈의 시알산 함량 및 헤모글로빈-결합력 비교)

  • Lee, Joo-Hyun;Oh, Mi-Kyung;Kim, In-Sook
    • Journal of Life Science
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    • v.27 no.6
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    • pp.632-639
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    • 2017
  • Mature haptoglobin (Hp) is a plasma glycoprotein and acts as an antioxidant by scavenging cell-free hemoglobin (Hb). Prohaptoglobin (proHp) is an unprocessed Hp precursor which is present a little in circulation. However, the biological function of proHp remains unknown. To investigate the structural and functional differences between proHp and Hp, we prepared recombinant proHp isoforms and compared their sialic acid content and Hb-binding capacity with those of mature isoforms. When proHp samples were analyzed by Western blot under non-reducing conditions, proHp1 was detected as one band of approximately 130 kDa and proHp2 as multiple bands >200 kDa, in the manner of mature Hp1-1 and Hp2-2, respectively. On the native polyacrylamide gel under non-reducing and non-denaturing conditions, both proHp isoforms migrated more slowly than their mature Hp counterparts. In addition, the lectin-based ELISA assay demonstrated that the content of sialic acid in proHp1 and proHp2 was much less than in Hp1-1 and Hp2-2. The Hb-binding capacity of proHp was also lower than those of mature Hp. These findings indicate that proHp and Hp are similar in the size and polymerization pattern, but different in sialic acid content and Hb-binding activity. It suggests precursor proHp may exert different functions in circulation than does mature Hp.

Fusobacterium nucleatum modulates serum binding to Porphyromonas gingivalis biofilm (Porphyromonas gingivalis biofilm에 대한 면역혈청의 침투력에 대한 Fusobacterium nucleatum의 조절효과)

  • Choi, Jeom-Il;Kim, Sung-Jo;Kim, Soo-Jin
    • Journal of Periodontal and Implant Science
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    • v.31 no.4
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    • pp.661-668
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    • 2001
  • Anti-P. gingivalis immune sera were obtained from mice immunized with either P. gingivalis alone, or F. nucleaturm followed by P. gingivalis. Two groups of immune sera were examined for binding capacity to P. gingivalis biofilm by confocal laser scanning microscope, Antibody avidity index was also determined for each immune sera. The results indicated that prior immunization of mice with F. nucleaturm impaired P. gingivalis-specific immune sera in binding capacity to biofilm and antibody avidity to P. gingivalis. Elevated antibody responses in patients with destructive periodontal disease has often been related to suboptimal level of protective antibody $(opsonophagocytosis)^{1-3)}$ while post-immune sera obtained with experimental animals using a single periodontal pathogen demonstrated satisfactory levels of protective function against the homologous bacterial $challenge^{4,5)}$.The reason is unclear why elevated IgG responses in periodontal patients to periodontal pathogens do not necessarily reflect their protective function. Such an immune deviation might be derived from the fact that destructive periodontal disease is cumulative result of immunopathologic processes responding to an array of different colonizing microorganisms sequentially infecting in the subgingival environmental niche. Fusobacterium nucleaturm is one of the key pathogens in gingivitis, in the transitional phase of conversion of gingivitis into destructive periodontitk, and in adult $periodontitis^{6-8)}$. It also plays a central role in coaggregation with other important microbial species in subgingival $area^{6,9,10)}$ as well as in $biofilm^{11)}$, especially with Porphyromonas gingjvalis in synergism of virulence in human periodontal disease or in animal $models^{12-14)}$. This organism has also been reported to have immune modulating activity for secondary immune response to Actinobacillus $actinomycetemcomitans^{15)}$. It is presumed that sequential colonization and intermicrobial coaggregation between intermediate and late colonizers could potentially modulate the immune responses and development of specific T cell phenotypes in periodontal lesions. We have recently demonstrated the skewed polarization of P. gingivalis-specific helper T cell clones in mice immunized with F. nucleaturm followed by P. $gingivalis.^{16)}$. Consequently F. nucleaturm may initially prime the immune cells and modify their responses to the successive organism, P. gingivalis. This could explain why one frequently observes non-protective serum antibodies to P. gingivalis in periodontal patients in contrast with those obtained from animals that were immunized with $P.gingivalis\;alone^{17)}$. The present study was performed to investigate the immune modulating effect of F. nucleatum on serum binding to experimental biofilms and the avidity of anti-P. gingivalis antibody.

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Increased Expression of the ${\alpha}_2$ Isoform of (Na,K)ATPase in the Differentiated Murine Muscle Cell Line BC3H-1 (BC3H-1 분화세포에서의 (Na,K)ATPase ${\alpha}_2$ isoform의 표현증대)

  • Lee, Kyung-Lim
    • YAKHAK HOEJI
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    • v.40 no.6
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    • pp.734-738
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    • 1996
  • The development of the alpha2 isoform of (Na,K)ATPase which is high affinity ouabain receptors was studied in the differentiating nonfusing muscle cell line BC3H-1. T he differentiation process of BC3H-1 cell line was confirmed by 2-dexy-D-[$^3$H] glucose uptake experiment and the quantity of the expression of ${\alpha}_2$ isoform was measured using a whole cell [$^3$H] ouabain-binding assay. Undifferentiated growing BC3H-1 cells, myoblasts, exhibited low levels of insulin-stimulated glucose uptake and [$^3$H] ouabain-binding sites. In contrast, differentiated BC3H-1 cells, myocytes, had a 5.6-fold increase in insulin-stimulated glucose uptake and 5-fold increase in [$^3$H] ouabain-binding sites. Scatchard analysis showed that myocytes developed more [$^3$H] ouabain-binding sites than myoblasts vath a dissociation constant (kd) of 6${\times}10^{-8}$M and capacity of 6.l${\times}10^{-5}$ sites/cell. Therefore. it seems that myoblasts express low levels of ${\alpha}_2$ subunit and probably the majority of ${\alpha}_1$ subunit, whereas myocytes express high levels of ${\alpha}_2$ isoform. The results indicate that the expression of ${\alpha}_2$ isoform is developmentally regulated during differentiation and that BC3H-1 culture system provides an excellent model for the study of differentiation and mechanism of (Na,K)ATPase action in muscle which requires electrical excitability.

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Manufacturing of Calcium Binding Peptide using Sericin Hydrolysate and Its Bioavailability in Calcium Deficient Rat (실크 세리신 단백질 가수분해물을 이용한 유기 칼슘제의 제조 및 칼슘 결핍 쥐에서의 생체 이용률)

  • Cho, Hye-Jin;Lee, Hyun-Sun;Jung, Eun-Young;Suh, Hyung-Joo
    • Journal of the East Asian Society of Dietary Life
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    • v.20 no.5
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    • pp.680-686
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    • 2010
  • Silk sericin protein was hydrolyzed by seven proteolytic enzymes in order to examine the effectiveness of the hydrolysates in binding calcium. The amino acid nitrogen content of hydrolysates from Flavourzyme was higher than that for other enzymes, and its calcium binding capacity showed a dose-dependent increase. We examined the effects of calcium binding peptide from sericin hydolysates on the bioavailability of Ca-deficient rats. Three-week-old male rats were fed an Ca-deficient diet for three weeks. Rats were divided into four groups (DD: non-treated group on calcium deficient diet; DD+MC: milk-calcium treated group; DD+OC: organic calcium made using sericin hydolysates; and DD+IC: inorganic calcium ($CaCl_2$). After oral administration of calcium supplements for one week, the calcium content of the serum and liver were significantly higher in DD+OC ($101.7{\mu}g$/mL and $49.3{\mu}g$/mL) and DD+MC ($83.6{\mu}g$/mL and $42.8{\mu}g$/mL) than DD ($86.3{\mu}g$/mL and $43.4{\mu}g$/mL). The alkaline phosphatase (ALP) content in the treated groups was significantly lower than DD, but no significant difference among groups was shown. Aspartate aminotransferase (AST) levels did not show any significant difference between groups. Alanine aminotransferase (ALT) levels were significantly reduced compared to the DD group. In conclusion, binding calcium to peptides from sericin hydrolysates seems to improve its bioavailability, and to hasten the cure of calcium deficiency in experimental rats.

Antibiofilm Activity and Binding Specificity of Polyclonal DNA Aptamers on Staphylococcus aureus and Escherichia coli

  • Arizah Kusumawati;Apon Zaenal Mustopa;Rifqiyah Nur Umami;Adi Santoso;I Wayan Teguh Wibawan;Agus Setiyono;Mirnawati Bachrum Sudarwanto
    • Microbiology and Biotechnology Letters
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    • v.50 no.3
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    • pp.328-336
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    • 2022
  • Aptamers are short, chemically synthesized, single-stranded DNA or RNA oligonucleotides that fold into unique three-dimensional structures. In this study, we aim to determine the antibiofilm activity and binding specificity of the six polyclonal DNA aptamers (S15K3, S15K4, S15K6, S15K13, S15K15, and S15K20) on Staphylococcus aureus BPA-12 and Escherichia coli EPEC 4. Aptamer S15K6 showed the highest percentage of antibiofilm activity against S. aureus BPA-12 (37.4%) as shown by the lowest OD570 value of 0.313. Aptamer S15K20 showed the highest percentage of antibiofilm activity against E. coli EPEC 4 (15.4%) as shown by the lowest OD570 value of 0.515. Aptamers S15K13 and S15K20 showed antibiofilm activities against both S. aureus BPA-12 and E. coli EPEC4, and thus potentially have broad reactivity. Furthermore, based on the binding capacity and Kd values from our previous study, the binding specificity assay of selected polyclonal DNA aptamers (S15K3 and S15K15) against S. aureus BPA-12, E. coli EPEC 4, S. aureus BPA-6, S. agalactiae, E. coli MHA-6, and Listeria monocytogenes were performed using qPCR. Aptamers S15K3 and S15K15 showed specific binding to S. aureus BPA-12, E. coli EPEC 4, S. aureus BPA-6, and S. agalactiae, but could not bind to E. coli MHA-6 and L. monocytogenes. Therefore, this study showed that the polyclonal DNA aptamers have antibiofilm activity and were able to bind to S. aureus BPA-12 and E. coli EPEC 4 bacteria.

Analysis of calmodulin binding property of IQ motifs of IQGAP1 (IQGAP1내에 존재하는 IQ 부위들의 CaM 결합 특성 분석)

  • Jang, Deok-Jin
    • Analytical Science and Technology
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    • v.24 no.6
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    • pp.527-532
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    • 2011
  • IQ motif-containing GTPase-activating protein 1 (IQGAP1), which is a well-known $Ca^{2+}$-independent calmodulin (CaM) binding protein, is involved in various cellular functions such as cell proliferation and cell migration. IQGAP1 has four repeated IQ motifs, which are crucial for CaM binding. It has been shown that all four IQ motifs of IQGAP1 can bind to $Ca^{2+}$/CaM, while the third and fourth IQ motifs of IQGAP1 can bind to apoCaM. However, it has not been clear whether the CaM binding of IQ motifs of IQGAP1 was mediated directly or indirectly. In this study, we examined whether the binding between CaM and each IQ motif of IQGAP1 was direct in vitro. As a result, we found that IQ1 motif has a weak $Ca^{2+}$-dependent CaM binding. In contrast, IQ3 has a $Ca^{2+}$-dependent CaM binding. All other motifs have no significant CaM binding. We also found that IQ(2.7-3) and IQ(3.5-4.4) have CaM binding capacity. This finding indicates that IQ motifs of IQGAP1 plays a dynamic role via different motif interactions with $Ca^{2+}$/CaM or proCaM.

Heterocyclic Amines Removal by Binding Ability of Lactic Acid Bacteria Isolated from Soybean Paste (된장에서 분리된 유산균의 결합력에 의한 Heterocyclic Amines 제거)

  • Lim, Sung-Mee
    • Korean Journal of Microbiology
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    • v.50 no.1
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    • pp.73-83
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    • 2014
  • The objective of the this study was to investigate the binding capacity and removal ability of lactic acid bacterial strains obtained from Korean soybean paste for mutagenic heterocyclic amines (HCAs) formed during cooking of protein-rich food at high temperature. Among 19 strains identified by carbohydrate fermentation and 16S rRNA sequencing, the live cell or cell-free culture supernatant of Lactobacillus acidophilus D11, Enterococcus faecium D12, Pediococcus acidilactici D19, L. acidophilus D38, Lactobacillus sakei D44, Enterococcus faecalis D66, and Lactobacillus plantarum D70 inhibited the mutagenesis caused by either 3-amino-1,4-dimethyl-5H-pyrido[4,3-b] indole (Trp-P-1) or 3-amino-1-methyl-5H-pyrido[4,3-b] indole (Trp-P-2) in Salmonella typhimurium TA98 and TA100. The bacterial cells of the isolated strains showed greater binding activity than the pure cell wall, exopolysaccharide, and pepetidoglycan. The carbohydrate moieties of the cell wall or protein molecules on the cell surface have a significant role in binding Trp-P-1 and Trp-P-2, since protease, heating, sodium metaperiodate, or acidic pH treatments significantly (P<0.05) reduced the binding efficacy of the tested bacteria. Addition of metal ions or sodium dodecyl sulfate decreased the binding ability of E. faecium D12, L. acidophilus D38, and E. faecalis D66. Therefore, the binding mechanisms of these strains may consist of ion-exchange and hydrophobic bonds. Especially, the high mutagen binding by L. acidophilus D38 and L. plantarum D70 may reduce the accumulation or absorption of Trp-P-1 and Trp-P-2 in the small intestine via increased excretion of a mutagen-bacteria complex.