• Title/Summary/Keyword: Ammonium sulphate

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Presence of Rhodanese in the Cytosolic Fraction of the Fruit Bat (Eidolon helvum) Liver

  • Agboola, Femi Kayode;Okonji, Raphael Emuebie
    • BMB Reports
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    • v.37 no.3
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    • pp.275-281
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    • 2004
  • Rhodanese was isolated and purified from the cytosolic fraction of liver tissue homogenate of the fruit bat, Eidolon helvum, by using ammonium sulphate precipitation and CM-Sephadex C-50 ion exchange chromatography. The specific activity was increased 130-fold with a 53% recovery. The $K_m$ values for KCN and $Na_2S_2O_3$ as substrates were $13.5{\pm}2.2\;mM$ and $19.5{\pm}0.7\;mM$, respectively. The apparent molecular weight was estimated by gel filtration on a Sephadex G-100 column to be 36,000 Da. The optimal activity was found at a high pH (pH 9.0) and the temperature optimum was $35^{\circ}C$. An Arrhenius plot of the heat stability data consisted of two linear segments with a break occurring at $35^{\circ}C$. The apparent activation energy values from these slopes were 11.5 kcal/mol and 76.6 kcal/mol. Inhibition studies on the enzyme with a number of cations showed that $Mg^{2+}$, $Mn^{2+}$, $Ca^{2+}$, and $Co^{2+}$ did not affect the activity of the enzyme, but $Hg^{2+}$ and $Ba^{2+}$ inhibited the enzyme.

Characteristics of a Bifidobacterium longum LL04 ${\beta}$-Galactosidase (recombinant) Produced in Escherichia coli

  • Lim, Seong-Il;Kim, Geun-Bae;Yi, Sung-Hun;Lee, Byong-Hoon
    • Food Science and Biotechnology
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    • v.15 no.6
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    • pp.908-913
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    • 2006
  • Recombinant ${\beta}$-galactosidase from Bifidobacterium longum LL04 was expressed in Escherichia coli and partially purified by ammonium sulphate precipitation and anion-exchange chromatography (Mono-Q). The optimum temperature and pH of the partially purified enzyme were $50^{\circ}C$ and pH 7.0-8.0, respectively, when o-nitrophenyl-${\beta}$-D-galactopyranoside was used as a substrate. The enzyme was stable over the pH range of 5.0-9.0, and was active at $40^{\circ}C$ for more than 60 min at pH 7.0. The enzyme was significantly activated by $Na^+$ and $K^+$. Maximal activity was observed at the concentration of 10 mM for both $Na^+$ and $K^+$. The enzyme activity was strongly inhibited by most bivalent metal ions. The Km and Vmax on ONPG at 37 and $50^{\circ}C$ were 0.72, 167.9, and 0.507 mM, 310.9 U/mL, respectively.

Effect of Nitrate-Ammonium Ratio on Ionic Balance in Mulberry(Morus alba L.) Leaves (질산태(窒酸態) 및 암모니아태(態) 질소비율(窒素比率)과 상엽중(桑葉中)이온의 균형(均衡))

  • Lee, Won-Chu;Yoo, Sun-Ho;Lim, Sun-Uk
    • Korean Journal of Soil Science and Fertilizer
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    • v.15 no.2
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    • pp.110-116
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    • 1982
  • A water culture was conducted with 5 levels of nitrate-ammonium ratio (NAR: 10:0, 7:3, 5:5, 3:7, 0:10) to study the ionic balance, cation-anion (C-A) in mulberry leaves and to determine the optimum NAR for the greatest leaf yields. The results were as follows: 1. The growth rate of mulberry plants was the greatest at 7:3 NAR. 2. Magesium in the mulberry leaves was not affected by NAR. By lowering NAR, calcium content decreased more than potassium content increased, consequently lowering the sum of cation content (${\sum}C$). However, the sum of anion (${\sum}A$) increased because of increments of sulphate, chloride and especially phosphate. 3. Ionic balance in the mulberry leaves deceased from 727 to 116 me/Kg dry matter with increasing ammonium levels of 0 to 100% in the nutrient solution.

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$Fasciola$ $gigantica$ Fatty Acid Binding Protein (FABP) as a Prophylactic Agent against $Schistosoma$ $mansoni$ Infection in CD1 Mice

  • Aly, Ibrahim Rabia;Diab, M.;El-Amir, A.M.;Hendawy, M.;Kadry, S.
    • Parasites, Hosts and Diseases
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    • v.50 no.1
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    • pp.37-43
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    • 2012
  • Although schistosomicidal drugs and other control measures exist, the advent of an efficacious vaccine remains the most potentially powerful means for controlling this disease. In this study, native fatty acid binding protein (FABP) from $Fasciola$ $gigantica$ was purified from the adult worm's crude extract by saturation with ammonium sulphate followed by separation on DEAE-Sephadex A-50 anion exchange chromatography and gel filtration using Sephacryl HR-100, respectively. CD1 mice were immunized with the purified, native $F.$ $gigantica$ FABP in Freund's adjuvant and challenged subcutaneously with 120 $Schistosoma$ $mansoni$ cercariae. Immunization of CD1 mice with $F.$ $gigantica$ FABP has induced heterologous protection against $S.$ $mansoni$, evidenced by the significant reduction in mean worm burden (72.3%), liver and intestinal egg counts (81.3% and 80.8%, respectively), and hepatic granuloma counts (42%). Also, it elicited mixed $IgG_1/IgG_{2b}$ immune responses with predominant $IgG_1$ isotype, suggesting that native $F.$ $gigantica$ FABP is mediated by a mixed Th1/Th2 response. However, it failed to induce any significant differences in the oogram pattern or in the mean granuloma diameter. This indicated that native $F.$ $gigantica$ FABP could be a promising vaccine candidate against $S.$ $mansoni$ infection.

Studies on the Microbial Utilization of Agricultural Wastes (Part 11) Properties of Cellulolytic Enzyme Produced by a Cellulolytic Fungus Trichodrma sp. KI 7-2 and its Application to the Fermented Feed Production (농산폐자원의 미생물학적 이용에 관한 연구(제11보) Trichoderma sp KI 7-2가 생산하는 섬유소분해효소의 성질 및 발효사요에의 응용)

  • Bae, Moo;Lee, Gye-Jun;Tak, Sun-Mi;Kim, Byung-Hong
    • Microbiology and Biotechnology Letters
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    • v.6 no.1
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    • pp.1-8
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    • 1978
  • In order to develop the processes for the production of fermented feed from cellulosic agricultural by-product, cereal straw, by th action of cellulolytic fungus, the properties of the cellulolytic enzyme produced by Trichoderma sp. KI 7-2 was studied. A higher enzyme activity was obtained in the culture added by 1% rice or barley straw powder than in the culture of pure cellulose. The crude enzyme was prepared by precipitating from 20∼60% saturated ammonium sulphate of the culture supernatant. The optimum conditions for the enzyme reaction were temperature of of 50$^{\circ}C$ and pH 4.2. The crude enzyme was static at 50$^{\circ}C$ for two hours and at pH between 4 and 6. These properties were adopted for the fermented feed production, and several production. Thus, several processes of semisolid culture were devicced to up grade tile fermented feed and to develop into the acceptable quality.

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Effect of hCG on TeBG (hCG가 TeBG에 미치는 영향)

  • Sung, Ho-Kyung;Kim, Woo-Gyeum
    • The Korean Journal of Physiology
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    • v.14 no.1
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    • pp.7-13
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    • 1980
  • In the previous experiment, authors have shown that during the latter half of estrous cycle there was an increase in plasma testosterone level in the rats stimulated with hCG. To determine the physiologic significance of elevated plasma testosterone, changes of the plasma concentrations of TeBG and testosterone following hCG stimulation were analyzed in the rats having a regular 5 day cycle. The rats were divided into three groups; the control, the rats stimulated with single hCG on the day of proestrus and stimulated with hCG throughout the entire cycle. Blood samples were obtained once a day for an estrous cycle and analyzed for the binding capacity of TeBG using ammonium sulphate precipitation method and testosterone concentration by means of radioimmunoassay. Followings were the results; 1) There was no significant variation in the binding capacity of TeBG in peripheral blood during the estrous cycle of the control rats. 2) No cyclic variation in the binding capacity of TeBG was observed in the rats stimulated with single hCG on proestrus. although the levels tended to be higher in the rats with stimulation than in the control rats. 3) Continual stimulation of hCG produced a marked increase in the binding capacity of TeBG especially on the day of metaestrus. 4) The changes in the plasma level of testosterone followed the same basic pattern seen in the TeBG binding capacity. 5) From above results, the followings were suggested. a. hCG related increase of the binding capacity of TeBG is probably secondary to a modest increase in estrogen as well. b. hCG related increase of plasma testosterone in female rats is not entirely due to excess production rather in part due to decreased metabolism induced by the rise in TeBG. c. It seems likely that most of elevated testosterone shown in the rat stimulated with hCG is bound to TeBG and only small portion is unbound form which influence cellular activity. It is rather possible that an increase in TeBG could augment estrogen activity.

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Evaluation of protective coatings for geopolymer mortar under aggressive environment

  • Rathinam, Kumutha;Kanagarajan, Vijai;Banu, Sara
    • Advances in materials Research
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    • v.9 no.3
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    • pp.219-231
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    • 2020
  • The aim of this study is to investigate the durability of fly ash based geopolymer mortar with and without protective coatings in aggressive chemical environments. The source materials for geopolymer are Fly ash and Ground Granulated Blast furnace Slag (GGBS) and they are considered in the combination of 80% & 20% respectively. Two Molarities of NaOH solution were considered such as 8M and 10M. The ratio of binder to sand and Sodium silicate to Sodium hydroxide solution (Na2SiO3/NaOH) are taken as 1:2 and 2 respectively. The alkaline liquid to binder ratio is 0.4. Compressive strength tests were conducted at various ages of the mortar specimens. In order to evaluate the performance of coatings on geopolymer mortar under aggressive chemical environment, the mortar specimens were coated with two different types of coatings such as epoxy and Acrylic. They were then subjected to different chemical environments by immersing them in 10% standard solutions of each ammonium nitrate, sodium chloride and sulphuric acid. Drop in compressive strength as a result of chemical exposure was considered as a measure of chemical attack and the drop in compressive strength was measured after 30 and 60 days of chemical exposure. The compressive strength results following chemical exposure indicated that the specimens containing the acrylic coating proved to be more resistant to chemical attacks. The control specimen without coating showed a much greater degree of deterioration. Therefore, the application of acrylic coating was invariably much more effective in improving the compressive strength as well as the resistance of mortar against chemical attacks. The results also indicated that among all the aggressive attacks, the sulphate environment has the most adverse effect in terms of lowering the strength.

Effect of Follicular Fluid Proteins and Gonadotropins on Progesterone Secretion by Buffalo Granulosa Cells In vitro

  • Vinze, Mukesh;Sharma, M.K.;Singh, Dheer
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.11
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    • pp.1496-1500
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    • 2004
  • In the mammalian ovary the follicular fluid contains proteins and peptides which play an important role in growth, development and maturation of oocytes. The gonadotropins and some other factors work synergistically and regulate ovarian functions. In the present study the effect of follicular fluid proteins (FFP) and gonadotropins on progesterone secretion by granulosa cells (GC) from buffalo ovary, was investigated during culture. The follicular fluid was collected from small (<5 mm), and medium (5-8 mm) follicles obtained from buffalo ovaries. The follicular fluid from medium follicles was fractionated with ammonium sulphate at 80% saturation. The precipitated protein fraction was further resolved in to minor (peaks I, III) and major (peak II) proteins using gel filtration (Sephadex G-200). The FFP from small follicles and major FFP (peak II) at a dose of 200 $\mu$g/well, significantly stimulated progesterone secretion by pooled GC (3${\times}10^{5}$ cells/2 ml medium/well). The minor FFP did not show any stimulatory effect. There was a significant increase in progesterone secretion by pooled GC in presence of FFP and LH (10 ng/well), however, FSH (20 ng/well) with FFP exhibited an inhibitory effect. The major FFP and gonadotropins were also studied for their effect on progesterone production by GC isolated from medium and large size follicles. The GC from medium follicles were more responsive to FSH and FFP whereas GC from large follicles exhibited enhanced progesterone secretion with LH and FFP. These results indicated that FFP have their own stimulatory effect and also act synergistically with gonadotropins. The significantly different response shown by GC, for steroid hormone secretion, is based on their stage of growth and differentiation. The purification and characterization of such steroidogenic proteins may help in elucidating their role in growth and differentiation of granulosa cells.

Ovarian Response and Profile of Plasma Sex Steroids in Goats Against Combined Administration of FSH and LH Isolated from the Pituitaries of Buffaloes

  • Taru Sharma, G.;Pande, J.K.;Sanwal, P.C.;Varshney, V.P.
    • Asian-Australasian Journal of Animal Sciences
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    • v.10 no.5
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    • pp.514-518
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    • 1997
  • This study was designed to record the ovarian response towards a combined administration of heterologous buffalo FSH (buFSH) and LH (buLH) in goats. The impact of such a treatment on ovarian structures and on the plasma profile of the ovarian sex steroids (estradiol $17-{\beta}$ and progesterone) was studied. The buFSH and buLH were isolated from the buffalo pituitaries involving a procedure of ethanolic extraction, acetone precipitation followed by metaphosphoric acid - ammonium sulphate fractionation. Both gonadotrophin samples prepared were found biologically active and potent. There was an increase in the total number of follicles in the treated group ($12.66{\pm}1.24$) vis-a-vis the control group ($8.50{\pm}2.06$). However, the percentage ($51.48{\pm}6.37$) of large follicles were found reduced ($23.74{\pm}5.93$) following the treatment. Again the number of corpora lutea were observed significantly higher ($2.33{\pm}0.47C.L.$) in the treated group than (1 C. L.) in the control group. The peak plasma estradiol- $17{\beta}$ levels achieved, were much higher ($17.16{\pm}9.52pg/ml$) in the treated group, than the peak ($7.22{\pm}1.67pg/ml$) achieved in the control group. Similar trend was observed with respect to the progesterone levels (higher in the treated group). This study thus indicated that, a combined administration of heterologous buffalo FSH and LH to goats speeded up development of larger follicles nearing the ovulation stage. This population of the follicles subsequently got reduced and lead to the formation of the increased number of the corpora lutea observed in this study.

Characteristics of Urease from Vibrio parahaemolyticus Possessing tah and the Genes Isolated in Korea

  • Kim, Young-Hee;Kim, Jong-Sook
    • Journal of Microbiology
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    • v.39 no.4
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    • pp.279-285
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    • 2001
  • Vibrio parahaemolyticus is a halophilic bacterium associated with seafood gastroenteritis. An unusual strain of Kanagawa-positive urease producing Vibrio parahaemolyticus O1:K1 was isolated from the environment and identified . A polymerase chain reaction assay revealed that this strain harbored both the tdh and the genes. The urease from this strain was studied. Maximum urease production was induced in LB medium containing 0.2% urea, 0.5% glucose, 2% NaCl and pH 5.5 with 6h of culti-vation at 37$\^{C}$ under aeration. Purification of urease was achieved by the process of whole cell lysate, 65% ammonium sulphate precipitation, DEAE-cellulose ion exchange column chromatography, Sepharose CL-6B gel filtration and oxirane activated Sepharose 6B-urea affinity chromatography with 203 fold purification and 2.2% yield. Analysis of the purified enzyme by SDS-PAGE demonstrated the presence of the subunits with a molecular weight of 85kDa, 59kDa, 41kDa and the molecular weight for the native enzyme by nondenaturing PAGE and gel filtration chromatography was 255kDa. The purified urease was stable at pH 7.5 and the opeimal pH in HEPES buffer was 8.0 The enzyme was stable at 60$\^{C}$ for 2 h with a residual activity of 32% . The addition of 10$\mu$M if NiCl$_2$maintained stability for 30 min. The Km value of the purified enzyme was 35.6 mM in urea substrate. The TD$\_$50/(median toxic dose) of the purified urease was 2.5$\mu\textrm{g}$/ml on human leukemia cells.

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