• Asian-Australasian Journal of Animal Sciences
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• v.9 no.6
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• pp.701-703
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• 1996

The influence of phenylalanine (Phe) in the medium on protein synthesis of chicken embryo fibroblasts (CEF) was examined. CEF was derived from 9-d-old embryos by trypsin-EDTA digestion. To examine the deficiency of Phe in the medium, CEF was cultured in Dulbecco's modified Eagle's medium (DMEM) with or without Phe. CEF was also cultured in Dulbecco's phosphate buffered saline (PBS ($Ca^{2+}$, $Mg^{2+}$)) with or without $400{\mu}m$ Phe in order to examine the effect of Phe supplementation. All media were supplemented with 10% (v/v) fetal calf serum. After incubation for 6, 30 and 54 h, protein synthesis was measured by the incorporation of L-[2, $6-^{3}H$] Phe into CEF for further 18 h. Protein synthesis of CEF cultured in DMEM was higher than that in PBS ($Ca^{2+}$, $Mg^{2+}$). High specific radioactivity of Phe due to the low concentration of Phe in the medium resulted in the apparent increase in protein synthesis of CEF. Protein synthesis cultured in PBS ($Ca^{2+}$, $Mg^{2+}$) with Phe did not increase during 72 h of cell culture.

• Biomolecules & Therapeutics
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• v.9 no.1
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• pp.40-45
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• 2001

The activity of taurine transporter is affected by various extracellular stimuli such as ion, hormone and stress. To assess effects of steroid hormones antral cyclosporin A (CsA) on the taurine transporter activity, murine monocytic RAW264.7 cell line was stimulated with dexamethasone (DM), triamcinolone (TA), cortisone (CS), hydrocortisone (HCS), prednisone (PSN), prednisolone (PSL) and methylprednisolone (MPSL) in the presence of 12-0-tetradecanoylphorbol-13-acetate(TPA). Treatment of TPA on the cell line led to significant reduction of taurine transporter activity. However, in case of stimulation of the cells with steroid hormones in the presence of TPA, all of them recovered TPA-induced reduction of the taurine transporter activity. Treatment of the cells with CsA led to significant reduction of the taurine transporter activity. Ionomycin (IM) recovered the reduced taurine transporter activity by CsA, but failed in the presence of EDTA, a calcium chelating agent. These results showed that glucocorticoid hormone recovered TPA-induced reduction of taurine transporter activity and that IM recovered CsA-induced reduction of the transporter activity by increasing intracellular free $Ca^{++}$ concentration.n.

• Korean Journal of Food Science and Technology
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• v.23 no.2
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• pp.188-191
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• 1991

A study was carried out to investigate the control effect of sodium and potassium phosphates, sodium citrate and three different salts mixtures on kimchi fermentation when they were added into half-fermented kimchi in the concentration range of $0.001{\sim}0.01\;M$. The salts mixtures added were sodium phosphates mixture(CA-A), addition of $NaNO_2$, Ca-EDTA and BHA to CA-A(CA-B) and substitution of BHA with sodium citrate in CA-B. The results showed that sodium phosphates and sodium citrate significantly inhibited the kimchi fermentation while potassium phosphate had little effect. The order of control effect was $Na_3PO_4-Na_2HPO_4-sodium\;citrate-NaH_2PO_4-K_2HPO_4-KH_2PO_4$. Among the salts mixtures, CA-A showed the most reducing effect in the fermentation rate followed by CA-C and CA-A. The mixture of CA-C could extend the time of holding pH $4.2{\sim}4.4$ by approximately 6 times at $4{\sim}25^{\circ}C$ when it was compared to control. The microbial growth study of total and Leuconostoc mesenteroides also showed a very significant decrease in their numbers.

• Journal of the Korean Society of Food Science and Nutrition
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• v.14 no.1
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• pp.82-87
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• 1985

Some biological activities showed as ATPase activity of myofibril, actomyosin and myosin extracted from chicken leg and pectoral skeletal muscle were investigated. The $Mg^{+2}$-ATPase activity at 0.05 M KCl were 0.82, 0.38 and 0.11 ${\mu}mole$ Pi/mg protein/min. in actomyosin, myofibril and myosin from pectoral muscle while 0.71, 0.32 and 0.08 ${\mu}mole$ Pi/mg protein/min. in actomyosin, myofibril and myosin from leg muscle. EDTA-ATPase activity at 0.6M KCl were 0.80, 0.42 and 0.40 ${\mu}mole$ Pi/mg protein/min. in actomyosin, myofibril and myosin from pectoral muscle. In case of leg muscle, that activity was noted as 0.69, 0.33 and 0.28 ${\mu}mole$e Pi/mg protein/min in proteins. ATPase activity of myosin from leg and pectoral muscle were inhibited in 10% at a higher concentration of $Mg^{+2}$ than molar concentration of EDTA, and the ATPase activity was increased to 400% compared with control at $10^{-3}M$ of $Ca^{+2}$. Actomyosin from pectoral muscle was solubilized at 0.1 M KCl above and that from leg muscle was solubilized at 0.15 M KCl above. In case of myosin, pectoral muscle was solubilized at 0.25M KCI above and leg muscle was solubilized at 0.30M KCl above.

• Korean Journal of Agricultural Science
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• v.32 no.1
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• pp.71-80
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• 2005

Phytic acid chelates excellently the metallic ions and the positive ions, especially has high affinity with $Fe^{2+}$ and $Ca^{2+}$. Merits of phytic acid can be taked in easily, edibile and harmless to body, so it was investigated that phytic acid can be substituted for EDTA in this study. 1. The Intensificative effect of chelating agent and disinfective osmotic shock of Vibrio vulnificus The number of initial existent fungi measured $1.7{\times}10^6$. The percentages of the survival fungi against the osmotic shock by distillated water were calculated at 1 minute, 3 minute and 5 minute after inoculation. The percentages of the survival fungi in $Mg^{2+}$ were 92.5%, 91.8% and 79.8% at each time, the average percentage was 88%. Also the sudden extinction was observed around 1 minute after inoculation and the survival fungi were not observed from 3 through 5 minute in spite of repeated experimentation. 2. Influence of Vibrio vulnificus on the survival of the mice. The first mouse started to die in 180 minute after inoculation in case that the inoculating number was $2.3{\times}10^7cfu/ml$. All died within 4.5 hour. The average of survival time was 226 minute. The first mouse started to die in 228 minute after inoculation in case that the inoculating number was $0.8{\times}10^6cfu/ml$. All died within 5 hour. The average of survival time was 300 minute and the survival time was 1.3 times high. The tendencies of death in two cases were similar, but the fatal rate were largely dependent on inoculating number.

• The Korean Journal of Pharmacology
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• v.32 no.3
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• pp.425-431
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• 1996

Human neutrophil elastases (HNElastase, EC 3.4.21.37), a causative factor of inflammatory diseases, are regulated by plasma proteinase inhibitors, alpha-proteinase inhibitor and ${\alpha}_2-macroglobulin$. Under certain pathological conditions, however, released enzymes or abnormal function of inhibitors may cause various inflammatory disease. NSAIDs have been clinically applied for treatment of inflammatory diseases. Inhibition of cyclooxygenase is a known mechanism of action of NSAIDs in the treatment of inflammatory disease. In in vitro experiments, HNElastase was inhibited by naproxen, phenylbutazone, and oxyphenbutazone, but ibuprofen, ketoprofen, aspirin, salicylic acid, and tolmetin did not inhibit elastase. HNElastase was also inhibited by chelating agents, EDTA & EGTA, and tetracyclines. Removal of divalent metal ions by EDTA caused inhibition of elastase, and reconstitution of the metal ions recovered the enzyme activity to a certain level. Frequencies and contours in the Raman spectra of various conditions of human neutrophil elastase undergo drastic changes upon partial removal and/or reconstitution of calcium and zinc ions. The metal ion content dependent activities and change of the contour of the Raman spectrogram suggest us that the mechanism of action of a chelator or chelator-like agents on neutrophil elastase may be related to the conformational change at/or near the active site, especially -C=O radical or -COOH radical.

• Journal of the Korean Society of Clothing and Textiles
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• v.7 no.2
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• pp.19-25
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• 1983

The purpose of this study was to investigate the effects of laundry variables and additives on the removal of deposited calcium on the cotton fabric. Samples of calcium deposited fabric was made by treating fabric with $CaC1_2$ and $Na_2CO_3$ solution subsequently. The experimental variables were: 1) NaOH concentration ($0.0001\%$, $0.0005\%$, $0.001\%$, $0.005\%$, $0.01\%$) 2) Alkaline builders(sodium carbonate, sodium meta silicate) 3) Sequestering agents(STPP and EDTA concentration: $0.02\%$, $0.04\%$, $0.06\%$, $0.08\%$, $0.1\%$, $0.15\%$, $0.2\%$) 4) Temperatures($25\pm1^{\circ}C$, $40\pm1^{\circ}C$, $60\pm1^{\circ}C$) 5) Edge-abrasion to the removal of deposited calcium on the cotton fabric. The fabric was washed for 15 minutes in a washing machine(Model: Gold Star WP-3007) or Launder-0-meter(40$\~$45 r.p.m., Toyo Rika Instrument Inc.) and rinsed 3 times per every rinsing time. The amount of calcium deposits on the fabrics was determined by EDTA-back titration methods and edge-abrasion was evaluated by ASTM D 3886 method. The results of this study were as follows: 1) pH of surfactant solution(NaOH concentration) did not influence on the removal of deposited calcium on the cotton fabric. 2) Added alkaline builders did not influence on the removal of deposited calcium on the cotton fabric. 3) It was shown that STPP and EDTA were effective to remove deposited calcium. The removal of deposited calcium on the cotton fabric was proportionally increased with increasing concentration of STPP and EDTA. At high concentration, however, the rate was rather decreased with increasing concentration. 4) The temperature of washing solution did not influence on the removal of dedosited calcium on the cotton fabric. 5) As the removal of deposited calcium on the cotton fabric was increased, the rate of edge-abrasion of the fabric was gradually increased.

• Korean Journal of Microbiology
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• v.35 no.4
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• pp.315-321
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• 1999

This study has been performed to deveope a yeast strain having high ${\alpha}$-amylase production ability using nuclear transfer method. Hybrids formed between the strains of Saccharomyces fiburigera KCTC 7393 and Saccharomyces cerevisiae KCTC 7049 (tyr-, ura-)were obtained by nuclear transfer technique. Nuclei isolated from the wild type S. fiburigera strain were transfered into auxotrophic mutants S. cerevisiae and selected the hybrids showing an increased starch degrading capability were selected (MN-16). This transformant grew best and produced maximal ${\alpha}$-amylase activity on the medium containing 2% (V/V) soluble starch. ${\alpha}$-Amylase from MN-16 was purified electrophoretically homogenety and its properties were investigated. The enzyme was purified about 10.6 fold with an overall yield 9.7% from the culture medium by ammonium sulfate fractionation. DEAE-Sephacel column chromatography, and Sephacryl S-200 column chromatography. The purified enzyme showed a single band on SDS-polyacrylamide gel electrophoresis. The molecular weight of the ${\alpha}$-amylase was estimated to be 53,000 daltons by SDS-PAGE and by gel permeation chromatography on Sephacryl S-200. The purified enzyme showed the maximum activity at pH 5.5 and 40${\circ}C$. The km value for soluble starch was 2.5㎎/㎖. The enzyme activity increased in the presence of $Ca^{2+}, Co^{2+}, EDTA, Mg^{2+}, Mn^{2+}, Zn^{2+}$, but inhibited by $Cu^{2+}, Fe^{2+}$, and $Ni^{2+}$

• Microbiology and Biotechnology Letters
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• v.16 no.2
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• pp.142-149
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• 1988

This experiment was carried out to obtain a hybrid with potent ethanol fermenting ability, by means of protoplast fusion between a thermophilic strain (D-71) of Saccharomyces cerevisiae and an osmotolerant strain (SR-S) of Zygosaccharomyces rouxii. The conditions for formation of protoplasts from both strains and for their fusion and regeneration were studied. Favorable conditions for formation of protoplasts from Saccharomyces cerevisiae D-71 were : treatment of the cells at late-exponential phase with 2-mercaptoethanol (l% v/v) for 10 minutes in the presence of 0.5M sorbitol, then incubation for 60 minutes in the set medium containing Zymolyase-20T (4mg/$m\ell$) ; and from Zygosaccharomyces rouxii SR-S were : treatment of the cells at mid-exponential phase with 2-mercaptoethanol (1% v/v) for 10 minutes in the presence of 0.5M or 1M mannitol, then incubation for 120 minutes in the set medium containing Zymolyase-20T(4mg/$m\ell$). The protoplasts of parental cells were fused in the presence of 20mM CaCl$_2$, 0.5M sorbitol and 40% of polyethyleneglycol (M.W 4000), then fusants obtained were selected as regenerated colonies which embedded and grown in the minimal medium containing 3% of agar. The frequencies of fusant formation were 1.2$\times$10$^{-6}$ to 9.1$\times$10$^{-6}$ for the regenerated protoplast.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.10
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• pp.1466-1472
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• 2012

A bacterial isolate derived from soil samples near a cattle farm was found to display extracellular phytase activity. Based on 16S rRNA sequence analysis, the strain was named Bacillus sp. T4. The optimum temperature for the phytase activity toward magnesium phytate (Mg-$InsP_6$) was $40^{\circ}C$ without 5 mM $Ca^{2+}$ and $50^{\circ}C$ with 5 mM $Ca^{2+}$. T4 phytase had a characteristic bi-hump two pH optima of 6.0 to 6.5 and 7.4 for Mg-$InsP_6$. The enzyme showed higher specificity for Mg-$InsP_6$ than sodium phytate (Na-$InsP_6$). Its activity was fairly inhibited by EDTA, $Cu^{2+}$, $Mn^{2+}$, $Co^{2+}$, $Ba^{2+}$ and $Zn^{2+}$. T4 phytase may have great potential for use as an eco-friendly feed additive to enhance the nutritive quality of phytate and reduce phosphorus pollution.