• Journal of the Korean Chemical Society
• /
• v.28 no.5
• /
• pp.309-314
• /
• 1984

The species and equilibria of uranium and vanadium have been investigated in the various concentration of perchloric, hydrochloric and sulfuric acid by anion exchange chromatography. In the concentration range of $0.01\;{\sim}\;0.5M$ hydrochloric and $0.01\;{\sim}\;0.5M$ perchloric acid, uranium seems to be $UO_2^{2+}$species and in higher concentration than 0.5M hydrochloric acid $UO_2^{2+}$seems to form the chloride complex ion as $UO_2Cl^+$, $UO_2Cl_2$, $UO_2Cl_3^-$ and $UO_2Cl_4^{2-}$ according to the increase of the hydrochloric acid concentration. In the dilute(0.01N) sulfuric acid the adsorbability of uranium on anion exchange resin is strong and then decreases with increasing the sulfuric acid concentration. From this result we conclude that $UO_26{2+}$ formed the complex ion as $UO_2(SO_4)_2^{2-}$. In the perchloric acid of $0.01\;{\sim}\;0.5N$ concentration the existing equilibrium of vanadium and its constant calculated at $20^{\circ}C$ is $1.9{\times}108$ for $H_2V_{10}O_{28}^{4-}$ + $14H^+$ = $10VO_2^+ + 8H_2O$. The elution behaviors of vanadium in the hydrochloric and sulfuric acid are smiliar to those in perchloric acid.

• Applied Biological Chemistry
• /
• v.41 no.1
• /
• pp.6-12
• /
• 1998

To extract insoluble proteins and to improve functional properties of abolished proteins, a protease producing Aspergillus sp. MS-18 was isolated from soil. The enzyme was purified and its enzymological characteristics were investigated. It was found that production of protease reached to the maximum when the wheat brae medium containing, 3% arabinose, 0.5% polypepton, 0.1% $(NH_4)_2SO_4$ and 0.2% magnesium chloride was cultured for 3 days. Protease was purified 16.9 folds after ion exchange chromatography and gel filtration and the specific activity was 340.4 unit/mg. Purified enzyme was confirmed as a single band by the polyacrylamide gel electrophoresis. The molecular weight of protease was estimated to be 30,000. Crystalization form of purified protease was a stick shape with rounding edges. The optimum pH and temperature for the protease activity were 9.0 and $60^{\circ}C$, respectively. The enzyme was stable in pH 7.0-12.0 at $50^{\circ}C$. The activity of purified enzyme was inhibited by $Hg^{2+}$, $Cu^{2+}$, $Zn^{2+}$ and $Pb^{2+}$, whereas it was activited by $Na^+$, $Mg^{2+}$ and $Mn^{2+}$. The activity of the protease was inhibited by the treatment with ethylenediaminetetraacetic acid and phenylmethane sulfonyl fluoride. The result suggests that the purified enzyme is a serine protease with metal ion at active site. Km and Vmax of purified protease were $29.33\;{\mu}mole/L$ and $5.13\;{\mu}g/min$, respectively.

• The Korean Journal of Pharmacology
• /
• v.10 no.2
• /
• pp.31-37
• /
• 1974

Citrus unshiu Marc., a family of Rutaceae, has been used as antitussives, antiemetics, expectorants, spasmolytics, stomachics and digestants in oriental medicine. But there is few reports on the pharmacological effects of Citrus unshiu pericarpium. Present experiment was performed to observe the effects of the water extract of Citrus unshiu on isolated auricle and intestine in rabbits and blood pressure in rats and also to study on the active component on the cardiovascular system by means of thin layer chlomatography and spectrophotometer. 1. The water extract showed positive inotropic and positive chronotropic effect in the concentration of $1{\times}10^{-4}g/ml$, $5{\times}10^{-4}g/ml$, and $1{\times}10^{-3}g/ml$. 2. The motility of isolated rabbit intestine was significantly depressed by the water extract. 3. The blood pressure was markedly increased by the water extract. 4. The blood vessel of rabbit ear was constricted by perfusion of the ether extract. 5. The purified alkaloid of Citri pericarpium by ion exchange resin showed the same Rf value with l-synephrine on thin layer chromatography. 6. The purified alkaloid of Citri pericarpium by thin layer chromatography showed the same absorption curve with l-synephrine in the ultraviolet scanning.

• The Journal of the Korean Society for Microbiology
• /
• v.34 no.5
• /
• pp.461-470
• /
• 1999

Growth under conditions of iron-restriction and the production of siderophore was examined in Vibrio mimicus ATCC 33653. This strain grew and multiplied in the presence of the high-affinity iron chelators ethylenediamine-di (o-hydroxyphenylacetic acid). Chrome azurol S (CAS) agar and solution were used to detect the production of siderophore under these condition. Siderophore could be detected in the iron-restricted culture supernatants. The siderophore was extracted from iron-restricted culture supernatants by phenol-chloroform-ether method and purified by Dowex ion-exchange and Sephadex G-25 gel filtracton chromatography. The purified siderophore was confirmed by paper chromatography and HPLC. The Purified siderophore enhanced the growth of V. mimicus when the bacterium was grown in iron limited medium. Injection of both the siderohore and the bacteria to mice resulted in more rapid death than that of the only bacteria. However, the siderophore did not show lethality to mice and any toxicity to cell line like HeLa and U937.

• Journal of Microbiology and Biotechnology
• /
• v.9 no.4
• /
• pp.469-474
• /
• 1999

Thermoactinomyces sp. E79 produced two types of thermostable alkaline proteases extracellularly. A minor protease was separated from a major protease by using DEAE-column chromatography. This enzyme was purified to homogeneity by ammonium sulfate and DEAE-Sepharose ion-exchange chromatography. The purified minor protease showed different biochemical properties compared to the major protease. The molecular mass of the purified enzyme was estimated by SDS-PAGE to be 36 kDa. Its optimum temperature and pH for proteolytic activity against Hammarsten casein were $70^{\circ}C$ and 9.0, respectively. The enzyme was stable up to$75^{\circ}C$ and in an alkaline pH range of 9.0-11.0. The enzyme was inhibited by phenylmethylsulfonyl fluoride (PMSF) and $Hg^{2+}, indicating that the enzyme may be a cysteine-dependent serine protease. In addition, the enzyme cleaved the endoproteinase substrate, succinyl-Ala-Ala-Pro-Phe-p- nitroanilide, and the $K_m$ value for the substrate was 1.2 mM.

• Microbiology and Biotechnology Letters
• /
• v.24 no.4
• /
• pp.478-484
• /
• 1996

Recombinant human epidermal growth factor (rhEGF) was produced by E. coli BL21 harboring a plasmid pYHB101. The maximum production was 68.7 mg/l when the E. coli strain was cultured at 25$\circ$C for 48 hours in the modified MBL medium containing 10 g/l glucose with 1 mM IPTG induction at 2 hours after inoculation. The rhEGF was purified upto 267 folds by Amberlite XAD- 7 chromatography, ultrafiltration, and DEAE Sepharose fast flow ion exchange chromatography with an overall yield of 66.6%. The purified rhEGF was further separated into two fractions by HPLC. The N-terminal amino acid sequence of the second fraction was Asn-Ser-Asp-Ser-Glu-Cys-Pro-Leu-Ser-His. The effect of rhEGF on the DNA synthesis was examined using in vitro biological assay based on the incorporation of 5'-bromo-2'- deoxy-uridine (BrdU). The purified rhEGF shows no difference with natural human epidermal growth factor (nhEGF) in N-terminal amino acids residues and biological activity. From the results, we concluded that rhEGF produced from E. coli harboring the plasmid pYHB101 was apparently the same as nhEGF.

• Journal of Veterinary Clinics
• /
• v.16 no.1
• /
• pp.31-36
• /
• 1999

The aim of this study is to determine the phagocytosis-promoting factor(s) for feline peripheral blood polymorphonuclear cells (PMN) by culture supernatant from mono-nuclear cells (MNC) treated with egg white derivatives (EWD). The phagocytic activity of PMN was analyzed by a flow cytometry system. The EWD did not show direct effect on the phagocytic response of PMN. The phagocytic activity of PMN was enhanced by culture supernatant from MNC but not PMN treated with EWD. Therefore, it was suggested that the enhanced phagocytic activity of feline PMN could be mediated by humoral factor(s) released from MNC treated with EWD. Thus, the phagocytosis-promoting factor(s) in supernatant fraction from MNC culture treated with EWD were isolated by reverse phase high pressure liquid chromatography. The resulting supernatant fraction on 29.02 minutes of retention time showed high phagocytic activity of PMN. The molecular weight of this supernatant fraction was 16 to 18 kDa when analyzed by capillary electrophoresis. The isoelectric point was pH 5.76 when assessed by ion-exchange chromatography. These results suggest that EWD stimulates feline MNC to elaborate a phagocytosis-promoting factor, 16 to 18 kDa of molecular weight, which could be an important mediator for the enhancement of phagocytic activity of feline peripheral blood phagocytes. Further study will be needed to elucidate this phagocytic factor.

• BMB Reports
• /
• v.32 no.6
• /
• pp.605-609
• /
• 1999

Thioltransferase, also called glutaredoxin, is a general GSH-disulfide reductase of importance for redox regulation. Previously, the protein thioltransferase, now called S-type thioltransferase, was purified and characterized from Arabidopsis thaliana seed. In the present study, a second thioltransferase, called L-type thioltransferase, was purified to homogeneity from Arabidopsis thaliana leaves. The purification procedures included DEAE-cellulose ion-exchange chromatography, Sephadex G-50 gel filtration, and glutathione-agarose affinity chromatography. The purified enzyme was confirmed to show a unique band on SDS-PAGE and its molecular weight was estimated to be 26.6 kDa, which appeared to be atypical compared with those of most other thioltransferase. It could utilize 2-hydroxyethyl disulfide, S-sulfocysteine, and insulin as substrates, and also contained dehydroascorbate reductase activity. Its optimum pH was 8.5 and its activity was greatly activated by L-cysteine. When it was kept for 30 min, it appeared to be very stable up to $70^{\circ}C$. It was activated by $MgCl_2$ and, on the contrary, inhibited by $ZnCl_2$, $MnCl_2$, and $AlCl_3$.

• BMB Reports
• /
• v.28 no.2
• /
• pp.100-106
• /
• 1995

S-adenosylmethionine (SAM) synthetase was purified to homogeneity from soybean (Glycine max) axes. The enzyme was purified 216-fold with a 1.5% yield by ammonium sulfate fractionation, acetone fractionation, ion exchange chromatography with DEAE-sephacel, gel filtration with Sephacryl S-300, and afffinity chromatography with ATP-agarose. The enzyme activity reached a maximum 3 days after germination. SAM synthetase had a subunit molecular weight of 57,000 daltons from a silver stained single band on SDS-PAGE. The molecular weight of the enzyme was 110,000 daltons from Sephacryl S-300 gel filtration. The enzyme was composed of two identical subunits. The $K_m$ values of the enzyme for L-methionine and ATP were 1.81 and 1.53 mM, respectively. The enzymatic activity was not affected by polyamines, agmatine, or SAM analogues, but was inhibited by SAM. The inhibition pattern was showed non-competitive for L-methionine and uncompetitive for ATP. The activity of SAM synthetase was inhibited by thiol-blocking reagents. The enzyme was induced by treatment with $10^{-3}$ M putrescine at germination. Experimental data revealed a possible novel regulation mechanism of polyamine biosynthesis through several endogenous intermediates.

• Journal of Nutrition and Health
• /
• v.16 no.2
• /
• pp.115-124
• /
• 1983

The biochemical nature of the protein constituents of six year old fresh Panax ginseng root was studied. Total protein constituents were extracted with phosphate buffer of pH 7.4, ionic strength of 0.1 and fractionated by ultrafiltration using four different membranes which cut down the materials of molecular weight of 500, 1,000, 5,000 and 10,000, respectively. Each fraction was subjected to ion exchange chromatography using DEAE - cellulose to isolate component proteins. The protein fraction larger than molecular weight of 10,000 was refractionated by the method of ammonium sulfate precipitation. The electrophoresis of the refractionated protein constituents was performed. The amino acid composition of the protein constituents was determined by gas- liquid chromatography. From the results, it could be summarized that eleven different protein constituents smaller than molecular weight of 10,000 were isolated from the fresh Panax ginseng root. At least eleven different protein constituents larger than molecular weight of 10,000 were identified from the electrophoretic patterns. These protein costituents seem to be compounded of all or some of five different subunits.