• Microbiology and Biotechnology Letters
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• v.19 no.3
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• pp.209-214
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• 1991

Purification of the bacteriocin from Lactococcus sp. 1112-1 was achieved by successive column chromatography on CM-Sephadex C-25 and Sephadex G-50, starting from cell disruption broth. 16.2% of the initial activity was recovered after this purification step and it was shown 123-fold increase in purification. Purified bacteriocin was shown a single band on SDS-polyacrylamide gel electrophoresis. This substance was rather stable at heat treatment and alkaline pH relatively. The residual antimicrobial activity was 38% when the bacteriocin was treated by heat at $100^{\circ}C$ for 60 min. And 23% of the activity remained at pH 8.0 after standing for 48 hr. The amino acid composition of purified bacteriocin was made up 26 residues.

• Journal of the Korean Wood Science and Technology
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• v.30 no.3
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• pp.12-17
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• 2002

The extracellular invertase (EC 3.2.1.26) (Candida utilis) preparation was obtained from the liquid medium after desalting and freeze drying. This prepared enzyme was used for the comparative purification on 4 activated matrices by liquid column affinity chromatography method. In this method there were used controlled porous glass (CPG) silanized covalently activated by keratin, silanized silica gel and silica gel covalently covered by keratin. It was found that the invertase purification process was better using both CPG matrices (silanized CPG and keratin activated CPG) than these with two silica gel supports. Also the elution coefficient of the invertase from the two CPG columns was about 93 to 94%. Two silica gel supports found to be superior in terms of purification efficiency. The invertase purification process was confirmed by PAGE electrophoresis.

• Applied Biological Chemistry
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• v.21 no.2
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• pp.112-118
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• 1978

Xanthine oxidase from bovine thyroid glands was purified to apparent homogeneity when judged by analytical disc gel electrophoresis. The purification procedures include pancreatin digestion, butanol extraction, ammonium sulfate precipitation, calcium phosphate gel adsorption, ultrafiltration, calcium phosphate gel-cellulose column chromatography, gel filtration, preparative Sephadex G-25 column electrophoresis, and preparative polyacrylamide gel electrophoresis. The enzyme was enriched 1,000-fold. However, its specific activity was markedly low as compared with highly purified milk enzyme. Thyroidal xanthine oxidase exhibited a low specificity for substrates and electron acceptors. The kinetic properties of thyroid xanthine oxidase were found to be similar to those of the milk enzyme on the basis of Michaelis constants for common substrates.

• Korean Chemical Engineering Research
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• v.56 no.5
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• pp.738-751
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• 2018

The effect of the reaction temperature and reaction time on the thermal oxidative purification quality of detonation nanodiamond (NDsoot) was investigated in a gas-solid fluidized bed reactor of a $0.10m-ID{\times}1.0m$-high stainless steel column with zirconia beads ($d_{SV}=99.2{\mu}m$). The carbon conversion increased with increasing the reaction temperature; however, when the reaction temperature was greater than 773 K, the carbon conversion did not increase. The content of $sp^3$-hybridized carbon at the reaction temperature of 703 K barely changed when the reaction time was more than 30 minutes, but at 773 K, the content decreased as preferred. At 703 K, the purification quality increased with the increasing reaction time; however, at 773 K, the purification quality increased up to 30 minutes and then decreased rapidly.

• Applied Biological Chemistry
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• v.51 no.3
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• pp.153-158
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• 2008

An actinomycetes F-97 producing tyrosinase inhibitor was isolated from soil samples. Isolation and purification of tyrosinase inhibitor produced by F-97 was performed as follows: IRC-120 ($NH_4^+$ type) column chromatography, silica gel column chromatography, $C_{18}$ column chromatography and Sephadex LH-20 column chromatography were used successively after the centrifuged supernatant was adjusted to pH 4.0. To identify the purity of the inhibitor, octadecylsilyl(ODS) HPLC was carried out with 5% methanol as a mobile phase. Finally, the purification yield of a tyrosinase inhibitor was 5.24%. The inhibitor was very soluble in water, methanol and ethanol but insoluble in acetone, butanol, ethylacetate and chloroform. The ${\lambda}_{max}$ value of this inhibitor in water was 194nm under UV light. The biochemical test of the inhibitor was positive in Molish, Benedict, cone. $H_2SO_4$, and $KMnO_4$ tests but negative in iodine, ninhydrin, Million, Sakaguchi, xanthoproteic and Emerson tests. The tyrosinase inhibitor was stable against heat treatment of $100^{\circ}C$ for 50 minutes and pH $4{\sim}9$. The $IC_{50}$ value of this inhibitor was $19.2{\mu}g/ml$ for mushroom tyrosinase. In $1,000{\mu}g/ml$ inhibitor concentration, inhibition zone was 27 mm for Streptomyces bikiniensis NRRL B-1049. The inhibition of F-97 against mushroom tyrosinase was competitive with tyrosine.

• Analytical Science and Technology
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• v.21 no.4
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• pp.338-343
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• 2008

Extraction of DNA from skeletal material is of great importance in the identification of human remains, but is particularly difficult because the high amount of microbial DNA was often co-extracted with human bone DNA. We found that a phenol/chloroform extraction, followed by ultrafiltration, and cleanup by via the $QIAquick^{(R)}$ PCR purification kit yields higher amounts of human genomic DNA compared with extraction by the column affinity $method^{(R)}$ alone. Ultrafiltration extraction of human DNA from ten exhumed bone samples yielded $0.041-1.120ng/{\mu}L$ DNA (mean = $0.498ng/{\mu}L$ DNA), and purification using the column affinity resulted in $0.016-0.064ng/{\mu}L$ DNA (mean = $0.034ng/{\mu}L$ DNA). Although the STR genotyping by the column affinity method was partially successful, all DNA samples by the ultrafiltration method produced full profiles from the multiplex PCR. The efficiency of STR genotyping was in accordance with the amounts of the human DNA extracted.

• Journal of Microbiology and Biotechnology
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• v.18 no.10
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• pp.1648-1654
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• 2008

Superoxide dismutase (SOD) removes damaging reactive oxygen species from the cellular environment by catalyzing the dismutation of two superoxide radicals to hydrogen peroxide and oxygen. Extracellular superoxide dismutase (EC-SOD) is a tetramer and is present in the extracellular space and to a lesser extent in the extracellular fluids. Increasing therapeutic applications for recombinant human extracellular superoxide dismutase (rEC-SOD) has broadened interest in optimizing methods for its purification, with a native conformation of tetramer. We describe a solid phase refolding procedure that combines immobilized metal affinity chromatography (IMAC) and gel filtration chromatography in the purification of rEC-SOD from Escherichia coli. The purified rEC-SOD tetramer from the $Ni^{2+}$-column chromatography is refolded in Tris buffer. This method yields greater than 90% of the tetramer form. Greater than 99% purity is achieved with further purification over a Superose 12PC 3.2/30 column to obtain the tetramer and specific activities as determined via DCFHDA assay. The improved yield of rEC-SOD in a simple chromatographic purification procedure promises to enhance the development and therapeutic application of this biologically potent molecule.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.26 no.6
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• pp.529-537
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• 1993

Several methods were examined for purification of docosahexaenoic acid (DHA, 22:6n-3) from skipjack Euthynnus pelamis orbital tissue oil, a marine by-product, and a modified method for isolation of a high purity DHA was proposed. Skipjack orbital tissue contained $55.4\%$ of total lipid(TL), and DHA accounted for $23.7\%$ of the TL. Application of low-temperature crystallization and urea inclusion compound methods to the orbital fatty acid mixture resulted in increases of DHA concentrations to approximately $46\%\;and\;61\%$, respectively. These methods were suitable for large production of DHA with relative low purity because of the simple purification procedure. DHA of approximately $74\%$ in purity was obtained by silver nitrate aqueous solution method, but the method gave a very low recovery($<10\%$). Silver nitrate-impregnated silica column chromatography was suitable for purification of a high purity DHA(purity, $>98\%$ and recovery, $>90\%$) A modified method, silver nitrate-impregnated silica column chromatography combined with low-temperature crystallization(two step purication method) was proposed as the most effective method to obtain DHA with high purity($99.9\%$) from the skipjack orbital oil.

• Journal of Life Science
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• v.14 no.2
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• pp.225-228
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• 2004

Natamycin, produced by Streptomyces natalensis ATCC27448, is a polyene macrolide antibiotic, is widely used in the food industry in order to prevent mould contamination. This study carried out to develop an efficient purification process of natamycin from fermentation broth. The stability of natamycin in fermentation broth during storage period was investigated at 4$^{\circ}C$ and room temperature. After the storage of fermentation broth for 14 days at 4$^{\circ}C$, residual activity of natamycin was about 80% but decreased by 27% at room temperature. As solvent to extract natamycin from fermentation broth, methanol was the most efficient. A developed purification procedure includes methanol extraction and Diaion HP-20 column chromatography. Approximately 2.9 g of natamycin was obtained with a final yield of 69.1% and purity of 96.6% from 1.8 l of fermentation broth by this developed purification procedure.

• BMB Reports
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• v.29 no.4
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• pp.335-342
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• 1996

Porcine liver cysteinesulfinic acid decarboxylase was purified approximately 460-fold by means of ammonium sulfate fractionation and sequential column chromatographic separation with Sephadex G-100, DEAE-cellulose and hydroxylapatite. The enzyme has a flat pH profile with maximum activity occurring between pH 6.0 and 7.6. Pyridoxal 5'-phosphate must be present in all buffers used for purification procedures in order to stabilize the enzyme. Addition of sulfhydryl reagents such as 2-mercaptoethanol are also necessary to maintain maximum enzyme activity throughout purification. The absorption spectrum shows that cysteinesulfinic acid decarboxylase is a pyridoxal 5' -phosphate-containing protein. The major absorption is at 280 nm with two smaller absorption regions, one at 425 nm which is ascribed to a Schiffs base between pyridoxal phosphate and protein, and another at 325 nm which is thought to be due to the interaction of 2-mercaptoethanol with the Schiffs base. A number of divalent cations tested did not affect enzyme activity with the exception of mercury, copper, and zinc which are inhibitory. The partially purified enzyme has an apparent $K_m$ of 0.94 mM for cysteinesulfinate. Cysteic acid is a competitive inhibitor of the enzyme with a $K_i$ of 1.32 mM. The molecular weight of the enzyme was estimated to be about 79,600 by using Sephadex G-200 column chromatography.