• 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.

• Fisheries and Aquatic Sciences
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• v.15 no.3
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• pp.191-195
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• 2012

We report here the use of high-speed countercurrent chromatography (HSCCC) in the preparative isolation and purification of the bioactive component, fucosterol, from Pelvetia siliquosa. A crude extract was obtained by ultrasonic extraction of powdered P. siliquosa using methylene chloride and was then subjected to separation and purification by HSCCC, coupled with evaporative light-scattering detection. Preparative HSCCC was performed successfully using a two-phase solvent system, n-heptane:methanol (3:2, v/v), to obtain 10.96 mg fucosterol with 96.8% purity from 50 mg of crude extract; the recovery rate was approximately 90.5%.

• 한국약용작물학회:학술대회논문집
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• 2007.11a
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• pp.292.2-292.2
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• 2007

• YAKHAK HOEJI
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• v.41 no.5
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• pp.638-646
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• 1997

The steroid 9${alpha}$-hydroxylase activity has been detected in cytosol fraction, $100,00{\times}g$ supernatant of cell free extract of Mycobacterium fortuitum. The activity was not linear with protein concentration in the assay suggesting 9${alpha}$-hydroxylase is a multicomponent enzyme. The 9${alpha}$-hydroxylase system was partially purified through fractional saturation of ammonium sulfate, strong anion exchange (Mono Q) column chromatography, gel filtration (Superose 12) column chromatography, and testosterone affinity gel chromatography. Ammonium sulfate 50~60% saturated fraction of the cytosol gave 9${alpha}$-hydroxylase activity. For further purification, the half-saturated ammonium sulfate fraction was applied to Mono Q, Superose 12, or affinity gel column. The purification factors of 9${alpha}$-hydroxylase containing fraction after Mono Q, Superose 12, and affinity gel chromatography was 13, 11, and 17 respectively.

• KSBB Journal
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• v.9 no.4
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• pp.385-394
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• 1994

Large scale purification to get antifungal antibiotic KRF-001 of 90% purity, was investigated using preparative HPLC. Crude KRF-001 was purified by XAD-7 adsorption chromatography, acid precipitation and microfiltration. Microfiltration was the most effective isolation method of crude KRF-001. The purification methods using C18 chromatography was convenient compared with the conventional methods. Delta PAK C18 column and Bonda PAK C18 column were adapted large scale purification of KRF-001. Gradient system of prep HPLC using Delta PAK C18 column was more effective. With these conditions, final recovery of KRF-001 yielded 77%.

• Journal of Ginseng Research
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• v.13 no.2
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• pp.254-259
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• 1989

A radioprotective ginseng protein fraction was obtained from Korean white ginseng powder by the following isolation and purification procedures: Tris-HCI buffer extraction, 70% ammonium sulfate fractionation, CM-rellulosr column chromatography, heat inactivation and Sephadex G-75 column chromatography. This fraction was further purified by Sepharose 4B and Sephadex G-150 column chromatographies. Three fractions obtained were subjected to Native-PAGE and SDS-PAGE using gradient gels and the silver staining method. Molecular weights of the native proteins and their subunits were estimated.

• Journal of Microbiology and Biotechnology
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• v.22 no.7
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• pp.930-938
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• 2012

High levels of xylanase activity (143.98 IU/ml) produced by the newly isolated Paenibacillus campinasensis G1-1 were detected when it was cultivated in a synthetic medium. A thermostable xylanase, designated XynG1-1, from P. campinasensis G1-1 was purified to homogeneity by Octyl-Sepharose hydrophobic-interaction chromatography, Sephadex G75 gel-filter chromatography, and Q-Sepharose ion-exchange chromatography, consecutively. By multistep purification, the specific activity of XynG1-1 was up to 1,865.5 IU/mg with a 9.1-fold purification. The molecular mass of purified XynG1-1 was about 41.3 kDa as estimated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Sequence analysis revealed that XynG1-1 containing 377 amino acids encoded by 1,134 bp genomic sequences of P. campinasensis G1-1 shared 96% homology with XylX from Paenibacillus campinasensis BL11 and 77%~78% homology with xylanases from Bacillus sp. YA-335 and Bacillus sp. 41M-1, respectively. The activity of XynG1-1 was stimulated by $Ca^{2+}$, $Ba^{2+}$, DTT, and ${\beta}$-mercaptoethanol, but was inhibited by $Ni^{2+}$, $Fe^{2+}$, $Fe^{3+}$, $Zn^{2+}$, SDS, and EDTA. The purified XynG1-1 displayed a greater affinity for birchwood xylan, with an optimal temperature of $60^{\circ}C$ and an optimal pH of 7.5. The fact that XynG1-1 is cellulose-free, thermostable (stability at high temperature of $70^{\circ}C{\sim}80^{\circ}C$), and active over a wide pH range (pH 5.0~9.0) suggests that the enzyme is potentially valuable for various industrial applications, especially for pulp bleaching pretreatment.

• Proceedings of the Korean Vacuum Society Conference
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• 2013.02a
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• pp.87-87
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• 2013

Sample concentration and purification processes are essential in the bio-analytical and pharmaceutical fields because most bio samples or media are extremely sophisticated. To concentrate and purify specific substances, passive membrane type filters have been utilized, which is driven by size or charge differences between target and others. The traditional and representative method to identify nucleic acid sequences in the complex biosample is gel electrophoresis, which has been worked by size and net charge of molecules. The adsorption phenomena have been also utilized to concentrate and purify biomolecules. This adsorption of biomolecule can be controlled under specific salts and surfaces as well as surface area. To utilize the differences of physical properties of molecules or bio-targets such as virus, bacteria, and cells, the nanotechnologies can be introduced in target concentration, purification, and isolation processes. In here, I'd like to briefly survey typical examples of nanobiotechnologies which are introduced in sample treatment. Also I specifically demonstrate two different simple techniques to concentrate and detect bacteria from the samples using multifunctional silica nanotube (SNT).

• BMB Reports
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• v.37 no.4
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• pp.387-393
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• 2004

The L-asparaginase (E. C. 3. 5. 1. 1) enzyme was purified to homogeneity from Pseudomonas aeruginosa 50071 cells that were grown on solid-state fermentation. Different purification steps (including ammonium sulfate fractionation followed by separation on Sephadex G-100 gel filtration and CM-Sephadex C50) were applied to the crude culture filtrate to obtain a pure enzyme preparation. The enzyme was purified 106-fold and showed a final specific activity of 1900 IU/mg with a 43% yield. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of the purified enzyme revealed it was one peptide chain with $M_r$ of 160 kDa. A Lineweaver-Burk analysis showed a $K_m$ value of 0.147 mM and $V_{max}$ of 35.7 IU. The enzyme showed maximum activity at pH 9 when incubated at $37^{\circ}C$ for 30 min. The amino acid composition of the purified enzyme was also determined.

• Journal of fish pathology
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• v.17 no.1
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• pp.11-20
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• 2004

The present study compared purification methods of hen egg yolk immunoglobulin (IgY) from the hen immunized with Edwardsiella tarda. The purification of anti-E. tarda IgY was performed by four different methods, polyethylene glycol (PEG), chloroform polyethylene glycol (Chloroform-PEG), ammonium sulfate and purification kit. Purified IgY had heavy chain of 64 kDa and light chain of 27 kDa size. IgY purified from the hen immunized with E. tarda showed higher ELISA values and agglutination titers than those with IgY purified from the non-immunized hen as a negative control. In addition, purified IgY recognized similar E. tarda proteins to those with anti-E. tarda rabbit serum by western blotting. Purified IgY had an agglutination titer of 1:512 by PEG method and ammonium sulfate method, and 1:128 by chloroform-PEG method and purification kit. Moreover, PEG method was the most rapid method among the four different IgY purification methods. These results indicate that PEG method is effective purification method maintaining biological activity of the IgY.