• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.7
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• pp.1186-1191
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• 2004

Bacillus sp. $\beta$-mannanase was purified by DEAE-sephadex ion exchange column chromatography. The partially purified P-mannanase exhibited maximum activity at pH 6.0 and 5$0^{\circ}C$, and was stable at a pH range of 5.5 to 7.0, and at temperature between 30 to 5$0^{\circ}C$. Konjac glucomannan was hydrolyzed by the purified $\beta$-mannanase, and then hydrolysates separated by 1st activated carbon column chromatography and 2nd sephadex G-25 gel filtration. The main hydrolysates were composed of D.P 5 and 7 glucomannooligosaccharides by TLC and FACE method. To investigate the effects of guar gum glucomannooligosaccharides on the in vitro growth of B. longum, B. bifidum, B. infantis, B. adolescentis, B. animalis, and B. breve, Bifidobacterium spp. were cultivated individually on the modified-MRS medium containing carbon SOUTce such as D.P 5, and D.P 7 glucomannooligosaccharides, respectively. B. longum grew up 4.6-fold and 5.3-fold more effectively by the replacement of D.P 5 and 7 glucomannooligosaccharides as the carbon source in a comparasion of standard MRS. Also, B. breve and B. animalis slightly grew up by the treatment of D.P 5 glucomannooligosaccharide.

• Microbiology and Biotechnology Letters
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• v.32 no.2
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• pp.117-122
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• 2004

Bacillus sp. $\beta$-mannanase was purified by DEAE-sephadex ion exchange column chromatography. The specific activity of the purified enzyme was 21.57 units/$m\ell$ protein, representing an 95.33-folds purification of the original crude extract. The final preparation thus obtained showed a single band on SDS-polyacrylamide gel electrophoresis. The molecular weight was determined to be 38.9 kDa. Guar gum galactomannan was hydrolyzed by the purified $\beta$-mannanase, and then the hydrolysates was separated by activated carbon column chromatography and Sephadex G-25 gel filtration. The main hydrolysates were composed of D.P. (Degree of Polymerization) 5 and 7 galactomannooligosaccharides. To investigate the effects of guar gum galactomannooligosaccharides on in vitro growth of Bifidobacterium longum, B. bifidum, B. infantis, B. adolescentis, B. animalis, and B. breve, Bifidobacterium spp. were cultivated individually on the modified-MRS medium containing carbon source such as D.P. 5 and D.P. 7 galactomannooligosaccharides, respectively B. longum and B. bifidum grew up l0-fold and 9.8-fold more effectively by the treatment of D.P. 5 galactomannooligosaccharides, compared to those of standard MRS medium. Especially, D.P. 5 was more effective than D.P. 7 galactomannooligosaccharide on the growth of Bifidobacterium spp.

• Food Science and Preservation
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• v.10 no.2
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• pp.246-251
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• 2003

Pretense produced by Bacillus subtilis PANH765 was purified from culture supernatant by using ammonium sulfate fractionation DEAE-cellulose ion exchange chromatography, and gel filtration with Sephacryl S 200 HR and Sepharose CL-6B. DEAE-cellulose ion exchange column chromatography, separated the pretense into one fraction. This fraction was further purified using Sephacryl S 200 HR and Sepharose CL-6B gel titration. The molecular mass of pretense was estimated to be 35.0 kDa by the SDS-PAGE and gel filtration using Sepharose CL-6B. The results indicated that the purified pretense are monomeric proteins. Specific activity and purification folds of pretense were 657 U/mg and 4.35, respectively. The optimum temperature, optimum pit stable at a temperature range and pH ranges for the purified protease were 65$^{\circ}C$, 7.05, 50 ∼ 75$^{\circ}C$ and 6.0 ∼ 7.5, respectively. The pretense activity was decreased by the presence of PMSF and DFP, which the protease activity was increased by the presence of Na$\^$+/, K$\^$+/, Mg$\^$2＋/ and NH$_4$$\^$+/ ions.

• Journal of Dairy Science and Biotechnology
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• v.15 no.1
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• pp.1-9
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• 1997

This study was conducted to efficiently separate the Ig from Korean native cattle colostrum and to utilize them as an immunogen for the production of antibodies aginst rabbit. The results obtained were as follows : 1. About 84% of Ig G could be separated from Korean native cattle colostrum by·gel filtration using Superose 12 column on HPLC. The separation profile of Korean native cattle colostral immunoglobulin was similar that of Holstein colostral Ig. 2. Separation of Korean native cattle colostral Ig by anion exchange chromatography using Mono Q column on HPLC was poor resolution chromatographic pattern. 3. Hi-Trap Protein G column showed better results than the Protein A Sepharose CL-4B column in the Ig G binding capacity from Korean native cattle colostral Ig. 4. Protein G Sepharose Fast Flow system resulted in higher Ig g binding capacity as the industrial size scale-up approach. 5. Sufficient titer reaction of antibody to Korean native cattle colostral Ig G was confirmed by ELISA.

• Analytical Science and Technology
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• v.11 no.4
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• pp.231-234
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• 1998

Separation factor for $^6Li$ and $^7Li$ has been determined using ion exchange resin having 1,7,13-trioxa-4,10,16-triazacyclooctadecane ($N_3O_3$) as an anchor group. The ion exchange capacity of the $N_3O_3$ ion exchanger was 2.0 meq/g dry resin. The lighter isotope, $^6Li$, is concentrated in the fluid phase, while the heavier isotope, $^7Li$, is enriched in the resin phase. By column chromatography [0.3 cm(I.D)${\times}$30 cm (height)] using 3.0 M ammonium chloride solution as an eluent, single separation factor, ${\alpha}$, 1.018, i.e. $(^7Li/^6Li)_{resin}/(^7Li/^6Li)_{fluid}$ was obtained by the Glueckauf theory from the elution curve and isotope ratios.

• Korean Chemical Engineering Research
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• v.50 no.4
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• pp.713-717
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• 2012

Approximately forty proteins in egg white have been widely studied for their functional properties. To develop a procedure of separation for pure and non-altered proteins from egg white, purification study was conducted to isolate lysozyme, ovotransferrin, and ovalbumin. Ion exchange cartridge can selectively separate proteins from egg white, and reversed-phase HPLC (RP-HPLC) could identify separated proteins. Proteins in egg white were purified by HI trap ion exchange cartridge SP and Q with buffers pH 8.0 and 5.2. C18 column (Phenomenex, USA) was used for RP-HPLC analysis and isocratic mobile phase was used with acetonitrile (ACN)/distilled water (DW)/trifluoroacetic acid (TFA) in the ratio of 50/50/0.1. Comparing the retention times of standards in RP-HPLC experiments showed that ovotransferrin, ovalbumin, and lysozyme in egg white were eluted successively in the RP-HPLC column after the pretreatment in SP and Q ion exchange cartridges.

• The Korean Journal of Food And Nutrition
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• v.11 no.6
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• pp.629-634
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• 1998

1-Deoxynojirimycin which is a potent intestinal ${\alpha}$-glucosidase inhibitor was purified from the culture broth by ion exchange chromatography, Sephadex LH20 column chromatography, TSK gel chromatography and HPLC respectively. Acute toxicity of 1-deoxynojirimycin, which was loaded through the oral as dose of 200mg/kg, was investigated in IRC mouse. None of the tested IRC mice were not dead and increase of body weight showed also the same results in comparison with control mice. The antimicrobial susceptibility of 20 pathogenic strains against 3 antidiabetic compounds (1-deoxynojirimycin, AO-128, acarbose) were obtained by agar dilution method. All of the three antidiabetic compounds has very weak antimicrobial activity (MIC>100$\mu\textrm{g}$/ml).

• 한국생물공학회:학술대회논문집
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• 2002.04a
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• pp.510-513
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• 2002

Iron is an essential nutrient for most organisms, which supplied to them in a protein-iron complex known as ferritin. Ferritins are multimeric proteins those are consisted of spherical shell of 24 subunits defining a cavity of about 8nm in diameter. Soluble form of ferritin was separated from disrupted cells, followed by silica powder adsorption. Ferritin was recovered from silica-poweder by distiiled water, which was applied to DEAE anion exchage chromatography. Collected fractions from the DEAE column were assayed to gain the amount and the purity of ferritin by using GF-HPLC.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1995.04a
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• pp.126-126
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• 1995

본 연구는 중금속에 의한 생체내 독성기전을 연구하고자 흰쥐간의 상층액에서 카드뮴과 잘 결합하는 HMWP들을 분리, 정재하고 나아가 생화학적 특성을 밝히고 이단백질이 카드뮴에 의해 생성되는 것인지 아니면 간의 기존 단백질인지를 밝히고자 함을 연구의 목적으로 하였다. CdCl %2 (3mg/kg body wt.)을 3일간 ip injection시킨 후 흰쥐의 간을 적출하고 균질화하여 원심분리한 crude extract를 직접 Sephacryl S-100에 흡착시켜 10mM phosphate buffer(pH 7.0)으로 용출시켰다. 용출된 fraction tube를 UV spectrometer로 흡광도를 측정하고 atomic absorption spectrophotometer로 카드뮴량을 측정하여 카드뮴량이 높은 분획을 모아서 ion exchange column chromatography(DEAE-Sepharose)에 흡착시켜 염농도 구배로 chromatography를 실시하고, 음이온 교환수지에서 흡착되지 않고 용출된 분획을 ultrafiltration으로 농축시킨 후 S-Sepharose에 흡착시켜 염농도 구배로 chromatography를 실시한 결과 두 종류의 카드뮴 결합 단백질(Cd-BP)를 분리, 정제하였다.

• YAKHAK HOEJI
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• v.52 no.1
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• pp.79-84
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• 2008

Alismatis Rhizoma has been used as diuretics and antiphlogistics in the Chinese oriental medicine. A trypsin inhibitor was isolated from Alismatis Rhizoma using DEAE ion exchange column, trypsin affinity column, and FPLC chromatography, and its activity and characteristics were studied. The purifed Alismatis Rhizoma trypsin inhibitor (ARTI) was estimated to be about 22 kDa. The sequence determination on N-terminal amino acid residues and 84 amino acid residues has been completed, yet no homology has been found with trypsin inhibitors reported at NCBI. ARTI did not show inhibitory activities on chymotrypsin and elastase, however it exhibited a significant inhibitory activity on bovine trypsin, and formed a complex with rat liver 10-formyltetrahydrofolate dehydrogenase.