• Journal of Life Science
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• v.6 no.4
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• pp.250-263
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• 1996

In this study Carotenoprotein from Styela clava were extracted, and purified by ammonium sulfate fraction, Sephadex G- 200 gel chromatography and DEAE-cellulose ion exchange chromarography. The purified carotenoprotein from styela clava had absorption maxima of 487nm, 463nm and 280nm, and the carotenoid liberated from carotenoprotein had 478nm and 452nm with inflexion. One miligram of caroteno-protein contained 0.35 $\mu$g of carotenoid. The carotenoprotein had an approximate molecular weight of 398 kDa (gel filtration). SDS-PAGE showed only a single polypeptide chain with a molecular weight of 62.4 kDa. The amino acid composition of the carotenoprotein were mainly glutamic acid(11.48%), valine(10.75%), leucine (10.45%), aspartic acid(9.94%), while cysteine and tryptophan were not found. The carotenoprotein contained lipid as structure units. In the carotenoprotein, the major fatty acids were oleic acid, plamitoleic acid and palmiunsaturated fatty acids 33.6%, saturated fatty acids 18.6%. In addition, the levels of higher unsaturated fatty acids were high as much as 30.8% of the total fatty acids. Carotenoid was extracted from the carotenoprotein by acetone. Thin layer chromatography showed only carotenoid to be present. Its chemical reactivities and spectroscopic properties were studied and elucidated as astaxanthin.

• Journal of Embryo Transfer
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• v.13 no.1
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• pp.43-52
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• 1998

The embryogenesis stimulating activity(ESA) had been shown in co-culture of embryos with bovine oviduct epithelial cell(BOEC) and culture in BOEG-conditioned medium. The present study was undertaken to purify and quantify the embryotropic proteins and to determine the optimum concentration of the embryotropic protein for the proper development of embryos. In BOEC-conditioned medium, five major bands of proteins were detected(66, 53, 40, 32 and 24 kDa) by SDS-PAGE. From these proteins, 288pg of protein that had a 32kDa molecular weight was purified by gel filtration column and perfusion chromatography ion-exchange column. When purified protein was supplemented to the in vitro culture media at various concentrations in protein-free media, 2.5$\mu$g /ml supplement group showed significantly higher rates of embryo development into morula /blastocyst stages than other groups(p<0.05). In conclusion, we purified 32kDa protein from BOEC-conditioned medium and this protein showed optimum embryogenesis stimulating effect at 2.5$\mu$g /ml.

• Journal of Life Science
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• v.17 no.2 s.82
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• pp.254-259
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• 2007

Biochemical characterization of lectin isolated from fruit of cherry tomato through neutral saline extraction, ammonium sulfate precipitation, and affinity chromatography on Sephadex G-200 was studied. The lectin was agglutinated by trypsin-treated human ABO erythrocytes, and the most pronounced activity of agglutination was observed at B type erythrocyte. The analysis of the lectin by SDS-PAGE showed the high intensity band with molecular weights of 10.7 kDa. The optimal temperature and thermal stability of the lectin was $40^{\circ}C$ and $40-60^{\circ}C$, respectively. The maximal pH of this lectin was pH 7.2.

• Journal of Integrative Natural Science
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• v.10 no.3
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• pp.154-161
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• 2017

An antimicrobial peptides-producing Bacillus strain CBS73 was isolated from fermented food (kimchi) that produces low-molecular-weight proteins with broad-spectrum antimicrobial activity. Our goal was to explore the therapeutic potential of antimicrobial substances produced by Bacillus species. Peptide CBS73 was purified from Bacillus subtilis subsp. subtilis with identity of 99.79%. It was found to be stable at pH 4.0-10.0 and temp $20-60^{\circ}C$. A protein band around 5.2 kDa was detected in tricine-SDS-PAGE and band was confirmed by MALDI-TOF test. Peptide CBS73 showed antimicrobial activity against MDR bacteria. The minimal inhibitory concentration (MIC) of peptide CBS73 for vancomycin-resistant S. aureus (VRSA), vancomycin resistant Enterococci (VRE) and Salmonella typhimurium ranged from $10-40{\mu}g/mL$. The antioxidant activity of peptide CBS73 was measured by DPPH scavenging, reducing power activity and total phenolic content. Cell viability and NO production result showed less cytotoxic effect upto $12{\mu}g/mL$. Peptide CBS73 could be a promising antimicrobial agent for clinical application.

• Journal of Pharmacopuncture
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• v.9 no.2
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• pp.105-111
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• 2006

Objectives : This study was conducted to carry out Purification of Melittin and other peptide components from Bee Venom using gel filtration chromatography and propionic acid/urea polyacrylamide gel electrophoresis Methods : Melittin and other peptide components were separated from bee venom by using gel filtration chromatography on Sephadex G-50 column in 0.05M ammonium acetate buffer. Results : Melittin and other peptide components were separated from bee venom by using gel filtration chromatography on Sephadex G-50 column in 0.05M ammonium acetate buffer. The fractions obtained from gel filtration chromatography was analyzed by using SDS-PAGE and propionic acid/urea polyacrylamide gel electrophoresis. The melittin obtained from the gel filtration contained residual amount of phospholipase $A_2$ and a protein with molecular weight of 6,000. The contaminating proteins were removed by the second gel filtration chromatography. Conclusion : Gel filtration chromatography and propionic acid/urea polyacrylamide gel electrophoresis are useful to separate peptide components including melittin from bee venom.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.10 no.3
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• pp.620-624
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• 2009

Apolipophorin-III (apoLp-III) was isolated and purified from the last instar larval hemolymph of Galleria mellonella by gel chromatography (Sephadex G-100) and ion exchange chromatography (CM-52). In the present study, I wanted to show that apoLp-III is taken up into the adult ovary in Galleria mellonella. Adult ovary tissues were incubated at room temperature for 30 min with fluorescein isothiocyanate (FITC)-labeled apoLp-III. Fluorescence microscopy and sodium dodecylsulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) revealed that adult ovary tissues internalize fluorescence-labeled apoLp-III. The results suggest that apoLp-III is taken up by the adult ovary.

• Macromolecular Research
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• v.15 no.3
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• pp.205-210
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• 2007

Collagen is the major structural protein of connective tissues. It can be used as a prosthetic biomaterial applicable to artificial skin, tendon, ligaments, and collagen implants. The objective of this study is to investigate the possibility of realizing wound dressing medical products by the synthesis of composite materials with collagen and a biodegradable polymer, PLLA, via a surface modification process. Type I collagen was obtained from pig skin by a separation process. The structural characteristics of the extracted collagen were confirmed by SDS-polyacrylamide (PAcr) gel electrophoresis (PAGE) and FTIR. Also, PLLA-g-PAcr was synthesized by the radical polymerization of acrylamide initiated by AIBN in the presence of PLLA. The surface of PLLA was modified by the presence of the acrylamide residues. The structural characteristics of the copolymer were analyzed by FTIR, $^1H-NMR$ and contact angle measurements. The water uptake and WVTR of the collagen/PLLA-g-PAcr composite tended to increase with increasing collagen concentration and with decreasing EDC concentration.

• BMB Reports
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• v.33 no.6
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• pp.483-492
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• 2000

The Herpes simplex virus type 1 (HSV-1) strain F glycoprotein H (gH) gene in the pHLB-4 plasmid was recombinated into a baculovirus expression vector (lacZ-HcNPV) to construct a recombinant virus GH-HcNPV expressing gH. The sequences of gH and its expression were analyzed. The gH gene was located in the 6.41 kb BglII fragment. The open reading frame (ORF) of the gH gene was 2,517 bp and codes 838 amino acid residues. Insect cells infected with this recombinant virus synthesized a high level of the matured and gX-gH fusion protein with approximately 112 kDa. The fusion gH protein was localized on the membrane of the insect cells as seen by using immunofluorescence assay and accumulated in the cultured media by the SDS-PAGE and immunoprecipitation assays. The amino acid sequence presents additional characteristics compatible with the structure of a viral glycoprotein: signal peptide, putative glycosylation sites and a long C-terminal transmembrane sequence. Antibodies raised in mice to this recombinant protein recognized viral gH and neutralized the infectivity of HSV-1 in vitro. These results demonstrate that it is possible to produce a mature protein by gene transfer in eukaryotic cells, and indicate the utility of the HcNPV-insect cell system for producing and characterizing eukaryotic proteins. Furthermore, the neutralizing antibodies would appear to protect mice against HSV; accordingly, this particular recombinant protein may be useful in the development of a subunit vaccine.

• Journal of Microbiology and Biotechnology
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• v.19 no.2
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• pp.194-203
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• 2009

$HpaG_{Xooc}$, from rice pathogenic bacterium Xanthomonas oryzae pv. oryzicola, is a member of the harpin group of proteins, eliciting hypersensitive cell death in non-host plants, inducing disease and insect resistance in plants, and enhancing plant growth. To express and secret the $HpaG_{Xooc}$ protein in Bacillus subtilis, we constructed a recombinant expression vector pM43HF with stronger promoter P43 and signal peptide element nprB. The SDS-PAGE and Western blot analysis demonstrated the expression of the protein $HpaG_{Xooc}$ in B. subtilis. The ELISA analysis determined the optimum condition for $HpaG_{Xooc}$ expression in B. subtilis WBHF. The biological function analysis indicated that the protein $HpaG_{Xooc}$ from B. subtilis WBHF elicits hypersensitive response(HR) and enhances the growth of tobacco. The results of RT-PCR analysis revealed that $HpaG_{Xooc}$ induces expression of the pathogenesis-related genes PR-1a and PR-1b in plant defense response.

• Applied Biological Chemistry
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• v.51 no.3
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• pp.159-163
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• 2008

Xylogone sphaerospora ${\beta}$-mannanase was purified by Sephadex G-100 column chromatography. The specific activity of the purified enzyme was 8.44 units/ml protein, representing an 56.27-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 42kDa. Konjac glucomannan was hydrolyzed by the purified ${\beta}$-mannanase, and then the hydrolysates was separated by activated carbon column chromatography. The main hydrolysates were composed of D.P. (Degree of Polymerization) 3 and 4 glucomannooligosaccharides. For elucidate the structure of D.P 3 and 4 glucomannooligosaccharides, sequential enzymatic action was performed. D.P 3 and 4 were identified as M-G-M and M-M-G-M (G- and M- represent glucosidic and mannosidic link-ages). To investigate the effects of konjac glucomannooligosaccharides on in vitro growth of Bifido-bacterium longum, B. bifidum, B. infantis, B. adolescentis, B. animalis, B. auglutum and B. breve. Bifidobacterium spp. were cultivated individually on the modified-MRS medium containing carbon source such as D.P. 3 and D.P. 4 glucomannooligosaccharides, respectively. B. longum and B. bifidum grew up 3.9-fold and 2.8-fold more effectively by the treatment of D.P. 4 glucomannooligosaccharides, compared to those of standard MRS medium. Especially, D.P. 4 was more effective than D.P. 3 glucomannooligosaccharide on the growth of Bifidobacterium spp.