• Journal of Microbiology and Biotechnology
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• v.1 no.4
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• pp.232-239
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• 1991

Caboxymethyl cellulase (CMCase) I was purified from the induced culture filtrate of Penicllium verruculosum F-3 by ammonium sulfate precipitation, DEAE-Sephadex A-50 chromatography and Bio-gel P-150 filtration. The purified enzyme was assumed to be a glycoprotein consisting of 8.5% carbohydrate and having a molecular weight of 70.000 in SDS-polycrylamide gel electrophoresis (SDS-PAGE). The purified enzyme-specific anti-CMCase I IgG was obtained by rabbit immunization and protein A-sepharose CL-4B chromatography. The fungal poly($A^+$) RNA was isolated from the total RNA of the mycelium grown under cellulase induction conditions by oligo(dT)-cellulosse chromatography. The translation products in vitro were prepared by translating the isolated poly ($A^+$) RNA in rabbit reticulocyte lysate and analyzed by SDS-PAGE and fluorography. Of the translation products, CMCase I was identified by the immunoprecipitation against anti-CMCase I IgG.

• Microbiology and Biotechnology Letters
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• v.19 no.3
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• pp.303-307
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• 1991

The hemolyic polypeptide in delta-endotoxin from Bacillus thun'ngiensis subsp. darmstadiensis 73ElO-2 was purified by Sephadex G-IOO gel filtration and DEAE-cellulose ion exchange column chromatography. The purity of hemolysin was confirmed by ouchterlony test and SDS-PAGE. The molecular weight of the purified hemolysin was approximately 64 KDa by SDS-PAGE. The purified hemolysin has not mosquitocidal activity against larvae of Aedes agypti, but hemolytic activity on red blood cells of rat. There is no serological relationship between delta-endotoxin from B. thuringiensis subsp. israelensis and the purified hemolysin from the . strain 73ElO-2.

• YAKHAK HOEJI
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• v.30 no.6
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• pp.343-347
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• 1986

Korean ginseng was purified to obtain radioprotective protein fractions by buffer extraction, ammonium sulfate fractionation, CM-cellulose column chromatography, heat inactivation and Sephadex G-75 column chromatography. The final three fractions, GI, GII and GIII were subjected to Disc-polyacrylamide gel electrophoresis (PAGE) and SDS-PAGE. The molecular weights(M.W.) of native and denatured proteins were estimated by using regression line equations obtained from the mobilities of standard proteins. As the results, in Disc-PAGE, the GI fraction showed two protein bands with M.W. of above 213, 000 and 55, 000, GII showed one band with M.W. of 44, 000 and GIII, also one band with M.W. of 19, 000. In SDS-PAGE, GI fraction gave four subunit bands with M.W. of above 114, 000, 27, 000, 24, 000 and 19, 000, GII gave two bands with M.W. of 46, 000 and 22, 000, and GIII, one band of 19, 000.

• Microbiology and Biotechnology Letters
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• v.13 no.1
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• pp.51-57
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• 1985

Delta-endotoxin in Bacillus thuringiensis var. finitimus, HD-1, HD-9 and HD-73 strains were isolated by NaBr, CsCl and Renografin density gradients. The purity of the toxin was about 98%. The purified o-endotoxin was analyzed by SDS-PAGE, electron microscopic observation and bioassay. According to SDS-PAGE, the molecular weight of subunits of the o-endotoxin were about 66,000 and 130,000 daltons. The shapes of the crystal toxin observed by TEM except finitimus strain were bipyramidal. When the purified endotoxin was bioassayed against tobacco horn worm, the entomocidal activities ($1{\mu}g/ml$) of HD-1 and HD-73 strains were, respectively, 60% and 100% at nine days after treated. The molecular weights of the plasmids isolated from B. thuringiensis were various from 0.5 to 120 Kb. The numbers of plasmids in HD-1, HD-9 and HD-73 strains were 12, 3 and 11, respectively, but B. thuringiensis var. finitimus strain had no plasmid.

• Korean Journal of Food Science and Technology
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• v.37 no.5
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• pp.812-815
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• 2005

Effects of poly-${\gamma}$-glutamate (PGA) on antioxidative characteristics of amino-carbonyl reaction products of glucose were investigated. Rapid browning reaction was observed under heat and alkaline condition (pH 8.2). Browning products were separated by Sephadex G-50, and brown color intensity and electron-donating ability of DPPH (2,2-diphenyl-1-picrylhydrazyl) of each fraction were measured. Fraction-7 (F-7) and -20 (F-20) showed high DPPH scavenging values. UV-VIS absorption spectrum of F-20 was similar to melanoidin peak, and F-7 showed maximum absorption peak at 270 nm. Molecular weight of F-7 was over 35 kDa as determined by SDS-PAGE. Although F-20 could not be measured on SDS-PAGE, its size was smaller than F-7.

• Parasites, Hosts and Diseases
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• v.29 no.1
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• pp.1-8
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• 1991

Out of many component proteins in crude saline extract of Spirometra mansoni plerocercoid (sparganum) , 36 kDa and 29 kDa proteins were found to be the most antigenic and were already purified by immunoaffinity chromatography using monoclonal antibody as a ligand. In this study, a single step purification of these potent antigenic proteins of sparganum extract was investigated. When the crude saline extract was charged to gelatin-Sepharose 4B affinity column, 36 kDa and 29 kDa protein fractions were bound. SDS-polyacrylamide gel electrophoresis (PAGE) and SDS-PAGE/immunoblot confirmed that the bound protein to gelatin was serologically pure. When evaluated by ELISA with patients sera, the purified protein of 36 and 29 kDa also showed improved antigenicity.

• KSBB Journal
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• v.17 no.1
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• pp.33-37
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• 2002

To improve in vitro release kinetic of G-CSF in PLGA microsphere, G-CSF was PEGylated with methoxy polyethylene glycol-aldehyde (mPEG-aldehyde, MW 5000). The majority of G-CSF was mono-PEGylated and it was characterized using SDS-PAGE, HPLC, and peptide mapping. The PLGA microencapsulation with the native, or PEGylated G-CSF was performed using W/O/w method, where the encapsulation efficiency was high. For the high loading of G-CSF to microsphere, G-CSF and PEGylated G-CSF were concentrated and then verified the protein stability using native gel and gel filtration chromatography. In comparison with native G-CSF, PEGylated G-CSF was released during the extended period and its maximum amount of released G-CSF was also increased.

• Reproductive and Developmental Biology
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• v.36 no.3
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• pp.193-198
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• 2012

This study was to examine expression of the recombinant full-length adiponectin (recombinant adiponectin) in insect ovarian cell culture system and to characterize structural properties of the recombinant adiponectin secreted in medium. Gene construct encoding the recombinant adiponectin contained N-terminal collagen-like domain (110 Amino Acids, AAs), C-terminal globular domain (137 AAs) and C-terminal peptides for detection with V5 antibody (26 AAs included adaptor peptide) and purification using the 6xHis tag (6 AAs). The approximate molecular weight of the product (monomer) was 35 kDa. Molecular mass species of the expressed recombinant adiponectin were monomer (~35 kDa), dimer (~70 kDa), trimer (~105 kDa) and hexamer (~210 kDa). The major secreted species were the LMW forms, such as monomer, dimer, and trimer. There was MMW of hexamer as minor form. HMW multimers (~300 kDa) were shown as a tracer or not detected on the SDS-PAGE in several experiments (data not shown). The multimer forms in this study were not compatible to those in animal or human serum and adipose tissue by other researcher's study in which the major multimer forms were HMW. By protein denaturing experiments with reducing reagent (${\beta}$-MeOH), anionic detergent (SDS) and heat ($95^{\circ}C$) on the SDS-PAGE, not all adiponectin multimers seemed to have disulfide bond linked structure to form multimers. The recombinant adiponectin which expressed in insect ovarian cell culture system seemed to have the limitation as full physiological regulator for the application to animal and human study.

• Journal of the Korean Society of Food Science and Nutrition
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• v.19 no.4
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• pp.315-320
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• 1990

Studies on protein and antinutritional factors of rapeseeds are ncessary for effective utiliza-tion of defatted rapeseed meal. Proteins were extracted from seeds of several species of rape-seeds and analyzed by SDS-PAGE and the contents of glucosinolate and phytate were determi-ned. One percent solution extraction and extraction yield was relatively higherfor B. campestris and B. juncea than for other species. SDS-PAGE revealed that rapeseeds of most species were rich in low molecular weight proteins and that in particular the roteins of B. napus var. Halla and B. juncea were composed of simpler subunits as compared with other species. The content of glucosinolate was around 10mg/g of defatted meal for B. juncea however for var. Halla it was 7.26mg/g of defatted meal and 0.46mg/g of protein concentrate which were the lowest values. The level of phytate was between 2.7 and 4.6% for all species tested. Our results indicate thT B, napus var. Halla is the desired species for the utilization of rapeseed proteins.

• Parasites, Hosts and Diseases
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• v.34 no.1
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• pp.35-48
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• 1996

Non-biting midges fchironomidae, Dipteral are one of the largest insect families, which are distributed worldwidely and are found in nearly all types of inland waters. They are known to be aggressive inhalant allergens which cause allergenic diseases. In this study, the crude antigens of Chironomus SavipLumn adults which are most widely distributed in Korea were extracted. and their allergens were analysed with the sera from experimentally sensitized mice. The mice were immunized with $1{\;}\mu\textrm{g}{\;}or{\;}10{\;}\mu\textrm{g}$ of the crude antigens, respectively, and the specific serum IgE levels were measured by both ELISA and passive cutaneous anaphylaxis (PCA) techniques. The highest levels of both total IgE and chironomid-specific IgE were found in the mouse sera obtained after 9 weeks of the first infection with $1{\;}\mu\textrm{g}$ crude antigen. The crude antigen was separated into 16-18 protein bands on gel by SDS-PAGE. The crude extract was assessed by SDS-PAGE/immunoblot analysis. One IgE-binding band (65 kDa) was detected by developing with colorimetric substrate, and 4 IgE-binding bands (65, 52, 35 and 25 kDa) by developing with CSPD chemiluminescent substrate. The SDS-PAGE gel of the crude extract of chironomid adults was equally cut into 30 pieces and each of them was eluted to isolate proteins by molecular weight, and the allergenicity of each eluate was assessed by applying P-K test on rats. Proteins of 65, 35 and 15 KDa showed the highest P-K titer (${\times}512$) which was 16 times higher than that of the crude extract (${\times32}$). The P-K titer of 52 kDa protein was also 4 times higher ($128{\times}$) than that of the crude extract, whereas the 25 kDa protein poorly responded, which seemed not antigenic. In conclusion, the present result in mice demonstrated that adults of Chironomus fcuiplumus, a predominent species in Korea, cause allergenic diseases and the main allergens are 65, 52, 35 and 15 kDa proteins, of which 65 kDa protein seems to be a main allergen.