• Journal of Sericultural and Entomological Science
• /
• v.45 no.2
• /
• pp.90-95
• /
• 2003

This study was carried out to identify of C3G (cyanidin-3-glucoside) from mulberry fruits and quantify with different varieties. C3G of mulberry fruits was extracted with 1％ HCl-MeOH and purified with open column (5${\times}$90cm) which filled with Amberlite IRC-50 ion exchange resin. The $\lambda$max ranges of the purified C3G on UV/vis spectrum were 516nm and 280nm. Also, molecular weight of C3G from mulberry fruits by LC-Mass was determined as 449. From above results, we concluded that anthocyanin pigment of mulberry fruits was C3G only. The cyanidin-3-glucoside (C3G) was separated and quantified by High Performance Liquid Chromatography (HPLC) system using a Nova-Pack C$\_$18/ column. Mean content of the 35 tested accessions was 0.89％. Also fruity characteristics as well as C3G content to select the desirable mulberry varieties for the production of fruit were researched and analyzed. We selected three suitable varieties such as 'Susungppong', 'Kangsun', and 'Jeolgokchosaeng(Chungpuk)'.

• Journal of Microbiology and Biotechnology
• /
• v.9 no.4
• /
• pp.469-474
• /
• 1999

Thermoactinomyces sp. E79 produced two types of thermostable alkaline proteases extracellularly. A minor protease was separated from a major protease by using DEAE-column chromatography. This enzyme was purified to homogeneity by ammonium sulfate and DEAE-Sepharose ion-exchange chromatography. The purified minor protease showed different biochemical properties compared to the major protease. The molecular mass of the purified enzyme was estimated by SDS-PAGE to be 36 kDa. Its optimum temperature and pH for proteolytic activity against Hammarsten casein were $70^{\circ}C$ and 9.0, respectively. The enzyme was stable up to$75^{\circ}C$ and in an alkaline pH range of 9.0-11.0. The enzyme was inhibited by phenylmethylsulfonyl fluoride (PMSF) and $Hg^{2+}, indicating that the enzyme may be a cysteine-dependent serine protease. In addition, the enzyme cleaved the endoproteinase substrate, succinyl-Ala-Ala-Pro-Phe-p- nitroanilide, and the $K_m$ value for the substrate was 1.2 mM.

• The Korean Journal of Food And Nutrition
• /
• v.10 no.4
• /
• pp.503-508
• /
• 1997

The extraction and separation methods of protein-bound polysaccharides from the mycelium and culture broth of L. edodes were investigated. The use 2% solution of surface active agent, Triton X-100 was effective for extraction of the protein-bound polysaccharide from the mycelium. The extraction of the protein-bound polysaccharides from mycelium with hot water was achieved by 4 hours extraction at 10$0^{\circ}C$. For the separation and partial purification of the protein bound polysaccharides the column chromatography using DEAE-Cellulose, DEAE-Sephadex and Sephadex proved to be effective.

• Journal of Plant Biology
• /
• v.35 no.2
• /
• pp.99-106
• /
• 1992

The endosperm protein, vicilin, of ginseng (Panax ginseng C.A. Meyer) was purified by ammonium sulfate precipitaion, gel permeation and ion exchange column chromatography. Vicilin is a glycoprotein composed of 2 subunits with molecular masses of 55,000 (large subunit) and 44,000 (small subunit). The anti-vicilin antibody was raised in rabbit, and purified by DEAE Affi-Gel Blue affinity chromatography. The endosperm cells of the seed were reacted with this anti-vicilin antibody and colloidal gold conjugated secondary antibody. Gold particles were labelled on the elaborating granules of Golgi complex, electron-dense granules and protein bodies in the endosperm cells. These results indicated that the vicilin, which was synthesized in rough endoplasmic reticulum and transported to Golgi, was elaborated in saccules of the Golgi and then transported into protein bodies by electron-dense granules.anules.

• Journal of Microbiology and Biotechnology
• /
• v.5 no.4
• /
• pp.206-212
• /
• 1995

A strain BL-29, which produces a extracellular lytic enzyme on E. coli was isolated from the soil. The strain was identified as belonging to the genus Bacillus sp. The lytic enzyme was purified to homogeneity by ion exchange chromatography and gel filtration. Specific activity of the purified enzyme was 28, 850 U/mg protein and yield of the enzyme was 5$%$. The purified enzyme showed a single band on SDS-PAGE and its molecular weight was estimated to be 31, 000 by SDS-polyacrylamide gel electrophoresis and gel filtration column chromatography. The optimum temperature and pH were $55^{\circ}C$ and pH 10.0, respectively. The enzyme was stable at $45^{\circ}C$ but enzyme activity was reduced by up to 50$%$ when the temperature was raised to $55^{\circ}C$ for 15 min. Stable range of pH was from 5.0 to 11.0. but Enzyme activity was inhibited by lead-acetate, mercuric chloride, ethylene glycol-bis-[$\beta$-aminoethyl ether]-N, N, $N^1, $N^1$-tetraacetic acid (EGTA), and ethylenediamine tetraacetic acid (EDTA), but not affected considerably by treatment with other chemical reagents.

• Journal of the Korean Wood Science and Technology
• /
• v.28 no.4
• /
• pp.29-40
• /
• 2000

Highly purified Cerrena unicolor laccase (benzenediol:oxygen oxidoreductase, EC 1.10.3.2) caused the demethoxylation of milled wood lignin and several lignin related substances. The constitutive form of the enzyme produced extracellularly by C. unicolor fermenter culture was isolated and purified by ion-exchange chromatography on the DEAE-Toyopearl column and by affinity chromatography on a ConA-Sepharose and Syringyl-AH-Sepharose 4B columns. The enzyme was further immobilized on functionalized porous glass (CPG) and keratin coated CPG. The demethylating activity was monitored both by estimation of released methanol and by detection of the level of methoxyl groups (also in some water miscible solvents) after incubation of lignin materials with laccase preparations (free and immobilized). The effects of the incubation time and temperature on the demethoxylating activity of immobilized laccase preparations were also studied.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.21 no.6
• /
• pp.725-730
• /
• 1992

The internal invertase was purified from cell free extract of Rhodosporidium toruloides IFO 0559-M-919 by acid precipitation, ion-exchange chromatography and gel filtration to the unique enzyme protein on disc electrophoresis. We have found out that molecular weight of purified internal invertase was 90,000 by gel filtration and the purified enzyme was protein with 4 homogeneous subunits appearing as single band of 22,000daltons on SDS-polyacrylamide gel electrophoresis.

• Korean Journal of Microbiology
• /
• v.26 no.2
• /
• pp.73-81
• /
• 1988

$\alpha$-Amylase (EC.3.2.1.1) of Neurospora crassa (ATCC9279) was cloned in E. coli HB101 using shotgun method, and the enzymes isolated from both N. crassa and E. coli were compared. Chromosomal DNA isolated from the spores of N. crassa was partially digested with PstI restriction endonuclease and rejoined to pBR322 which had been digested with the same enzyme. The resulting recombinant DNA were introduced into E. coli HB101 which had competancy by treating with $CaCl_{2}$. As the result, about 8000 colonies which showed tetracycline resistance were selected and two of the colonies which had 13.5Kb recombinant plasmid exhibit starch degrading activity on starch-containing plate when treated with D-cycloserine. $\alpha$-Amylases from both N.crassa and E. coli were isolated by using ammonium sulfate precipitation, DEAE-cellulose ion exchange column chromatography and Bio-Gel P150 gel foltration column. As the result, about 81.3 fold and 5.6 fold purifications in specific activities were obtained respectively, and specific activities of the gel filtrates were 6.1u/mg and 85u/mg respectively. The properties of both enzymes were compared and they showed quite the similar patterns in optimal temperature, optimal pH and had same molecular weight about 100,000 daltons on gel filtration method. Optimal temperatures for both enzymes were $70^{\circ}C$ and optimal pH were about 6 and 10.

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

• Applied Biological Chemistry
• /
• v.1
• /
• pp.55-61
• /
• 1960

The amino acid compositions of the protein and the nonprotein fractions obtained from a marine brown alga, 'Undaria pinnatifida' were determined by use of Ion Exchange Column Chromatography. The protein nitrogen in the alga was about ten times of the nonprotein nitrogen. Nonprotein fraction obtained from the extraction with 80 percent ethanol contains considerable amount of tree citrulline. Alanine content in the alga was the highest (about 1 per cent in dry weight) and one third of which was found in free state. The amino acid composition of the alga was well balanced and the content of the essential amino acids were relatively higher, than soybean protein. In addition, several peptide like substances were fractionated from nonprotein fraction, in which one way identified as a naturally occurring new tripeptide composed of alanine, glutamic acid and aspartic acid, and the remaining unknown substances are under investigation for the further information.