• Journal of the Korean Chemical Society
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• v.47 no.4
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• pp.363-369
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• 2003

Selenized yeast (Se yeast) containing $0.1{\%}$(w/w) of selenium was obtained when the yeast was incubated at a selenium concentration of 1$1.14{\times}10_-3 M$ in rich medium. After washing several times, the inorganic selenium on the cell wall was confirmed with MBRT. There was no indication of inorganic selenium on the cell wall when the blue color in MBRT was stayed for 15 minutes. The selenized yeast was sonicated, then the selenium contained protein was obtained after salting out by ammonium sulfate at the concentration $80{\%}$ saturation. The seven protein bands were seperated by SDS-PAGE and the selenium concentration in protein was measured by ICP-AES. Analytical data showed that the large expressed protein band contained a relatively large amount of selenium. The proteins of the 47kDa was contained the concentrations of 69.5 ${\mu}$ Se/g of most many content. The protein (47 kDa) was seperated from PVDF membrane by tank-electroblotting. The isolated protein was hydrolyzed under acid condition and reacted with PITC. The derivatives of amino acids were analyzed by HPLC and compared with the data obtained from regular yeast. The resulting selenium-yeast was analyzed with the selenomethionine concentration of $2{\%}$ comparaed with general amino acids. The goal of this study is to analyze the selenium concentration in protein bands and measure the degree of biotransformation of selenomethionine in a specific protein.

• Korean Journal of Animal Reproduction
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• v.16 no.4
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• pp.289-299
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• 1993

The polypeptide patterns of cellular and follicular components were analysed by SDS-PAGE and two dimensional(2-D)electrophoresis combined with isoelectric focusing (IEF) to establish protein profiles in each of the components in porcine follicles. Oocyte-cumulus complexes were cultured in M16+FCS+Gn at 39 in an atmosphere of 5% CO$_2$, in air for 35 h. At the end of the culture, the zona-free oocyte, ZP alone and cumulus cells were prepared and analysed either on 10% SDS-PAGE for the protein profile at the first dimensional gel or 2-D protein pattern. The amounts of each samples were determined for the visualization with Coomasie brilliant blue (CBB) or silver staining, thus giving useful information for the identification of specific proteins in the components or appropriate amount of samples for proper visualization. Oocyte showed 25 and 114 kd major protein band. Other minor components were additionally visualized with CBB on the same gel after silver staining procedure. Cumulus cells also showed specific proteins which is not present in the oocytes. The number of cumulus cell was proper to give major bands with CBB and additional minor bands with silver staining. To establish the degree of contamination from the remnant of the corona radiata to the ZP, zonae were differently prepared or analysed by SDS-PAGE.The preparation of the ZP in this study did not showed any contamination judged by the protein profile of the components. Also follicular fluid showed its specific protein profile without any significant differences among the different sizes of follicles. The established protein profile of each follicular component should be helpful for the identification and elimination of contaminated components, i. e., antigen preparation or immunological studies. The results also suggest that the preparation of each components in the study was appropriate and can be used for a further sensitive biochemical analysis in mammalian oocytes and early embryos.

• Korean Journal of Food Science and Technology
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• v.20 no.6
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• pp.883-889
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• 1988

The salted small shrimps(Acetes japonicus) were fermented for 3 months at room temperature. During the period of fermentation, the changes of shrimp protein properties were determined. The extractability of soluble protein was slightly decreased in 1 month fermentation, but thereafter increased. The contents of 10% TCA soluble fraction were gradually increased during 3 month fermentation, and the rate of 10% TCA soluble fraction/total soluble protein was also greatly increased during the period of fermentation. Sephadex G-100 gel filtration pattern was changed after 1 month fermentation, showing the disappearance of low molecular weight protein peaks, the decomposition and the delay of elution time of main shrimp protein peaks. Polyacrylamide gel disc electrophoresis patterns showed the degradation of main protein bands into lots of smaller bands after 1 month fermentation. The contents of total free amino acids were slightly decreased in 1 month fermentation and then gradually increased during the Period of fermentation. The rate of free amino acids/soluble protein was steadily increased during the period of fermentation, but the rate of free amino acids/10% TCA soluble fraction was decreased continually during the period of fermentation. The contents of most free amino acids were increased during the period of fermentation, but those of histidine and arginine were greatly decreased in 1 month fermentation. Ammonia was increased after 1 month fermentation. The pH value of salted shrimp was slowly changed during 3 months of fermentation, showing increase from 7.8 to 8.2.

• Journal of Life Science
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• v.8 no.3
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• pp.318-325
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• 1998

To investigate changes in protein and total lipid contents, seed storage protein pattern, and fatty acid composition of germination perilla(Perilla frutescens) seeds. Also, the corresponding value components in cotyledons, hypocotyles and roots were measured according to germination stage. The results were summarized as follows ; During germination, pertein and total lipid contents of Yepsilldalggae and Kwangyang cultivar were decreased continuously. In particular, protein contents rapidly decreased to the 3 days after germination(DAG), and then total lipid contents rapidly decreased between 3 DAG and 10 DAG. In changes of protein and total lipid contents of cotyledons, protein contents of Yeupsildalggae was increased during the germination, but Kwangyang cultivar was decreased during the same periods. The total lipids contents of Yeupsildalggae and Kwangyang cultivar were decreased during the germination. According to SDS-PAGE analysis, there was no detectible polypeptide bands on the gel before seed germination suggesting that this may be due to the rapid degradation of the storage proteins in the mature seed by hydrolyttic enzymes during the stage. During germinatation , the polypeptide band with 27$\sim$28KD of Yeupsildalggae and Kwangyang cultivar were accumulated gradually. In changes of fatty acid composition of total lipid of Yeupsildalggae and Kwangyang cultivar , saturated fatty acids such as palmitic acid and stearic acid increased during the germination. On the other hand, unsaturated fatty acid such as linoleic acid and linolenic acid decreased during the same periods. However, oleic acid increased to the 5 DAG, and then was repidly decreased.

• Journal of Plant Biology
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• v.36 no.4
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• pp.377-382
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• 1993

Rice (Oryza saliva L.) calli containing both embryogenic callus (EC) and non embryogenic callus (NEC) regions were initiated from the mature seed on MS medium supplemented with 2.0 mg/L 2,4-D, 0.5 mg/L kinetin. The calli were developed into two callus type which can be distinguished by visual examination depending on color and appearance. In order to illucidate the polypeptide composition between EC and NEC, the total protein extracted from two types of callus was analysed by electrophoresis. By one-dimesional anlaysis of SDS-PAGE and Isoelectric focusing, several protein bands showed quantitative and qualitative difference in each type of callus. The further analysis of the total protein with two-dimensional electrophoresis showed at least 20 EC specific protein and 10 NE specific protein. Also 3 specific protein spots showing micro heterogeneity of 90, 65, 50 kD were detected in EC, while a series of acidic heterologous protein spots were visualized in NEC.in NEC.

• Food Science of Animal Resources
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• v.42 no.5
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• pp.849-860
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• 2022

The myofibril protein (MP) isolate-saccharide graft reactions was prepared using the Maillard reaction with saccharides. The effects of various saccharides on protein functionality and quality of the Maillard reaction were investigated and compared with those of MP. The grafting degree of the MP isolate-saccharide graft reaction was significantly higher in the reducing sugar-treated groups (lactose, glucose, fructose, and palatinose). The browning intensity of the MP isolate-saccharide graft reaction with fructose, sucrose, and erythitol was higher than that observed in the control reaction (p<0.05). MP that reacted with reducing sugars (glucose, fructose, palatinose, and lactose) had fainter bands than MP that reacted with non-reducing sugars (sucrose, erythitol, trehalose, sorbitol, and xylitol). MPs conjugated with glucose exhibited higher protein solubility. The palatinose and lactose treatments were maximum in water binding capacity, though no significant difference in oil binding capacity among the saccharide treatments was observed. The emulsion stability of the MP isolate-saccharide graft reaction with palatinose and erythitol was higher than that of the control reaction. Therefore, reducing sugars have good protein functionality in the MP isolate-saccharides graft reaction.

• Journal of Ginseng Research
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• v.13 no.2
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• pp.158-164
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• 1989

In order to determine the relationships between the lea(-burning disease and the light harvesting chlorophyll-protein (LHCP) complex in Panax ginseng C. A. Meyer, we investigated the chlorophyll-protein (CP) complex of the thylakoid membrane and its characteristics. In P. ginseng four Cp-complex bands determined by non-denaturing SDS-PAGE were identified CP I'(containing reaction center of photosystem I and LHCP I antennae), CP I (reaction center of photosystem I) LHCP II** (oligoform of LHCP II), and LHCP II (photosystem II antennae, CP 26 and CP 29) by Bassis and Dunahay's procedures. Under our experimental condition, the CP I band was only observed in P. ginseng and the band intensity of LHCP II** in P ginseng was higher than in spinach and soybean. There were differences in the absorption and fluorescence spectra and chlorophyll a/b ratio of the CP-complex bands between P. ginseng and other Plants. The Polypeptidr content of P. ginseng thylakoid was lower than in spinach and soybean thylakoid, and the Polypeptide profiles of P. ginseng was low band intensity, especially about 29-35 kD, 55 kD, and 60 kD, compared to spinach and soybean. The inhibitory effects of 2,5-dimethylfuran, specific singlet oxygen ($^1O_2$) quencher, showed that singlet oxygen destroyed 60% of chl.a, 90% of chl.b and 70% of carotenoid in bleaching P. ginseng with leaf-burning disease.

• Korean Journal of Weed Science
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• v.8 no.2
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• pp.199-207
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• 1988

The study was intended to know any relations between the rice tolerance to oxyfluorfen and varietal speciation in seed protein composition or any enzymatical allelies with or without chemical treatment. Rice varieties used were Chokoto, Aichiasahi, Agabyeo, IR 3941 and Tablei as the tolerant group, and Mushakdanti, Weld Pally, HP 1033, HP 857, and HP 907 as the susceptible, respectively. Electrophoretic methods used were SDS-PAGE for seed protein, 7% PAGE for isozymes (acid phosphatase and peroxidase from rice seedling) and changes in isoenzyme activity (malate dehydrogenase, peroxidase and esterase) as affected by oxyfluorfen treatment ($10^{-4}M$) was also studied. The results are summarized as follows. -Among 19 bands separated in seed proteins, two different rice groups selected in terms of tolerance were clustered in dissimilarity. This was based on 2 facts in that G band was not present in susceptible varieties and that less activity of H, N, O, P, Q, Rand S band was shown. -Among 4 bands separated in acid phosphatase, the presence of (band and lower activity of B band was specific for tolerant varieties. For 4 minor bands separated in peroxidase, the tolerant varieties had no activity in B band and higher activity in A, C, D bands. -Time-course study of isozymes as affected by $10^{-4}M$ oxyfluorfen showed that Chokoto, the tolerant varieties, had little activity in A band and consistently higher activities in Band C bands for malate dehydrogenase. For 5 bands separated in peroxidase, B band was not found in Chokoto while A, C, D, and E bands were consistently present. Esterase was separated into about 4 bands in which Chokoto had maintained higher activities in A, C and D bands.

• BMB Reports
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• v.28 no.3
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• pp.197-203
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• 1995

Famesyl protein transferase involved in the first step of post-translational modification of $p21^{ras}$ proteins transfers the famesyl moiety from famesyl pyrophosphate to a cysteine residue in $p21^{ras}$ proteins. The enzyme was first purified 30,000-fold from bovine testis by use of 30~50% ammonium sulfate fractionation, DEAE-Sephacel ion exchange chromatography, Sephacryl S-300 gel filtration chromatography, Sephacryl S-200 gel filtration chromatography, and hexapeptide (Lys-Lys-Cys-Val-Ile-Met) affinity chromatography. The molecular weight of the purified enzyme was estimated to be ~100 kDa by gel filtration and SDS-polyacrylamide gels showed two closely spaced bands of ~50 kDa protein. These indicate that the enzyme consists of two nonidentical subunits, a and 13, which have slightly different molecular weights. The enzyme was inhibited by hexapeptide (Lys-Lys-Cys-Val-Ile-Met), which acted as an alternative substrate that competed for famesylation. Kinetic analysis by measuring initial velocities showed that famesyl protein transferase is a very slow enzyme. EDTA-treated famesyl protein transferase showed little activity with $Mg^{2+}$ or $Zn^{2+}$ alone, but required both $Mg^{2+}$ and $Zn^{2+}$ for the catalytic activity.

• BMB Reports
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• v.39 no.6
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• pp.671-676
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• 2006

The holoenzyme of protein phosphatase (PP) from tulip petals was purified by using hydrophobic interaction, anion exchange and microcystin affinity chromatography to analyze activity towards p-nitrophenyl phosphate (p-NPP). The catalytic subunit of PP was released from its endogenous regulatory subunits by ethanol precipitation and further purified. Both preparations were characterized by immunological and biochemical approaches to be PP2A. On SDS-PAGE, the final purified holoenzyme preparation showed three protein bands estimated at 38, 65, and 75 kDa while the free catalytic subunit preparation showed only the 38 kDa protein. In both preparations, the 38 kDa protein was identified immunologically as the catalytic subunit of PP2A by using a monoclonal antibody against the PP2A catalytic subunit. The final 623- and 748-fold purified holoenzyme and the free catalytic preparations, respectively, exhibited high sensitivity to inhibition by 1 nM okadaic acid when activity was measured with p-NPP. The holoenzyme displayed higher stimulation in the presence of ammonium sulfate than the free catalytic subunit did by protamine, thereby suggesting different enzymatic behaviors.