• Korean Journal of Food Science and Technology
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• v.30 no.5
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• pp.1236-1242
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• 1998

Korean Lentinus edodes SR-1 was cultured to multiply the mycelia in the complete broth medium (C/N=13.1) for mushroom, and protein-bound polysaccharides were extracted from the cultured broth containing mycelia (The whole cultured broth was used to increase the yields: 80% of protein-bound polysaccharides were existed at the cell wall of mycelia and 20% of those were secreted extracellularily in this culture). Protein-bound polysaccharides in the cultured broth containing mycelia were extracted by using three different methods: 1) Extraction with hot water, 2) Disintegration of cell wall by glass bead mill treatment before extraction with hot water, and 3) Cellulase treatment before extraction with hot water. The highest yield was obtained (930 mg polysaccharides/100 mL culture broth) when protein-bound polysaccharides were extracted with 2) method. The extracted crude protein-bound polysaccharides were purified using protease, DEAE-cellulose and Sephadex G-100. The growth inhibition activity for $P_{388}$, mouse leukemic cell, increased (53.7, 62.2, 93.7% and 97.4%) as the purification level increased. Protein-bound polysaccharides contained 46.1% of polysaccharides, 7.3% of protein, and trace amounts of minerals. Polysaccharides contained glucose, galactose, xylose and mannose. The content of proline and glycine were high, however, methionine and leucine were not found. The major minerals were Na, K, Zn, and Ca.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.6
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• pp.2447-2451
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• 2014

Human papillomavirus (HPV) is the primary etiologic agent of cervical cancer. Consideration of safety and non human leukocyte antigen restriction, protein vaccine has become the most likely form of HPV therapeutic vaccine, although none have so far been reported as effective. Since tumor cells consistently express the two proteins E6 and E7, most therapeutic vaccines target one or both of them. In this study, we fabricated DC vaccines by transducing replication-defective recombinant adenoviruses expressing E6/E7 fusion gene of HPV-16, to investigate the lethal effects of specific cytotoxic T lymphocytes (CTL) against CaSki cells in vitro. Mouse immature dendritic cells (DC) were generated from bone marrow, and transfected with pAd-E6/E7 to prepare a DC vaccine and to induce specific CTL. The surface expression of CD40, CD68, MHC II and CD11c was assessed by flow cytometry (FCM), and the lethal effects of CTL against CaSki cells were determined by DAPI, FCM and CCK-8 methods. Immature mouse DC was successfully transfected by pAd-E6/E7 in vitro, and the transfecting efficiency was 40%-50%. A DC vaccine was successfully prepared and was used to induce specific CTL. Experimental results showed that the percentage of apoptosis and killing rate of CaSki cells were significantly increased by coculturing with the specific CTL (p <0.05). These results illustrated that a DC vaccine modified by HPV-16 E6/E7 gene can induce apoptosis of CaSki cells by inducing CTL, which may be used as a new strategy for biological treatment of cervical cancer.

• Applied Biological Chemistry
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• v.39 no.3
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• pp.177-182
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• 1996

Effects of 14 carbohydrates supplied as carbon sources on cell growth and sporulation of, and the production of insecticidal crystal proteins by Bacillus thuringiensis strains were investigated in liquid cultures. Strains grew well in media containing any one of the 14 carbohydrates supplied, reaching maximum cell densities of $10^7{\sim}10^8\;cells/ml$ in 16.7 to 22 hours after inoculation depending on the strain. Spores first appeared in 16.7 to 24.7 hours after inoculation, and 80% sporulation was reached in 28 to 51.3 hours after inoculation depending on the strain. No change in pH of media was observed after cell multiplication. The production of total protein was highest when supplied with sucrose and was lowest with starch. More insecticidal crystal proteins were produced when supplied with glucose, lactose, maltose, or sucrose. The amount of insecticidal crystal proteins produced by the strains was proportional to that of the total protein. The relative amount of individual insecticidal crystal protein species produced by B.t. kurstaki and B.t. israelensis was not influenced by the carbohydrates supplied.

• The Korean Journal of Zoology
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• v.15 no.2
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• pp.92-93
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• 1972

Ox-RBC로부터 Moddy法에 依하여 Ox-RCM抗原을 抽出하여 몇가지 分析을 實施하였다. Lyophilized 한 Ox-RCM protein의 化學的 分析 結果 total protein은 $55\\pm 1.3%$, total nitrogen은 9.2%, carbohydrate 8.5% 그리고 total acid soluble phosphorus는 $0.96\\pm 0.1%$였다. Polyacrylamide gel electrophoresis한 結果 Ox-RCM은 12 $\\sim$ 15個, Pre-VRS fraction은 8個 및 PHR 分劃은 9個의 pattern을 나타냈다. Amino acid의 分析 結果로 cystine이 4.93 $\\sim$ 3.72 moles/100 moles 이고 glycine이 最高로 26.7 $\\sim$ 13.5 moles/100 moles였고 最低로 methionine 이었으나 試料의 抽出方法에 따라 그 値의 變動을 볼 수 있다.

• The Korean Journal of Food And Nutrition
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• v.24 no.4
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• pp.740-745
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• 2011

Ohmic heating uses electric resistance heat which occurs equally and rapidly inside food when an electrical current is passed throught. In this study, we observed the physical & chemical characteristics changes which occurs in soybean protein during heating denaturation by using ohmic and conventional heating. After the ohmic heating process, we could not find any change of the primary protein structure in the denaturated soy protein samples. However, the rate of imbibed water(RIW) of the ohmic samples was 2 times faster than that of the conventional samples. Also the ANS-surface hydrophobicity was decreased, which is very closely related to RIW. In the differential scanning calorimeter(DSC) analysis result, all 7S soyprotein fraction samples were completely denaturated by ohmic and conventional heating. However, the 11S samples were completely denatured only by ohmic heating. According to the DSC result, we decided that soyprotein was damaged by temperature and electrical current during ohmic heating. The damage of electrical current was a cause of the characteristic changes.

• Journal of the Korean Home Economics Association
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• v.7 no.1
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• pp.63-68
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• 1969

• BMB Reports
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• v.33 no.3
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• pp.242-248
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• 2000

Phage display of proteins is a powerful tool for protein engineering since a vast library of sequences can be rapidly screened for a specific property. In this study, we develop da new phage display vector that was derived from a pET-25b(+) vector. The pET-25b(+) was modified in order that the expressed protein would have a T7-tag at the amino terminus and GpS (a major coat protein of M13 phage) at the carboxyl terminus. Another vector without the gp8 gene was also constructed. The newly developed phagemid vectors have several advantageous features. First, it is easy to examine whether or not the target proteins are functional and faithfully transported into the periplasmic space. This feature is due to the fact that recombinant proteins are produced abundantly in the pET system. Second, the T7-tag makes it possible to detect any target proteins that are displayed on the surface of filamentous bacteriophage. To verify the utility of the vector, the clones containing the glutathione S-transferase (GST) gene as a target were examined. The result showed that the GST produced from the recombinant vector was successfully transported into the periplasmic space and had the anticipated enzyme activity. Western blot analysis using a T7-tag antibody also showed the presence of the target protein displayed on the surface of the phage. The phages prepared from the recombinant clones were able to bind to glutathione-Sepharose and then eluted with glutathione. These results showed that the new vectors developed in this study are useful for the phage display of proteins.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.5
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• pp.712-718
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• 2004

This study was carried out to separate the calcium-binding protein derived from enzymatic hydrolysates of cheese whey protein. CWPs (cheese whey protein) heated for 10 min at $100^{\circ}C$ were hydrolyzed by trypsin, papain W-40, protease S, neutrase 1.5 and pepsin, and then properties of hydrolysates, separation of calcium-binding protein and analysis of calcium-binding ability were investigated. The DH (degree of hydrolysis) and NPN (non protein nitrogen) of heated-CWP hydrolysates by commercial enzymes were higher in trypsin than those of other commercial enzymes. In the result of SDS-PAGE (sodium dodecyl sulphate polyacrylamide gel electrophoresis), $\beta$-LG and $\alpha$-LA in trypsin hydrolysates were almost eliminated and the molecular weight of peptides derived from trypsin hydrolysates were smaller than 7 kDa. In the RP-HPLC (reverse phase HPLC) analysis, $\alpha$-LA was mostly eliminated, but $\beta$-LG was not affected by heat treatment and the RP-HPLC patterns of trypsin hydrolysates were similar to those of SDS-PAGE. In ion exchange chromatography, trypsin hydrolysates were shown to peak from 0.25 M NaCl and 0.5 M NaCl, and calcium-binding ability is associated with the large peak, which was eluted at a 0.25 M NaCl gradient concentration. Based on the results of this experiment, heated-CWP hydrolysates by trypsin were shown to have calcium-binding ability.

• Preventive Nutrition and Food Science
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• v.7 no.2
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• pp.139-145
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• 2002

Hen egg-white lysozyme, ovalbumin, egg-yolk phosvitin, acid-precipitated soy protein and $\alpha$$_{sl}$ milk casein were covalently linked with galactomannan through a controlled dry-heating at 6$0^{\circ}C$ under 79% relative humidity without any chemical reagent. Neoglycosylation by the covalent binding of polysaccharide chains brought a significant improvement into the surface functionalities of food proteins. Excellent emulsifying properties and foaming properties were observed in all protein-galactomannan conjugates. Bacterial mutagenesis tests and animal dose test were done to evaluate the food safety of the protein-galactomannan conjugates. The neo-glycoproteins were negative for Ames test using Salmonella typhimurium TA100 (hisG46) and TA98 (hisD3052) strains, and rec-assay using Bacillus subtilis Hl7 (rec) and M45 (re $c^{+}$) strains. All substances were also nontoxic for oral administration to rats. L $D_{50}$ 's of these substances were all more than 7.5 g/kg body-weight of rat. No effect was also observed in the weight increases and the concentrations of total cholesterol, triglyceride and phospholipids in blood serum of the administrated rats with 7.5 g/kg conjugates. Thus, Maillard-type protein-polysaccharide conjugates prepared by covalent attachment of galactomannan to food proteins were proposed to be useful as a safe functional biopolymer in this study.y.

• Journal of Crop Science and Biotechnology
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• v.10 no.4
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• pp.231-236
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• 2007

Differentially expressed genes(DEG) were identified in a rice variety, Sathi, an indica type showing high allelopathic potential against barnyardgrass(Echinochloa crus-galli(L.) Beauv. var. frumentaceae). Rice plants were grown with and without barnyardgrass and total RNA was extracted from rice leaves at 45 days after seeding. DEG full-screening was performed by $GeneFishing^{TM}$ method. The differentially expressed bands were re-amplified and sequenced, then analyzed by Basic Local Alignment Search Tool(BLAST) searching for homology sequence identification. Gel electrophoresis showed nine possible genes associated with allelopathic potential in Sathi, six genes(namely DEG-1, 4, 5, 7, 8, and 9) showed higher expression, and three genes(DEG-2, 3 and 6) showed lower expression as compared to the control. cDNA sequence analysis showed that DEG-7 and DEG-9 had the same sequence. From RT PCR results, DEG-6 and DEG-7 were considered as true DEG, whereas DEG-1, 2, 3, 4, 5, and 8 were considered as putative DEG. Results from blast-n and blast-x search suggested that DEG-1 is homologous to a gene for S-adenosylmethionine synthetase, DEG-2 is homologous to a chloroplast gene for ribulose 1,5-bisphosphate carboxylase large subunit, DEG-8 is homologous to oxysterol-binding protein with an 85.7% sequence similarity, DEG-5 is homologous to histone 2B protein with a 47.9% sequence similarity, DEG-6 is homologous to nicotineamine aminotransferase with a 33.1% sequence similarity, DEG-3 has 98.8% similarity with nucleotides sequence that has 33.1% similarity with oxygen evolving complex protein in photosystem II, DEG-7 is homologous to nucleotides sequence that may relate with putative serin/threonine protein kinase and putative transposable element, and DEG-4 has 98.8% similarity with nucleotides sequence for an unknown protein.