• Journal of the Korean Society of Food Science and Nutrition
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• v.31 no.2
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• pp.263-270
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• 2002

In the recent studies, many researchers are interested in foods as functional components rather than nutrient sources. Cow's milk is considered as an excellent food sources because of its many nutrients. Casein is a major milk protein and has been reported to have hyperlipidemic and hypercholesterolemic effects. But several reporters have suggested that peptide fractions and hydrolysate of casein have hypolipidemic effects differing from intact protein, casein. Therefore, the objective of the study was to investigate how the casein peptide fractions affect lipid metabolism in rats fed normal or high fat diets. The peptide fractions and hydrolysate of casein were obtained by casein hydrolysis with trypsin. The male rats (Sprague-Dawley), weighing approximately 150 g, were fed each experimental diet containing casein (CAS), casein hydrolysate (CH), casein hydrolysate precipitate (Cpt) and two kinds of peptide fractions (CL & CB) for three weeks, respectively. In the exit I, the male rats were fed normal fat diets (7% soybean oil & cholesterol-free; Expt. I), and in the expt II, fed high fat diets (18% beef tallow & 1% cholesterol; Expt. II). Crude protein contents were calculated from nitrogen contents. Amino acid composition of each fraction was also analyzed. The concentration of total lipid, total cholesterol and triglyceride in serum, liver and feces were measured. As the results of study, tole rats fed peptide fractions with normal fat diets (Expt. I) had no effects on total lipid, total cholesterol and triglyceride concentration in serum and liver and fecal excretion. However, in the rats fed hydrophobic casein peptide fractions (CB) with high fat diet, fecal lipids excretion were significantly increased and the lipids concentration of serum and those of liver tended to decrease, numerically.

• Journal of the Korean Society of Food Science and Nutrition
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• v.15 no.1
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• pp.45-55
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• 1986

This paper aims to study the reactions of lipid or oxidized lipid with protein during drying and storing hair tail fish(Trichurus lepturus) and flounder(Kanakius kitaharai) being generally consumed as dried seafood products in Korea and their influence on the drop of in vitro protein digestibility of these fish meat. The results of the study are as follows: The digestibility of the raw materials of flounder and hair tail fish was 87.63% and 86.08% respectively, and that of sundried and hot air dried materials went down $1{\sim}2$ percent with drying process. But in case of defatted and sundried materials, the rate increased 85.15% and 87.15% respectivley. After 30 days of storage, the digestibility decreased in all materials, and hot air dried meat showed a significant decrease. Trypsin indigestible substrate (TIS) contents of flounder and hair tail fish, in case of raw materials were 0.88 and 0.96mg/g. solid repectiveiy and in case of defatted and sundried materials, TIS contents showed a low increase and digestibility showed a high increase. Brown pigment formation had a wide range of increase in case of the sundried and hot air dried materials and it was increased with duration of storage and temperature. The major fatty acids in the fats of hair tail fish and flounder were $C_{18:1},\;C_{16:0},\;C_{22:6}\;and\;C_{16:1}$ and rate of unsaturated to saturated fatty acids was 79.2:20.8 for flounder, 67.8:32.2 for hair tail fish. After 30 days of storage at room temperature. saturated fatty acids increased compared with the raw materials while unsaturated fatty acids showed a tendency to decrease. Avaialble lysine of hair tail fish was higher than that of flounder and both of them lost about 8.23% of that in raw materials after 30 days of storage.

• The Korean Journal of Zoology
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• v.18 no.2
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• pp.71-86
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• 1975

The G-banding patterns of normal and of delayed spiralized chromosomes by BUdR were investigated in three established cell lines of dwarf hamsters. The results obtained were as follows: 1. The number of G-bands appeared in Chinese hamster T-233 cell line was 65. The centromeric dark band was found in No.1 chromosome and weakly stained bands were also observed in part of the centromeric regions of Nos. 2, 3, 8 and $X_2$ chromosomes. Two homologous X chromosomes were found in different banding patterns. Terminal dark bands were shown in No. 1 chromosome. No conspicuous bands appeared in No. 10 chromosome. 2. Eighty four bands appeared in Armenian hamster Y-1249 cell line. Centromeric dark bands were observed in Nos. 5 and 10 chromosomes and moderatly stained bands were also found in near the centromeric region of the long arms of Nos. 7 and 9 chromosomes. Two isomorphic X chromosomes were also distinguished by their banding patterns. 3. In Y-1313 Armenian hamster cell line, the bands were 69. No centromeric dark bands were observed in this cell line, but moderatly stained bands appeared in the centromeric area in the long arm of No. 9 chromosome. The banding patterns of these two cell lines of Armenian hamster were quite different and readily distinguished. Only No. 8 chromosome showed similar G-banding patterns. Although Nos. 5, 7 abd 8 chromosomes revealed the same number of bands in these two cell lines, the location and staining intensity were quite different. 4. Chromosomes of Nos. 1, 2, 6, $X_1$ and $X_2$ in T-233 cell line and of 1, 4, 7, 8, 9, $X_1$ and $X_2$ in both cell lines of Armenian hamster were found to be elongated due to the inhibition of mitotic spiralization by BUdR. G-banding patterns of these chromosomes were found to be identical to those of normal chromosomes in these cell lines.

• Journal of Life Science
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• v.16 no.4
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• pp.571-578
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• 2006

HtrA2/Omi, a mitochondrial trypsin-like serine protease, is pivotal in regulating apoptotic cell death. Several lines of recent evidence suggest that HtrA2 is associated with the pathogenesis of neurodegenerative disorders; however, the physiological function of HtrA2 still remains elusive. For studying physiological function of HtrA2 in depth, it is necessary to develop a suitable expression system in the model animal. We therefore utilized the zebrafish as a model animal to establish expression of human HtrA2 (hHtrA2) in vivo. For expression of mature HtrA2 as GFP fusion in zebrafish embryos, the HtrA2 (WT) or (S306A) cDNAs with the C-terminal GFP tag were inserted into the pCS2+ plasmid. Expression patterns of HtrA2 in HEK293 cells were first monitored by immunofluorescence staining and immunoblot assays, showing approximately 64 kDa of the HtrA2-GFP fusion proteins. Subsequently, the hHtrA2 plasmid DNA or in vitro transcribed mRNA was microinjected into zebrafish embryos. The expression patterns of HtrA2 in Zebrafish embryos were monitored by GFP fluorescence in 24 hours-post-fertilization (hpf). Although expression patterns of HtrA2-GFP in developing embryos were different between the injected DNA and mRNA, both nucleic acids revealed good expression levels to further study the physiological role of HtrA2 in vivo. This study provides a suitable condition for expressing hHtrA2 in the zebrafish embryos as well as a method for generating useful system to investigate physiological properties of the specific human genes.

• Journal of Life Science
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• v.19 no.7
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• pp.994-1002
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• 2009

A bacteriocin-producing bacterium identified as Bacillus licheniformis was isolated from soybean sauce. Antibacterial activity was confirmed by paper disc diffusion method, using Micrococcus luteus as a test organism. The bacteriocin also showed antibacterial activities against Bacillus sphaericus, Lactobacillus bulgaricus, Lactobacillus planiarum, Paenibacillus polymyxa, and Pediococcus dextrinicus. Optimal culture conditions for the production of bacteriocin was attained by growing the cells in an MRS medium at a pH of 6.5~ 7.0 and a temperature of 37$^\circ$C for 36$\sim$48 hr. Solvents such as chloroform, ethanol, acetone, and acetonitrile had little effect on bacteriocin activity. However, about 50% of bacteriocin activity diminished with treatment of methanol and isopropanol at the final concentration of 50% at 25$^\circ$C for 1 hr. It was stable against a pH variation range from 3.0 and 7.0, but the activity reduced to 50% at a pH range from 9.0 to 11.0. It's activity was not affected by heat treatment at 100$^\circ$C for 30 min and 50% of activity was retained after heat treatment at 100$^\circ$C for 60 min, showing high thermostability. The bacteriocin was purified to a homogeneity through ammonium sulfate precipitation, SP-Sepharose ion-exchange chromatography, and reverse-phase high-performance liquid chromatography (HPLC). The entire purification protocol led to a 75-fold increase in specific activity and a 13.5% yield of bacteriocin activity. The molecular weight of purified bacteriocin was estimated to be about 2.5 kDa by tricine-SDS-PAGE.

• Journal of Embryo Transfer
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• v.18 no.3
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• pp.171-178
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• 2003

This study was conducted to investigate the effect of electric fusion methods on cell fusion rate and embryo development by somatic cell nuclear transfer in Korean Native Cattle. The KNC ear cell was cultured in vitro for confluence in serum starvation condition(DMEM＋0.05％ FBS) for cell confluence. The zona pellucida of IVM oocytes were partially dissection using micro pipette. Ear cells were transferred into an enucleated oocyte. The reconstructed embryos were electrically fused with Zimmermann Cell Fusion Medium(ZCFM). Nuclear transfer embryos were activated with a combination of 10${\mu}{\textrm}{m}$ calcium ionophore(5 min) and 2.0mM 6-DMAP(3 hr). The activated embryos were cultured in CR1 -aa medium contains 0.3％ BSA or 10％ FBS at 37$^{\circ}C$, 90％ $N_2$, and 5％ $CO_2$in incubator for 6 days. The fusion rates were 51.6％(chamber) and 68.9％(needle), respectively and there were significantly difference between the fusion method(P<0.05). But, lysis rates were not significantly different(10.7％, 11.5％), respectively. The cleavage rates were significantly different between the chamber method(73.2％) and needle method(80.3％), respectively(P<0.05). The rates of early embryos(2∼4cells) and blastocysts of chamber and needle methods were 54.1％, 61.1％ and 18.4％, 26.3％ respectively, and needle method was significantly higher than chamber method(P<0.05). But, morulae formation rate were not significantly differences between the chamber(6.7％) and needle(6.2) method(P <0.05). These result suggest that electric fusion of needle method was to be profitable for nuclear transfer embryo fusion rate, blastocyst formation rate and reduce of oocyte lysis.

• Korean Journal of Animal Reproduction
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• v.26 no.2
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• pp.125-132
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• 2002

This study was conducted to examine the effects of electric stimulation conditions on in vitro developmental ability of procine embryos after somatic cell nuclear transfer, The porcine ear cell was cultured in vifro for confluency in serum-starvation condition (TCM-199＋0.5% FBS) for cell confluency. The zona pellucida of IVM oocytes were partially drilled using laser system. Single somatic cell was individually transferred into the enucleated oocyte. The reconstructed embryos were electrically fused with 0.3M mannitol. After electric fusion, the embryos were activated and cultured in NCSU-23 medium containing 10% FBS at 39$^{\circ}C$, 5% $CO_2$ in air for 6 to 8 days. Nuclear transferred(NT) oocytes which fused at a field strength of 1.90kv/cm showed a higher (P<0.05) fusion rate(49.5%, 50/101) compared to 2.10 kv/cm(25.8%, 24/93) or 2.50kv/cm(30.3%, 27/89). After electric activation, the cleavage rate of NT embryos was 48.0(24/50), 66.6(16/24) and 70.3% (19/27), respectively and these were not different. There was no significant difference in fusion rate by duration and pulse of electric stimulation. In cleavage rate, however, more NT embryos(76.3%, 45/59) cleaved at 60 $\mu$sec twice than other embryos(49.1 to 56.5%) with different conditions of electric stimulation(P<0.05). NT embryos activated at a field strength of 1.50kv/cm showed a higher developmental rate(9.8%, 5/51) than those embryos activated at 1.25kv/cm(0%) or parthenotes(6.4%, 7/109). These results suggest that some factors such as field strength, duration and pulse of electric stimulation could be affected to in vitro developmental ability of nuclear transplanted porcine embryos.

• Journal of Plant Biotechnology
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• v.29 no.3
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• pp.161-170
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• 2002

For the hairy root of Panax ginseng, we have got mass spectrums from MALDI/TOF/MS analysis and Tandem mass spectrums from ESI/Q-TOF/MS analysis. While mass spectrum provides the molecular weights of peptide fragments digested by protease such as trypsin, tandem mass spectrum produces amino acid sequence of digested peptides. Each amino acid sequences can be a query sequence in BLAST search to identify proteins. For the specimens of animals or plants of which genome sequences were known, we can easily identify expressed proteins from mass spectrums with high accuracy. However, for the other specimens such as ginseng, it is difficult to identify proteins with accuracy since all the protein sequences are not available yet. Here we compared the mass spectrums and the peptide amino acid sequences with ginseng expressed sequence tag (EST) DB. The matched EST sequence was used as a query in BLAST search for protein identification. They could offer the correct protein information by the sequence alignment with EST sequences. 90% of peptide sequences of ESI/Q-TOF/MS are matched with EST sequences. Comparing 68% matches of the same sequences with the nr database of NCBI, we got more matches by 22% from ginseng EST sequence search. In case of peptide mass fingerprinting from MALDI/TOF/MS, only about 19% (9 proteins of 47 spots) among peptide matches from nr DB were correlated with ginseng EST DB. From these results, we suggest that amino acid sequencing using tandem mass spectrum analysis may be necessary for protein identification in ginseng proteome analysis.

• Journal of Life Science
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• v.28 no.11
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• pp.1339-1346
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• 2018

In Korea, edible mushrooms are produced largely on commercial artificial media, so the annual production of spent mushroom substrate (SMS), as a by-product of the mushroom industry, is estimated at over 200 million tons. This SMS is assumed to contain abundant fungal mycelia and pre-fruiting bodies, as well as various nutritive and bioactive compounds that are presently discarded. This study examined the physico-chemical, nutritional, and enzymatic characteristics of uninoculated sterilized medium (USM) and SMS of shiitake mushrooms with the aim of developing a high-value added product from SMS. The contents of crude protein, crude lipid, and ash were higher after the third SMS harvest ($SMS-A-3^{rd}$) than in USM or $SMS-A-1^{st}$. The contents of Ca, Mg, and P in $SMS-A-3^{rd}$ were 2.95, 2.35, and 2.1-fold higher compared than in USM. No As or Cd was detected in USM or SMS. The pH, Brix, and acidity were 4.6, 20.0, and 1.4, respectively in $SMS-A-3^{rd}$, but 5.6, 6.0, and 0.0, respectively, in USM. These results suggest a highly active production of soluble components and organic acids in $SMS-A-3^{rd}$. The distinct color differences noted for USM, $SMS-A-1^{st}$, and $SMS-A-3^{rd}$ could be used as a mycelial growth indicator. Enzyme activity assays using the APIZYM system showed that SMS is a potent source of hydrolysis-related enzymes, especially esterase (C4) and ${\beta}$-glucuronidase. Our results suggested that the SMS of shiitake has a high potential for use in environmental, agricultural, and stock-breeding industries, for example, as active ingredients for sewage treatment, waste-polymer degradation, and feed additives.

• Journal of Life Science
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• v.29 no.8
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• pp.895-903
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• 2019

The aim of the present study was to improve the cell viability of human dental papilla derived single-induced pluripotent stem cells (iPSCs) using a Rho-associated protein kinase (ROCK) inhibitor, Y-27632. The iPSCs were produced using an episomal plasmid-based reprogramming method. After cell separation using trypsin, the iPSCs were treated with 0, 0.5, 1, 2.5, 5, 7.5, or $10{\mu}M$ Y-27632 for 5 d. Cell viability increased significantly following the $5{\mu}M$ Y-27632 treatment (p<0.05). When the iPSCs were exposed to medium containing $10{\mu}M$ Y-27632 for 0, 1, 2, 3, 4, and 5 d, the cell viability rate increased significantly in accordance with the cell viability rate (p<0.05). To evaluate the effect of the Y-27632 treatment on stemness characteristics, the expression of stem cell-specific transcripts and telomerase activity were investigated in the iPSCs treated with $10{\mu}M$ Y-27632 for 5 d. The expression levels of stem cell-specific transcripts, such as OCT-4, NONOG, and SOX-2, and telomerase activity were not significantly different in the iPSCs treated with $10{\mu}M$ Y-27632 as compared with those of untreated control iPSCs (p>0.05). Taken together, the results demonstrated that cell viability can be improved by treatment with the ROCK inhibitor Y-27632, without losing iPSC stemness characteristics.