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Purification and Characterization of Alkaline Protease from saewoo-jeot, salted and fermented shrimp (Acetes japonicus) (새우젓에서 alkaline pretease의 정제 및 특성)

  • Nam, Eun-Jung;Oh, Se-Wook;Jo, Jin-Ho;Kim, Young-Myung;Yang, Cha-Bum
    • Korean Journal of Food Science and Technology
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    • v.30 no.1
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    • pp.82-89
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    • 1998
  • This study was performed to elucidate the purification and characterization of pretease from saewoo-jeot, a Korean traditional salt-fermented shrimp product. The protease in saewoo-jeot (Acetes japonicus) were extracted, desalted through electrodialysis and purified by ammonium sulfate fractionation, Sephadex G-100 gel filtration and DEAE-cellulose column chromatography. Purified enzyme had specific activity of 8.4 unit/mg, yield of 14% and purification fold of 9.8. Purified enzyme was confirmed as single band protein by polyacrylamide gel electrophresis and the molecular weight was estimated to be about 24 kDa. The optimal pH and temperature for the enzyme activity were 8.0 and $40^{\circ}C$, respectively. The range of its stability to the pH and temperature were 7.0 to 10.0 and $30^{\circ}C\;to\;60^{\circ}C$, respectively. The activity of enzyme to synthetic substrate showed BAPNA and TAME. The enzyme was activated significantly by manganese ions, while inhibited by STI, TLCK. metals $(K^+,\;Li^+,\;Na^+,\;Ca^{++},\;Co^{++},\;Cu^{++},\;Mg^{++},\;Ba^{++},\;Hg^{++},\;Zn^{++},\;Fe^{+++})$. The Km value of the enzyme was $5.1{\times}10^{-7}\;M$ to hammersten casein. It's suggested that purified protease from saewoo-jeot seemed to be trypsin-like enzyme.

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Batch Scale Storage of Sprouting Foods by Irradiation Combined with Natural Low Temperature -V. Cooking Qualities of Irradiated Chestnut after Long-term Storage- (방사선조사(放射線照射)와 자연저온(自然低溫)에 의한 발아식품(發芽食品)의 Batch Scale저장(貯藏)에 관(關)한 연구(硏究) -제5보(第五報) : 장기저장(長期貯藏)된 조사(照射)밤의 조리적성(調理適性)에 대하여-)

  • Yang, Ho-Sook;Kim, Jong-Gun;Cho, Han-Ok;Byun, Myung-Woo;Kwon, Joong-Ho
    • Korean Journal of Food Science and Technology
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    • v.15 no.3
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    • pp.238-244
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    • 1983
  • The cooking quality of irradiated chestnut after longterm storage were evaluated. 1. Color degree and tannin content of irradiated chestnut were slightly increased with the storage, but there was a little difference according to the radiation dose after nine months storage. 2. The main component of free sugars in the irradiated chestnut were identified as sucrose, glucose, fructose and the amino acids of chestnut were identified in the decreasing order of glutamic acid, aspartic acid, leucine, arginine, glycine, alanine, serine, pheylalanine, threonine, valine, isoleucine, tyrosine, methionine and cystein. Free sugars and amino acids of 25 Krad irradiated group showed a little difference compared with those of control group after nine months storage. 3. The calorie of candied chestnut prepared from nine months stored was marked 199 Kcal/100g of edible parts compared with 159 Kcal of raw chestnut. 4. Texture and sensory evaluation of candied chestnut prepared from nine months stored were better in 20-25 Krad irradiated group than in control group.

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Fractionation of Rice Bran Lipid and Storage Effects on Bran Lipid Composition (미강(米糠)의 지질성분(脂質成分) 및 저장(貯藏)중 지질특성(脂質特性) 변화(變化)에 관한 연구(硏究))

  • Ryu, Chung-Hee;Cheigh, Hong-Sik
    • Korean Journal of Food Science and Technology
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    • v.12 no.4
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    • pp.278-284
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    • 1980
  • Lipids from rice bran (Indica type Milyang #23), both fresh and stored at $30^{\circ}C$ and 80% relative humidity for 5 weeks, were separated and analyzed for the determination and the storage effect on the bran lipid composition. Total lipids of fresh rice bran consisted of 89.9% neutral lipids, 8.0% glycolipids, 2.1% phospholipids and no significant changes of these fractions were noted during storage. Triglycerides(43.1%), diglycerides(13.8%) and hydrocarbon-esterified sterol(13.5%) among six fractions were considered as main components in neutral lipids. After storage triglycerides content significantly decreased as the free fatty acid increased in the neutral lipid fraction. Major components of the glycolipid fraction were acylsterolglycoside(43.1%) and sterolglycoside(30.3%). Phosphatidyl choline(39.8%), phosphatidyl serine(20.9%) and phosphatidyl ethanolamine(19.8%) were predominent in the phospholipid fraction. No significant changes of the composition were shown in fraction of the glycolipid or the phospholipid during the storage period. Major fatty acids of the total lipid fraction were oleic(44.3%), linoleic(32.5%) and palmitic acids(18.4%). The fatty acid compositions of the neutral lipid, the glycolipid and the phospholipid fractions were similar to the total lipid fraction. Small changes in fatty acid composition in each fraction were noted during the storage period. The acid value increased but iodine value decreased during the storage period. The values of peroxide and TBA increased gradually in the first three weeks, and then slowly decreased in the fourth and the fifth week of the storage.

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Studies on the Extracts Preparation of Korean Shiitake Mushroom(Lentinus edodes) (한국산 표고버섯 Extract 제조에 대한 연구)

  • Chang, Young-Sang;Lee, Ho-Bong;Lee, Seung-Ryeol;Shin, Zae-Ik
    • Korean Journal of Food Science and Technology
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    • v.22 no.7
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    • pp.828-832
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    • 1990
  • Pileus and stipe extracts of shiitake mushroom were prepared with various ethanol concentration by different extraction time at $60^{\circ}C$. Yields, total reducing sugars, free amino acids and nucleotides in ultrafiltrated extracts were analyzed. Yields were higher in hot water extracts but there was no difference depending on changes of extraction time. Total reducing sugar contents got higher by hot water extraction than by solvent extraction. In hot water extracts of the pileus and stipe, reducing sugar content were in the range of $416.18{\sim}488.18\;mg%\;and\;435.37{\sim}452.12\;mg%$, respectively. Threonine+serine, glutamic acid, lysine and arginine were dominent in the free amino acids pool of raw material. The contents of free amino acids in hot water extracts of pileus and stipe were about 528.46mg% in 2 hr and 221.01 mg% in 3 hr. The proportion of bitter amino acids in extracts to total free amino acid contents was in the range of $16{\sim}29%,\;35{\sim}37%$ in pileus and stipe extracts, respectively. Nucleotides contents were higher in pileus than in the stipe. When the 25% ethanol solution was used for extraction solvent, nucleotides contents in pileus and stipe extracts was high.

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FACTORS INVOLVED IN THE QUALITY RETENTION OF CULTURED UNDARIA PINNATIFIDA (양식미역의 품질요인과 그 가공)

  • PYEUN Jae-Hyeung;PARK Yenog-Ho;LEE Kang-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.10 no.2
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    • pp.125-135
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    • 1977
  • Recently, culture of Undaria pinnatifida, one of the representative esculent sea weed, has been prevailing in tile east and south coasta of Korea and reached the mass culture stage. In this study, compositional quality factors for food were studied and the contributory effects of blanching and pigment fixatives in the quality retention of cultured Undaria pinnatifida are discussed. When the place and time of harvesting were the same, cultured pinnatifida showed scarce difference in the chemical composition comparing to tile naturally grown Undaria pinnatifida, but cultured Undaria pinnatifida shelved a considerable difference depending upon the cultured places. In the chemical composition of Undaria pinnatifida, the alginic acid comprising about $40\%$ of the whole solid materials seemed to be responsible for the compositional puality. The chlorophyll and carotenoid content of the clutured Unaria pinnatifida were considerably lower than that of the naturally grown Undaria pinnatifida and wass inferior in puality by color to the naturally grown one. Dried Undaria pinnatifida contained a considerable amount of amino-N, mannitol, and soluble minerals and it is considered that these components play a great role in the relish effects. It could also be evaluated as a good albuminous source for food science the dried pinnatifida contains about $18\%$ of crude protein. In the analysis of free amino acid composition of dried Undaria pinnatifida, the naturally growm samples showed so what higher levels in all amino acid content than the cultured samples. The contents of theronine, alanine, and glutamic acid were major in quantity wherease histidine cysteine, tyrosine, and phenylalanine were minor. The contents of such amino acids like serine and proline were particularly low or undetectable. The results of amino acid analysis of the acid hydrolysates of dried Undaria pinnatifida in quantity of individual amino acid showed te same pattern as that of free amino acid. It is noticed that Undaria pinnatifida seemed to contain good quality protein since the contents of essential amino acids were considerably higher and uniform. By blanching the fresh sample, the water soluble components brought about cousiderable loss, and, particularly, it was noteworthy that both mannitol and soluble minerals apparently decreased. In the pigment analysis of the dried sample, blanching was effective to retain chlorophyll and carotenoid. The addition of pigment fixatives in blanching solution such as Ca-gluconate, Ca-carbonate, and Ca-hydroxide did not exhibit much effect on the pigment retention except that Ca-carbonate shelved some effect only in the early stage of storage.

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Effects of extruded pellet and moist pellet on growth performance, body composition, and hematology of juvenile olive flounder, Paralichthys olivaceus

  • Lee, Seunghan;Moniruzzaman, Mohammad;Bae, Jinho;Seong, Minj;Song, Yu-jin;Dosanjh, Bakshish;Bai, Sungchul C.
    • Fisheries and Aquatic Sciences
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    • v.19 no.8
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    • pp.32.1-32.6
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    • 2016
  • A feeding trial was conducted to evaluate the effects of two different sizes of extruded pellets (EP) ($EP_1$ - 3 mm or $EP_2$ - 5 mm) and a moist pellet (MP) in olive flounder, Paralichthys olivaceus, reared in semi-recirculation system. A total of 450 fish with an average initial weight of $5.0{\pm}0.2g$ (mean ${\pm}$ SD) were fed one of the three experimental diets in triplicate groups. At the end of a 6-week feeding trial, weight gain, specific growth rate, and feed efficiency of fish fed EP diets were significantly higher than those of fish fed MP (P < 0.05). Water quality parameters like turbidity, total ammonia nitrogen, and total phosphorous from tanks of fish fed $EP_1$ and $EP_2$ were significantly lower than those from tanks of fish fed MP. Blood plasma glutamic oxaloacetic transaminase and glucose concentration were significantly higher in fish fed MP diet compared to fish fed EP diets (P < 0.05). Whole body crude protein contents in fish fed EP diets were higher than those from the fish fed MP diet. Whole body amino acid content like threonine, aspartic acid, serine, tyrosine, and cystine were found to be significantly higher in fish fed EP diets than those in fish fed MP diet. In considering overall performance of olive flounder, $EP_2$ diet could be recommended for the successful aquaculture of this important fish species.

Protein Expression of Matrix Metalloproteinases of Mouse Reproductive Organs During Estrous Cycle (생식주기에 따른 자성 생쥐의 생식기관의 Matrix Metalloproteinase의 단백질 발현)

  • Kim, Moon-Young;Lee, Ki-Won;Kim, Hae-Kwon;Kim, Moon-Kyoo;Cho, Dong-Jae
    • Clinical and Experimental Reproductive Medicine
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    • v.25 no.2
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    • pp.161-170
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    • 1998
  • Protein expression patterns of matrix metalloproteinases (MMPs) were examined in mouse reproductive organs during estrous cycle. Estrous cycle was classified into diestrus, proestrus, estrus or metestus and MMP expression was analyzed by zymography using gelatin as a substrate. Uterine fluid (UF) obtained both at diestrus and proestrus exhibited 4 major MMPs including 106kDa, 64kDa, 62kDa and 59kDa gelatinases. However, in UF at estrus, the gelatinolytic activity of 64kDa MMP disappeared and that of 106kDa and 62kDa MMPs dramatically decreased. At metestrus, 64kDa MMP activity reappeared and 106kDa and 62kDa MMP exhibited increased activities such that the band intensity of 106kDa was comparable to that in UF at diestrus. Gelatinolytic activity of 59kDa MMP was not changed throughout the cycle. Both ovarian and oviductal tissue homogenate revealed 4 MMPs which corresponded to the 4 MMPs of UF. However, unlike UF MMPs, gelatinolytic activity of these MMPs did not show distinct changes throughout the cycle. Either an inhibitor of MMP, 1,10-phenanthroline, or a metal chelator, EDTA, abolished the appearance of the above MMP activities in gelatinated gel whereas a serine proteinase inhibitor, phcnylmethylsulfonyl fluoride, failed to inhibit the appearance of MMP activities, proving that gelatinolytic activity of the above reproductive tissues were due to the enzymatic activity of MMP. When gclatinolytic activity of mouse serum was examined, it revealed 5 MMPs (131kDa, 106kDa, 89kDa, 64kDa and 62kDa bands) and one gelatinase (84kDa) band. From these results, it is concluded that the protein expression of MMPs of mouse reproductive organs, particularly uterus, is temporally regulated during estrous cycle and uterine 106kDa, 64kDa and 62kDa MMPs are suggested to play an important role in cyclic tissue remodeling of mouse uterus.

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Characteristics of Polysaccharide Extracted from the Cultured Mycelia of Schizophyllum commune (치마버섯 균사체 배양물로부터 분리한 다당류의 이화학적 특성)

  • Lee, June-Woo;Kim, Yong-Seok
    • The Korean Journal of Mycology
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    • v.37 no.1
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    • pp.91-95
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    • 2009
  • To examine physico-chemical properties of the polysaccharide extracted from liquid-cultured mycelia of Schizophyllum commune, each the polysaccharide was extracted with hot water treatment and then fractionated with ethanol, alkaline solution and ultrafiltration. And we determined carbohydrate contents, composition of amino acids, infra-red spectrum and viscosity. Carbohydrate contents of polysaccharide treated with ethanol and ultrafiltration were 72.0% and 62.3%, and proteins content were 15.3% and 32.0% respectively. The carbohydrate consisted of four monosaccharides and the protein contained 16 amino acids. The polysaccharide obtained from ultrafiltration was shown an absorption band characteristic of the ${\beta}$-glycosidic linkage by infra red spectra. These results suggest that the polysaccharide extracted from Schizophyllum commune showed the characteristics of proteinbounded polysaccharide, and it was ${\beta}$-glycosidic linkage with strong viscosity.

Differential Effects of Fumonisin $B_1$ on Cell Death in Cultured Cells: the Significance of the Elevated Sphinganine

  • Yu, Chang-Hun;Lee, Yong-Moon;Yun, Yeo-Pyo;Yoo, Hwan-Soo
    • Archives of Pharmacal Research
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    • v.24 no.2
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    • pp.136-143
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    • 2001
  • Fumonisins are specific inhibitors of ceramide synthase in sphingolipid metabolism. An alteration in sphingolipid metabolism as a result of fumonisin exposure is related to cell death (Yoo et al., 1992). The objective of this study was to investigate whether elevated free sphinganine levels are related to the sensitivity of cultured cells to fumonisin exposure. Fumonisin $B_1$ elevated the intracellular free sphinganine concentraions in both LLC-$PK_1$ and Chinese hamster ovary (CHO) cells. However, CHO cells are resistant to fumonisin cytotoxicity at 50${u}m$, while LLC-$PK_1$ cells are sensitive at concentrations greater than 357M. The intracellular concentration of free sphinganine in LLC-$PK_1$ cells treated at 50${u}m$ fumonisin $B_1$ for 72 h was approximately 1450 pmol/mg protein relative to the 37 pmol observed in the control culture. Under the same conditions, the population of apoptotic cells in the 50${u}m$ fumonisin $B_1$-treated culture was approximately 37% of the total compared to 12% in the control. The caspase III-like activity after 72 h in the 50${\mu}$M fumonisin $B_1$-exposed culture Increased to approximately 50 $pmol/mg$ protein/hr compared to 6 $pmol/mg$ protein/hr in the control. L-cycloserine, a serine palmitoyltransferase inhibitory reduced the fumonisin $B_1$-stimulated caspase III-like activity down to the control level. Under the same culture conditions, the intracellular concentration of free sphinganine after-cycloserine plus fumonisin $B_1$ treatment was 140 pmol/mg protein compared to 1450 $pmol/mg$ protein in fumonisin $B_1$ alone. The intracellular concentration of free sphinganine in CHO cells treated with 50${u}m$ fumonisin $B_1$ for 72 h was al)proximately 460 pmol/mg protein, indicating that the mass amount of elevated free sphinganine in the CHO cells was about 32% of that in LLC-$PK_1$ cells. Adding exogenous sphinganine to the CHO cells along with 50${u}m$ fumonisin $B_1$ treatment for 72 h caused both necrosis and apoptosis. In conclusion, the elevated endogenous sphinganine acts as a contributing factor to the fumonisin-induced cell death.

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Constitutive Activation of $p70^{S6k}$ in Cancer Cells

  • Kwon, Hyoung-Keun;Bae, Gyu-Un;Yoon, Jong-Woo;Kim, Yong-Kee;Lee, Hoi-Young;Lee, Hyang-Woo;Han, Jeung-Whan
    • Archives of Pharmacal Research
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    • v.25 no.5
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    • pp.685-690
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    • 2002
  • The mitogen-stimulated serine/threonine kinase $p70^{S6k}$ plays an important role in the progression of cells from $G_0/G$_1$$ to S phase of the cell cycle by translational up-regulation of a family of mRNA transcripts family of mRNA transcripts which contain polypyrimidine tract at their 5 transcriptional start site. Here, we report that $p70^{S6k}$ was constitutively phosphorylated and activated to various degrees in serum-deprived AGS, A2058, HT-1376, MG63, MCF7, MDA-MB-435S, MDA-MB-231 and MB-157. Rapamycin treatment induced a significant dephosphorylation and inactivation of $p70^{S6k}$ in all cancer cell lines, while wortmannin, a specific inhibitor of PI3-K, caused a mild dephosphorylation of $p70^{S6k}$ in AGS, MDA-MB-435S and MB-157. In addition, SQ20006, methylxanthine phosphodiesterase inhibitor, reduced the phosphorylation of $p70^{S6k}$ in all cancer cells tested. Consistent with inhibitory effect of rapamycin on $p70^{S6k}$ activity, rapamycin inhibited [$^3H$]-thymidine incorporation and increased the number of cells at $G_{0}G_{1}$ phase. Furthermore, these inhibitory effects were accompanied by the decrease in growth of cancer cells. Taken together, the results indicate that the antiproliferative activity of rapamycin might be attributed to cell cycle arrest at $G_{0}G_{1}$ phase in human cancer cells through the inhibition of constitutively activated $p70^{S6k}$ of cancer cells and suggest $p70^{S6k}$ as a potential target for therapeutic strategies aimed at preventing or inhibiting tumor growth.