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Biochemical Characterization of a Novel Thermostable Esterase from the Metagenome of Dokdo Islets Marine Sediment (독도 심해토 메타게놈 유래 신규 내열성 에스테라아제의 생화학적 특성규명)

  • Lee, Chang-Muk;Seo, Sohyeon;Kim, Su-Yeon;Song, Jaeeun;Sim, Joon-Soo;Hahn, Bum-Soo;Kim, Dong-Hern;Yoon, Sang-Hong
    • Microbiology and Biotechnology Letters
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    • v.45 no.1
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    • pp.63-70
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    • 2017
  • A functional screen of 60,672 fosmid metagenomic clones amplified from marine sediment obtained from the Dokdo islets in Korea identified the gene EstES1, whose product, EstES1, displayed lipolytic properties on tributyrin-supplemented media. EstES1 is a 576 amino acid protein with a predicted molecular weight of 59.4 kDa including 37 N-terminal leader amino acids. EstES1 exhibited the highest sequence similarity (44%) to a carboxylesterase found in Haliangium ochraceum DSM14365. Phylogenetic analysis indicated that EstES1 belongs to a currently uncharacterized family of lipases. Within the conserved domain, EstES1 retains the catalytic triad that consists of the consensus penta-peptide motif, GESAG. EstES1 demonstrated a broad substrate specificity toward the long acyl group of ethyl esters (C2-C12), and its optimal activity was recorded toward p-Nitrophenyl butyrate (C4) at pH 9.0 and $40^{\circ}C$ (specific activity of 255.4 U/mg). The enzyme remained stable in the ranges of $60-65^{\circ}C$ and pH 9.0-10.5 and in the presence of methanol, ethanol, isopropanol, and dimethyl sulfoxide. Therefore, EstES1 has potential for use in industrial applications involving high temperature, organic solvents, and/or alkaline conditions.

The Proteinase Distributed in the Intestinal Organs of Fish 2. Characterization of the Three Alkaline Proteinases from the Pyloric Caeca of Mackerel, Scomber japonicus (어류의 장기조직에 분포하는 단백질분해효소에 관한 연구 2. 고등어 유문수조직중에 분포하는 3종 알칼리성 단백질분해효소의 특성)

  • KIM Hyeung-Rak;PYEUN Jae-Hyeung
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.19 no.6
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    • pp.547-557
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    • 1986
  • The characteristics of the three alkaline proteinases, Enz. A, B and C, from the pyloric caeca of mackerel have been investigated. The optimum condition for the activity of the Enz. A, B and C was pH 9.4, 9.8 and 9.8 at $45^{\circ}C$ for $2\%$ casein solution, and was pH 9.2 10.2 and 9.8 at $45^{\circ}C$ for $5\%$ hemoglobin denatured by urea, respectively. Enz. A, B and C by heat treatment at $50^{\circ}C$ for 5 min were inactivated 90, 33 and $37\%$, respectively, over the original activity. The reaction rate of the three alkaline proteinases was constant to the reaction time to 40 min in the reaction condition of $2{\mu}g/ml$ of enzyme concentration and $2\%$ casein solution. The reaction rate equation and Km value against casein substrate determined by the method of Lineweaver and Burk were: Enz. A, Y=3.6X and $Km=5.0{\times}10^{-3}\%$; Enz. B, Y=6.0X and $Km=1.0{\times}10^{-3}\%$; Enz. C, Y=4.2X and $Km=3.6{\times}10^{-3}\%$. The three alkaline proteinases were inactivated by $Ag^+$ and $Hg^{2+}$, but activated by $Mn^{2+},\;Sn^{2+}\;and\;Pb^{2+}$, Enz. B and C were remarkably inhibited by the soybean trypsin inhibitor. Molecular weight of Enz. A, B and C determined by SDS-polyacrylamide gel electrophoresis and Sephadex G-100 gel filtration was in the range of $27,500{\pm}2,500,\;20,500{\pm}1,500\;and\;15,250{\pm}250$, respectively.

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The Effects of Vitamin C on Lipid Contents and Fatty Acid Compositions of Serum and Liver in Rats Treated with Radiation or Aflatoxin B1 (Vitamin C가 방사선과 Aflatoxin B1을 투여한 흰쥐의 혈청과 간장의 지질성분 및 지방산 조성에 미치는 영향)

  • Kang, Jin-Soon;Kim, So-Young;Kim, Hee-Suk;Cho, Heung-Lae;Chai, Gyu-Young;Chung, Duck-Hwa
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.2
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    • pp.163-173
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    • 2007
  • Lipid peroxidation is one of the main manifestations of oxidative damage and has been found to play an important role in the toxicity and carcinogenesis of many carcinogens. This study was carried out to determine the effects of vitamin C on lipid contents and fatty acid compositions of serum and liver in male rats treated with radiation or aflatoxin $B_1\;(AFB_1)$. Six week-old male Sprague-Dawley rats were randomly divided into 7 groups; control group, radiation exposed group, $AFB_1$ treated group, X-ray and $AFB_1$ co-treated group. Three groups, except control group, were each further divided into vitamin C administered group and not administered groups. For this study, vitamin C was injected with 10 mg/kg of body weight by intraperitoneal injection and 1 hr later, 0.4 mg/kg of $AFB_1$ was injected by the same method. These administrations were repeated every 3 days over a period of 15 days. Only one time, X-ray was irradiated on whole liver with 1,500 cGy. Then vitamin C and AFB1 were administered by the same level and same method described above. On the 16th day of treatments, the animals were sacrificed. From the analysis of the serum lipid patterns, significant decrease (p<0.01) in triglyceride (TG) and total cholesterol levels were observed in X-ray and $AFB_1$ co treated group administered with vitamin C (group 7). In liver lipids, the levels of free cholesterol and total cholesterol were also decreased in X-ray and $AFB_1$ co treated group administered with vitamin C (group 7). The levels of serum free cholesterol and hepatic TG were not significantly different among all groups according to vitamin C administrations. The high density lipoprotein (HDL)-cholesterol level of serum was significantly (p<0.01) increased while the low density lipoprotein (LDL)-cholesterol level was decreased in X-ray and $AFB_1$ co treated group administered with vitamin C (group 7). In the phospholipid fatty-acid compositions of serum and liver tissue, group 3, 5 and 7 showed an increase in polyunsaturated fatty-acid (PUFA) but a decrease in saturated fatty acid (SFA) when compared to the control group. The composition ratio of fatty acid varied according to vitamin C administration. These results suggested that vitamin C has partly suppressive effects on lipid contents and fatty acid composition of serum and liver in rats treated by radiation and $AFB_1$.

A Study on the Antidotal Effects of Dietary Garlic Juice on Lead Poisoning Rats (납중독 흰쥐에서 식이 마늘 즙의 해독효과에 관한 연구)

  • Sheo, Hwa-Jung;Seo, Yu-Seok
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.3
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    • pp.342-350
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    • 2005
  • In the present work antidotal effect of dietary garlic was studied on lead-intoxicated rat. One of 5 groups of young Wistar sp. male rat, aged 4 weeks for control were fed only normal diet. Lead (25 ㎎/㎏.bw/week) was administered to other four groups for plumbism model over 4 weeks, of which three groups were supplemented with one of the following raw garlic juice: 1.10 (1% diet), 2.21 (2%) and 3.31 (3%) ㎎/㎏.bw/day respectively. Body weight gain rates in all garlic group significantly increased, especially in 2% garlic group that showed 9.8% net gain, as compared with only-lead treated group but lower values than control. The fecal and urinary lead excretion in all garlic groups significantly increased in a dose dependent fashion with highest value of 9.59% net gain in 3% garlic group as compared to lead treated control group. In comparison with lead treated control group, all garlic groups showed significantly increased hemoglobin contents, hematocrit values (Hct), red blood cell (RBC) count, mean corpuscular volume (MCV), and δ-amino levulinic acid dehydratase (δ-ALAD) activities. The values of 2% and 3% garlic groups remarkably increased while no significant difference between the values of 2% and 3% garlic groups was observed. The ALT activities, blood urea nitrogen (BUN) and creatinine (CR) in all garlic groups significantly decreased as compared with lead-treated control group. The values of 2% garlic group were the lowest and significantly different from the values of 1% and 3% garlic groups. The results showed that 2%-3% garlic juice in diet had obviously antidotal effects in lead-poisoned rats by promoting lead excretion. However, mega dose garlic such as in 3% garlic group might have some adverse effects on hepatic and renal functions in rats. In conclusion, the dietary habit to take ordinary garlic sauce in appropriate amount, may be helpful for preventing lead or other heavy metal intoxication.

Studies on the Processing of Krill Sauce (크릴간장 제조(製造)에 관한 연구(硏究))

  • Lee, Eung-Ho;Cho, Soon-Yeong;Cha, Yong-Jun;Park, Hyang-Suk;Kwon, Chil-Sung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.13 no.1
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    • pp.97-106
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    • 1984
  • The Antarctic krill, Euphausia superba, is drawing attention over the world as the largest source of unutilized proteins in the ocean. For the use of krill as a human food, processing conditions of krill sauce by autolysis and/or commercial proteolytic enzyme digestion were examined. The krill was chopped and mixed with equal weight of water, and hydrolyzed by autolysis and/or commercial proteolytic enzyme digestion. The optimal conditions for hydrolysis of krill were $52.5^{\circ}C$, pH 7.0-7.5, 3 hours by autolysis, $52.5^{\circ}C$, pH 6.3, 3hours by bromelain (0.5 %) digestion, and $52.5^{\circ}C$, pH 7.0-7.5, 3 hours by commercial complex enzyme (5 %) digestion, respectively The maximum hydrolyzing rate of protein were 83.2 % by autolysis, 89.7 % by bromelain digestion, 92.7 % by commercial complex enzyme digestion. After krill meat hydrolyzed by autolysis at optimum condition, inactivated at $100^{\circ}C$ for 20 minutes and filtered with Buchner funnel. Two kinds of products were prepared with krill hydrolysate and preservatives: one contained 10 % of sodium chloride and 0.06 % of benzoic acid and the other 10 % of sodium chloride and 3 % of ethyl alcohol. These products were filled in the sterilized glass bottle and sealed. The pH, volatile basic nitrogen, amino nitrogen, color value (L, a and b values) and viable counts of bacteria were determined during storage at $37^{\circ}C$. The results showed that the products could be preserved in good condition during one month at $37^{\circ}C$. As a method to reduce the sodium level in krill sauce, it is convinced that sodium chloride could be replaced half in partially by potassium chloride. In the products prepared from krill by autolysis, bromelain or commercial complex enzyme digestion, hypoxanthine and 5'-IMP were abundant among the nucleotides and their related compounds as 15.3-20.4 ${\mu}mole/g$, dry solid, 2.2-2.5 ${\mu}mole/g$, dry solid, respectively. The abundant free amino acids were lysine, leucine, proline, alanine and valine. The contents of these amino acids were 67.4 %, 69.4 %, 69.8 % of the total free amino acids of each products. And TMAO, betaine and total creatinine were low in contents. The flavor of krill sauce prepared from krill by autolysis or enzyme digestion was not inferior to that of traditional Kerean soy sauce by sensory evaluation.

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Anti-Inflammatory Effect of Ethanol Extract from Onion (Allium cepa L.) Peel on Lipopolysaccharide-Induced Inflammatory Responses in RAW 264.7 Cells and Mice Ears (LPS로 유도된 RAW 264.7 세포와 마우스 귀 조직에 대한 양파(Allium cepa L.) 껍질 에탄올 추출물의 항염증 효과)

  • Ahn, Na-Kyung;Kang, Bo-Kyeong;Kim, Koth-Bong-Woo-Ri;Kim, Min-Ji;Bae, Nan-Young;Park, Ji-Hye;Park, Sun-Hee;Ahn, Dong-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.11
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    • pp.1612-1620
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    • 2015
  • Inflammation is a complex process involving a variety of immune cells, which defend the body from harmful stimuli. However, pro-inflammatory cytokines and inflammatory mediators can also exacerbate diseases such as cancer. Onion peel contains several phenolic compounds, including quercetin at an amount 20 times greater in peel than edible flesh. Therefore, in this study, the anti-inflammatory effects of onion peel ethanol extract (OPEE) were investigated lipopolysaccharide-induced inflammatory response. In our results, NO production decreased in a dose-dependent manner. Secretion of IL-6, $TNF-{\alpha}$, and $IL-1{\beta}$ was suppressed by 44%, 53%, and 60% respectively, at $100{\mu}g/mL$. Moreover, OPEE also suppressed expression of COX-2, iNOS, $NF-{\kappa}B$, and MAPKs in a dose-dependent manner. Formation of mice ear edema was reduced at the highest dose tested compared to the control, and reduction of ear thickness was observed in the histological analysis as well. In the acute toxicity test, no morality was observed in mice administered 5,000 mg/kg body weight of OPEE over a 2-week observation period. These results suggest that OPEE may have significant effects on inflammatory factors and be a potential anti-inflammatory material.

The Result of Bone Grafting for Fibrous Dysplasia (섬유성 골 이형성증에서 시행한 골 이식술의 결과 분석)

  • Jeong, Won-Ju;Kim, Tae-Seong;Cho, Hwan-Seong;Yoon, Jong-Pil;Park, Il-Hyung
    • The Journal of the Korean bone and joint tumor society
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    • v.20 no.2
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    • pp.74-79
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    • 2014
  • Purpose: Fibrous dysplasia is related to the mutation of gene encoding the alpha-subunit of a signal-transducing G-protein and has variable clinical course. Operation can be performed to prevent functional disorder or structural deformity. After curettage, autologous bone graft were used to fill the defects after curettage. The aim of this study is to compare the result of autogenous cancellous bone grafting and allogenic bone grafting for fibrous dysplasia. Materials and Methods: Among the patients who visit our hospital during the period of April, 1997 to October, 2013, we selected 34 patients who diagnosed fibrous dysplasia and visited our clinic over 1 year. There were 13 males and 21 females. Average age was 26.4 (range 2 to 57) years old. Autogenous bone graft (group I) in 5 cases, Non-autogenous bone graft (group II) in 30 cases. Iliac bone is used in all cases of autogenous bone graft. There were no significant difference in age, follow-up period, preoperational laboratory finding between two groups. Radiographic image was done to evaluate the recurrence of fibrous dysplasia or secondary degeneration. Results: There were four cases in recurrence (group I: 1 case, group II: 3 cases, p=0.554). In all recurrent cases, reoperations were done using curettage and autogenous iliac bone graft. There was no re-recurrence after reoperation. One case of secondary aneurysmal bone cyst was confirmed (group II) and 1 cases of pathologic fractures had developed (group I: 0 case, group II: 1 cases, p=0.559). No malignant change occurred. Conclusion: There were no significant difference between autogenous bone graft group and non-autogenous bone graft group. Our result suggested that autogenous bone graft seems to be good method to treat fibrous dysplasia, in the case of small volume of tumor lesion or non-weight bearing portion.

Anti-inflammatory Effect of Ethanol Extract from Sargassum fulvellum on Lipopolysaccharide Induced Inflammatory Responses in RAW 264.7 Cells and Mice Ears (LPS로 유도된 RAW 264.7 세포와 마우스 귀조직에 대한 참모자반 (Sargassum fulvellum) 에탄올 추출물의 항염증 효과)

  • Jeong, Da-Hyun;Kim, Koth-Bong-Woo-Ri;Kim, Min-Ji;Kang, Bo-Kyeong;Bark, Si-Woo;Pak, Won-Min;Kim, Bo-Ram;Ahn, Na-Kyung;Choi, Yeon-Uk;Ahn, Dong-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.8
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    • pp.1158-1165
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    • 2014
  • This study investigated the anti-inflammatory effects of Sargassum fulvellum ethanol extract (SFEE) on the lipopolysaccharide (LPS)-induced inflammatory response. SFEE remarkably suppressed production of NO and pro-inflammatory cytokines (IL-6, $TNF-{\alpha}$, and $IL-1{\beta}$ at 50 and $100{\mu}g/mL$. There were no cytotoxic effects on proliferation of macrophages treated with SFEE compared to the control. SFEE reduced expression of iNOS and COX-2 proteins in a dose-dependent manner. The formation of edema in mouse ears was reduced at the highest dose tested compared to the control. Moreover, in the acute toxicity test, no mortality occurred in mice administered 5,000 mg/kg body weight of SFEE over the 2-week observation period. These results suggest that SFEE may have significant effects on inflammatory factors and be a potential anti-inflammatory therapeutic material.

A Study on Nutrient Intake Status and Dietary Behaviors in Elementary School Children by Body Shape (체형에 따른 일부 초등학교 아동의 영양소 섭취상태 및 식행동에 관한 조사)

  • Han, Gi-Hyung;Kim, Gyeong-Eup;Kim, Sung-Hee
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.10
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    • pp.1385-1393
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    • 2006
  • The purpose of this study was to investigate nutrient intake status and dietary behaviors in elementary school children. The subjects were 98 boys and girls in the 4~6 grade and divided into three groups by body shape. The protein intakes of the subjects were higher than the Korean EER (estimated energy requirements). Especially male over weight group had more than twice intakes (216.0% ) comparing the Korean EER. The mean daily intakes of calcium, iron, vitamin A and vitamin C in all subjects did not meet the Korean RI (recommended intake). The vitamin C intake of male underweight group was only 43.2% comparing the Korean RI. Rice & cereals were main food sources of energy, protein and iron, and milk & milk products were main food sources of calcium. As their weights were high, the meal volume and the rate of skipped meal were shown to be high (p<0.05). Thus, this study implicates that desirable nutrition education may be needed for overweight and underweigh children.

Growth Performances of Seedlings of Daphniphyllum macropodum in Naejang National Park for Container Seeding Production (시설양묘에 적용을 위한 내장산 굴거리 나무 군락내 치수의 생장 특성)

  • Chung, Jin-Chul;Jeong, Ji-Young;Choi, Jeong-Ho;Jeon, Kyung-Soo;Bae, Jong-Hyang
    • Journal of Bio-Environment Control
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    • v.20 no.4
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    • pp.365-372
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    • 2011
  • This study aimed to examine foundational data for container seedling production with the subject of Daphniphyllum macropodum community in Mt. Naejang National Park. To achieve the goal, it investigated the growth characteristics of young tree seedlings growing in places with different light intensity environment. Regarding the growth environment of Daphniphyllum macropodum community, it was typical heavy rain summer climate, and the soil was silt loam with the organic content as 11.42~15.61%, total nitrogen as 0.50~0.76%, cation exchangeable capacity (C.E.C) as 18.92~23.32 cmol/kg, and pH as 4.85~5.58. About light intensity environment changed by research plots, relative transmittance of solar radiation was 71~76% in plot A, 37~42% in plot B, 65~70% in plot C, and 28~33% in plot D. The seedlings tended to be intensively distributed either under the crown of their mother tree or in the slope site, and plot A and C where light intensity environment is relatively more favorable showed 1,550 tree/ha and 1,250 tree/ha. Total biomass production of Daphniphyllum macropodum seedlings was 5.37 g in plot A and 5.29 g in plot C, so they were higher than 4.42~4.51 g in plot B and D with relatively less favorable light intensity environment. The T/R ratio was 1~2, leaf area rate was $139.71{\sim}183.50cm^2{\cdot}g^{-1}$, leaf area ratio was $39.68{\sim}60.66cm^2{\cdot}g^{-1}$, and leaf dry weight ratio grew higher in the range of $0.28{\sim}0.33cm^2{\cdot}g^{-1}$ as the intensity of radiation became less. It is thought that in the generation and growth of Daphniphyllum macropodum seedlings, the intensity of light has more effects than the organic content in soil. And it is also thought that in the application of container seedlings production, light environment management over 65~70% to full sun light intensity will affect significantly the initial growth of Daphniphyllum macropodum.