• Title/Summary/Keyword: Enzymatic pattern

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Preparation of $Gal^3Man_4(6^3-mono-{\alpha}-D-galacto-pyranosyl-{\beta}-mannotetraose)$ by Bacillus sp. ${\beta}-mannanase$ and Growth Activity to Intestinal Bacteria (Bacillus sp.유래 ${\beta}-mannanase$에 의한 $Gal^3Man_4(6^3-mono-{\alpha}-D-galacto-pyranosyl-{\beta}-mannotetraose)$ 조제 및 장내세균에 대한 생육활성)

  • Kim, Sang-Woo;Park, Gwi-Gun
    • Applied Biological Chemistry
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    • v.47 no.4
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    • pp.379-383
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    • 2004
  • For the elucidation of substrate specificity to the brown copra meal by Bacillus sp. ${\beta}-mannanase.$, the enzymatic hydrolysate after 24 hr of reaction was heated in a boiling water bath for 10 min, and then centrifuged to remove the insoluble materials from hydrolysates. The major hydrolysates composed of D.P 5 and 7 galactosyl mannooligosaccharides. For the separate of galactosyl mannooligosaccharides, the supernatant solution of 150 ml was put on a first activated carbon column. The column was then washed with 5 l of water to remove mannose and salts. The oligosaccharides in the column were eluted by a liner gradient of $0{\sim}30%$ ethanol, at the flow rate of 250 ml per hour. The sugar composition in each fraction tubes was examined by TLC and FACE analysis. The combined fraction from F3 was concentrated to 30 ml by vacuum evaporator. Then put on a second activated carbon column. The oligosaccharides in the column were eluted by a liner gradient of $0{\sim}30%$ ethanol (total volume: 5 l), at the flow rate of 250 ml per hour. The eluent was collected in 8 ml fraction tubes, and the total sugar concentration was measured by method of phenol-sulfuric acid. The major component of F2 separated by 2nd activated carbon column chromatography were identified $Gal^3Man_4(6^3-mono-{\alpha}-D-galactopyranosyl-{\beta}-mannotetraose)$. To investigate the effects of brown copra meal galactomannooligosaccharides on growth of Bifidobacterium longum, B. bifidum were cultivated individually on the modified-MRS medium containing carbon source such as $Gal^3Man_4$, compared to those of standard MRS medium.

Enzymological Properties of the Alkaline AL-Protease from Arthrobacter luteus and Detection of Its Active Amino Acid Residue (Arthrobacter luteus로부터 유래한 염기성 AL-Protease의 효소학적 성질 및 활성 아미노산 잔기의 검색)

  • Oh, Hong-Rock;Aizono, Yasuo;Funatsu, Masaru
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.13 no.2
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    • pp.193-204
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    • 1984
  • The enzymatic properties of the alkaline AL-protease, which had been prepared from the crude zymolyase of Arthrobzoter luteus, was investigated together with its active amino acid residue. Complete inactivaton of the proteolytic activity of AL-protease by either DFP or PMSF was simultaneously accompanied by the loss of its lytic effect on the lysis of yeast cell wall. In the reaction, AL-protease showed the pattern of inactivation to decrease very slowly, as compared to that of chymotrypsin, and that enzyme and DFP were found to react with a molar ratio of 1 : 1. The preparation of AL-protease exhibited no hydrolytic activity in any substrates of polysaccharases, playing a significant role in the lysis of yeast cell wall. The optimum pH and temperature of AL-protease was pH 10.5 and $65^{\circ}C$, respectively. It also showed stability in the pH range from 5 to 11 and at the temperature below $65^{\circ}C$. Through the identification of the amino acid residue in the active site of the $^{32}P$-diisopropylph-osphorylated(DIP) AL-protease modified specifically with $^{32}P$-labeled DFP, AL-protease was found to be a DFP-sensitive which has a mole of active serine residue involved in its proteolytic activity per mole of the enzyme.

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Acetylcholinesterase Inhibitory Effect of Green Tea Extracts According to Storage Condition (저장 조건에 따른 녹차 추출물의 acetylcholinesterase(AChE) 저해 효과)

  • Kwak, Ji Hyun;Jeong, Chang-Ho;Kim, Ji Hye;Choi, Gwi Nam;Shin, Young-Hee;Lee, Seung-Cheol;Cho, Sung Hwan;Choi, Sung-Gil;Heo, Ho Jin
    • Korean Journal of Food Science and Technology
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    • v.41 no.4
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    • pp.435-440
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    • 2009
  • Acetylcholinesterase (AChE) inhibitors, which enhance cholinergic transmission by reducing the enzymatic degradation of acetylcholine, are the only source of the compound that is currently approved for the treatment of Alzheimer's disease (AD). In these experiments, the concentration samples of green tea extracts were 100, 500, and 1,000 ${\mu}g$/mL. Among them, the highest concentration (1,000 ${\mu}g$/mL) of fresh green tea extracts showed the most potent inhibitory effect on AChE by reducing more than 40% of enzyme activity, and in a concentration-dependent pattern. In addition, we examined the effect of other storing conditions on AChE inhibition. In order to maintain storage for 3 months, the materials were held at the certain storing conditions of temperature (room temperature, 4 and $-20^{\circ}C$) and for water activity (0.81, 0.69, and 0.23). In these storing conditions, the difference in temperature did not contribute to the AChE inhibitory effect. Our presented data showed that the AChE inhibitory effect was affected by the concentration of green tea extract and by water activity (0.81). These results suggest that green tea may serve as a potential dietary source of AChE inhibitor.

Expression Analysis of Programmed Cell Death Genes in Porcine Parthenogenesis (돼지 단위생식란의 세포사멸 유전자 발현 양상에 관한 연구)

  • Son, Jong-Yoon;Kim, Sang-Hwan;Jung, Duk-Won;Ryu, Chun-Yeol;Yoon, Jong-Taek
    • Journal of Embryo Transfer
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    • v.30 no.3
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    • pp.239-248
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    • 2015
  • The nature of molecular mechanisms governing embryonic cell block is largely unknown, but recent reports have demonstrated that proper execution of programmed cell death is crucial for this process. The main objective of this study is to determine effects of programmed cell death on porcine oocytes development in vitro after parthenogenesis. Among the blastocysts matured in 3MA, MAP1LC3A and ATG5 RNA gene expression level increased in the order of Cyst < 3MA < RP. However, Casp-3 and TNF-r RNA gene expression level decreased in the order of RP < 3MA < Cyst. Expression of mTOR within the RP-cultured blastocyst was the most highly to the inner cell mass, while 3MA-cultured blastocyst showed very lowest expression in inner cell mass. The expression of mTOR showed a pattern opposite to that of MAP1LC3A. That is, its expression was the lowest in Cyst group. When the enzymatic activity of MMP-2 and MMP-9 was assessed in culture, the level of active MMP-9 was higher expression in the medium of each RP treatment group, with the level of another treatment group being relatively higher. Analyses of TIMP-2 and TIMP-3 revealed that their expression was higher in groups that did not receive RP treatment. More specifically, the level of TIMP-2 was not affected by Cyst treatment, while the level of TIMP-3 was higher in 3MA and RP treatment group. There was highly cell division activation efficiency of parthenogenesis on cultured system of RP supplement IVC medium. Therefore, these results suggest that embryo development was significantly increased in conditional culture medium with active autophagy as compared to common cultured condition. Further investigation of this distinction may enable the development of innovative improvements for the production of porcine somatic cell nuclear transfer.

A comparative Study on the Activities of Glutamic Dehydrogenase and Glutamic Transaminase in Livers of the Crucian carp, Pigeon, and Rat (붕어, 비둘기, 흰쥐의 肝臟의 Glutamic Dehydrogenase 및 Glutamic Transaminase 의 活性에 관한 比較 硏究)

  • Kim, Yong Kyu;Nam, Sang Yul
    • The Korean Journal of Zoology
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    • v.12 no.2
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    • pp.50-56
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    • 1969
  • The present investigation involves a comparative study of enzymatic activities in various animals. The levels of the liver protein of rat (22.0$\pm$0.01mg/ml and pigeon (22.0$\pm$ 0.16) are twice as high as that of crucian carp (13.0$\pm$ 0.09)(p < 0.01). Generally, the specific acitivity (3.77 $\pm$0.18 unit/mg) of rat glutamic pyruvic transaminase (GPT) is highest, pigeon intermediate (1.93 $\pm$0.01), and crucian carp lowest (0.71$\pm$0.07). On the other hand, the specific activity (8.23$\pm$0.09 unit/mg)of rat glutamic oxaloacetic transaminase (GOT) is highest, pigeon intermediate (3.95$\pm$0.09), and crucian carp lowest (0.92$\pm$0.01) (p < 0.01). Ratios of GOT activity to GPT activity appear no remarkable difference from the levels of various animals. Specific activity of glutamic dehydrogenase(GDH) in pigeon tissue exceeds those of rat and crucian carp. In liver, rat GOT specific activity is greater than crucian carp and pigeon. On the other hand, pigeon GDH specific activity is greater than those of rat and crucian carp. This wouls seem to be in accord with protein metabolic intensity. The patterns for GDH isozyme were remarkably appeared in various animals. Glutamic dehydrogenase isozymes gave different electrophoretic mobilities in various animals. It is interesting that crucian carp, pigeon, and rat would show this difference, which may be indicative of an evolutional pattern. The fact that livers in various animals show quite different enzyme activities would suggest the existence of such a general phylogenetic relationship.

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The Predictors of Effectiveness on Urokinase Instillation Therapy into Loculated Pleural Effusion. (소방이 형성된 흉막질환에서 유로키나제 주입치료의 예후인자)

  • Song, Kee-San;Bang, Jei-So;Kwak, Seung-Min;Cho, Chul-Ho;Park, Chan-Sup
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.3
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    • pp.621-628
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    • 1997
  • Background : As the pleural inflammation progresses, exudative pleural fluid becomes loculated rapidly with pleural thickening. Complete drainage is important to prevent pleural fibrosis, entrapment and depression of lung function. Intrapleural urokinase instillation therapy has been advocated as a method to facilitate drainage of gelatinous pleural fluid and to allow enzymatic debriment of pleural surface. This study was designed to investigate the predictors of effectiveness of intrapleural urokinase in the treatment of loculated pleural effusion. Method : Thirty-five patients received a single radiographically guided pig-tail catheter ranging in size from 10 to 12 French. Twenty-two patients had tuberculous pleural effusions, and 13 had non-tuberculous postpneumonic empyemas. A total of 240,000 units of urokinase was dissolved in 240 ml of normal saline and the aliquots of 80mL was instilled into the pleural cavity via pig-tail catheter per every 8hr. Effectiveness of intrapleural urokinase instillation therapy was assessed by biochemical markers, ultrasonography, and technical details. A greater than 50% improvement on follow-up chest radiographs was defined as success group. Result : Twenty-seven of 35 (77.1%) patients had successful outcome to urokinase instillation therapy. Duration of symptoms before admission was shorter in success group ($11.8{\pm}6.9day$) than in failure group ($26.62{\pm}16.5day$) (P<0.05). Amount of drained fluid during urokinase therapy was larger in success group ($917.1{\pm}392.7ml$) than in failure group ($613.8{\pm}259.7ml$) (P<0.05). Pleural fluid glucose was higher in success group ($89.7{\pm}35.9mg/dl$) than in failure group ($41.2{\pm}47.1mg/dl$) (P<0.05). Pleural fluid LDH was lower in success group ($878.4{\pm}654.3IU/L$) than in failure group ($2711.1{\pm}973.1IU/L$) (P<0.05). Honeycomb septated pattern on chest ultrasonography was observed in six of eight failure group, but none of success group (P<0.05). Conclusion : Longer duration of symptoms before admission, smaller amount of drained fluid during urokinase therapy, lower glucose value, higher LDH value in pleural fluid examination, and honeycomb septation pattern on chest ultrasonograph were predictors for failure group of intrapleural urokinase instillation therapy.

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On-site Output Survey and Feed Value Evaluation on Agro- industrial By-products (농산업부산물들에 대한 배출 현장 조사 및 사료적 가치 평가)

  • Kwak, W. S.;Yoon, J. S.
    • Journal of Animal Science and Technology
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    • v.45 no.2
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    • pp.251-264
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    • 2003
  • This study was conducted to make on-site survey on the output pattern and utilization situation of 19 by-products selected, to evaluate their nutritional characteristics, to find out a reliable index with which digestion of by-products can be predicted on the basis of chemical compositions analyzed and to diagnose the risk of using book values in the absence of the actual values analyzed for diet formulation. Production and utilization situations of by-products were quite various. Nutritionally, fruit processing by-products such as apple pomace (AP), pear pomace (PP), grape pomace (GP), and persimmon peel (PSP), and bakery by-products (BB) were classified as energy feeds. Soybean curd meal (SCM), animal by- products such as blood (BD), feather meal (FM) and poultry by-products (PB), and activated milk processing sludge (AMS) were classified as protein feeds. Soy hulls (SH), spent mushroom compost (SMC), barley malt hulls (BMH), waste paper (WP) and broiler litter (BL) were classified as roughage. Rumen contents (RC) and restaurant food waste (FW) were nutritionally analogous to complete diets for cattle and swine, respectively. Compared to soybean meal (SBM), BD and FM contained high (P<0.05) levels of amino acids and barley malt sprouts (BMS), AMS and FW contained low (P<0.05) levels of amino acids. Enzymatic (pepsin) digestibilities of proteinaceous feeds ranged between 99 and 66%. In vitro DM digestibility was high (P<0.05) in the order of FW, BB, AP, SH, PP, PSP, BMH, BMS, SCM, GP, RC, PB, BL, WP, SMC, AMS, FM and BD. In vitro DM digestibility had the highest correlation (r=0.68) with nonfibrous carbohydrate among chemical components. Differences between analyzed values of chemical components and book values were considerable. Caution is required in using book values when large amount of by-products are used in diets.