• Title/Summary/Keyword: Enzymatic activity

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Characterization of Prophange Cured Strain Derivative from Lactobacillus casei YIT 9018 (Lcatobacillus casei YIT 9018로 부터 분리한 Prophage Cured Strain의 특성)

  • Lee, Jeong-Jun;Oh, Tae-Kwang;Chang, Hyo-Ihl;Baek, Young-Jin
    • Microbiology and Biotechnology Letters
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    • v.22 no.5
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    • pp.467-476
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    • 1994
  • Lactobacillus casei HY 2782, prophage cured strain was characterized to be stable as much as L casei YIT 9018, parent strain. By southern hybridization, it was confirmed that the temperate phage was incorporated in chromosomal DNA of L. casei YIT 9018 as a prophage. It was also proved that the prophage was cured from chromosomal DNA of L casei HY 2782. The growth rate, lactic acid producing ability, carbohydrates fermentation, and enzymatic activity of L. casei HY 2782 were found to be similar to those of L. casei YIT 9018. When L casei HY 2782 was used as a host, the multiplicity of infection (M.O.I.) of the temperate phage for L. casei HY 2782 was 1.0~5.0. Restriction enzyme analysis of pLC90 plasmid from L. casei HY 2782 was shown that the size was an approximately 68.22 kb. The plasmid profiles, genomic DNA patterns, and cellular fatty acids composition of L. casei HY 2782 were similar to those of L casei YIT 9018. And the major fatty acids composition of these strains were C$_{14;0}$,C$_{16;1}$, C$_{16;0}$, C$_{18;1}$ and C$_{19;cyclo-}$ 10 sets of arbitrary primer in the PCR were screened to find differentiation against two strains of L. casei. Among them, b$_{5}-1/17-1 primer was produced an approximately 1.3 kb DNA band of only L casei YIT 9018. And b$_{5}-2/17-2 primer was produced an approximately 1.0 kb DNA band of only L casei HY 2782.

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Biological Constituents of Aged Garlic Extract as Biomarker (숙성마늘 extract 의 biomarker로서 생리활성 성분)

  • Yang, Seung-Taek
    • Journal of Life Science
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    • v.19 no.1
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    • pp.138-146
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    • 2009
  • Garlic (Allium sativum) are an agronomically important genus because of their sulfur flavour components. The majority of the volatiles flavour principles are generated through the enzymatic hydrolysis of the non-volatile organosulfur compounds. However, these compounds may be possible sources of new novel bioacuve and therapeutic principles. Garlic has strong antioxidant activity, and epidemiological studies support the fad that diets rich of garlic may prevent some of the chronic diseases. The health cares of garlic likely arise from a wide variety of components, which may work synergistically. The chemical changes of garlic composition makes it plausible that a variation in processing can lead to acquisition of differential chemical compositions of garlic products. Especially highly unstable allicin can easily disappear during processing and are quickly transformed into a various organosulfur compounds. Various supplements of garlic, particularly aged garlic extract (AGE), are known to possess a promising antioxidant potential and are effective in prevention of chronic diseases because of the bioactive constituents. Although all of active ingredients of AGE are not elucidated, water-soluble components of AGE, including S-allylcysteine, S-allylmercaptane, steroid saponins, tetrahydro-${\beta}$-carboline derivatives, and fructosyl-arginine, appears to be associated with the pharmacological effects of AGE. Consequently, the allicin free garlic components such as S-allylcysteine, S-allylmercaptane, steroid saponins, tetrahydro-${\beta}$-carboline derivatives, and fructosyl-arginine can be applicable to standardization of the quality of commercial garlic products. This review provides an insight into garlic's biomarkers and presents evidence that they may either prevent or delay chronic disease associated with aging.

Changes of Polyphenols and Polyphenol Oxidase Active Bands during Apple Wine Fermentation (사과주(酒) 발효과정중(醱酵過程中) Polyphenol성(性) 물질(物質) 및 Polyphenol Oxidase 활성(活性) Band의 변화(變化))

  • Chung, Ki-Taek;Seo, Seung-Kyo;Song, Hyung-Ik
    • Korean Journal of Food Science and Technology
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    • v.16 no.4
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    • pp.413-417
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    • 1984
  • To examine enzymatic browning of apple wine, changes of active bands of polyphenol oxidase (EC 1.10.3.1) as well as polyphenol substances related to browning of apple wine were investigated during wine brewing. The decrease of total phenol was remarkably inhibited by the addition of sodium metabisulfite. In the meantime, auto-oxidation of catechol in a model system increased proportionally as the reaction pH and temperature increased. Catechol oxidation, however, was not detected at $4^{\circ}C$ below pH 5.0. Polyacrylamidegel electrophoretic patterns showed that the apple (Jonathan) indicated 4 bands with polyphenol oxidase activity, designated a, b, c and d whose Rm were 0.21, 0.30, 0.41 and 0.51, respectively. Among these, 2 bands, a and c remained until 5th day fermentation and only c band after 6th day fermentation. After pasteurization of apple wine at $60^{\circ}C$ for 30min, c band also remained.

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Effects of Sagungsan on the Hemostasis, Intracranial Pressure, Blood Pressure and Cardiovascular System in Expreimental Animals (사궁산(莎芎散)이 실험동물(實驗動物)의 지혈(止血), 뇌압(腦壓), 혈압(血壓) 및 심혈관계(心血管系)에 미치는 영향(影響))

  • An, Il-Hoe;Park, Young-Soon;Kim, Seh-Gil
    • The Journal of Internal Korean Medicine
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    • v.15 no.1
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    • pp.80-98
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    • 1994
  • According to the original documents, Sagungsan is considered as an effective drug for controlling the hypertensive epistaxis induced by tension of autonomic nerve and it's hyperfunction. The present experiment was designed to understand the effect of Sagungsan extract on the hemostatic action, intracranial pressure, blood pressure and cardiovascular system in experimental animals. And thus the bleeding time, prothrombin time, capillary dilation, blood pressure, Intracranial pressure, and enzymatic analysis of the ATPase activities were studied. The result obtained here were as followings: 1. Sagungsan water extract reduced the bleeding time in mouse, and prolonged the prothrombin time in rabbits. 2. The drug extract increased the tail volume by capillary dilation in rats. 3. The drug extract inhibited the increase of intracranial pressure and arterial blood pressure in rabbits. 4. At the early time, the increase of arterial blood pressure by the drug extract significantly inhibited by pretreated atropin and regitine in rabbits. 5. The drug extract relaxed the smooth muscle by stimulating the Mg2+-Ca2+-ATPase activities of gastric sarcoplasmic reticulum isolated from rabbit stomach. 6. The drug extract stimulated the heart contraction by inhibiting the $Mg^{2+}-Ca^{2+}-ATPase$ activities of cardiac sarcoplasmic reticulum isolated from rabbit heart. The inhibitory mechanism was reversible and noncompatitive. 7. The drug extract increased the hepatic blood volume by stimulating the hepatic total ATPase activities and hepatic metabolism. 8. The drug extract acted as a tranquilizer by inhibiting the neural Na+-K+-ATPase activity. According to the results, Sagungsan water extract dilated the capillaries, stimulated the heart beat, and thus increased the blood flow with decreasing the intracranial pressure and blood pressure. These effects stanches the epistaxis collectively.

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A Molecular Modeling Study of AAD16034

  • Cho, Hoon;Choi, Cheol-Hee;Yoo, Kyung-Ho;Cho, Seung-Joo
    • Molecular & Cellular Toxicology
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    • v.4 no.4
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    • pp.307-310
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    • 2008
  • AAD16034 is an alginate lyase from Pseudoalteromonas sp. IAM14594. A very close homologue with known 3D structure exists (marine bacterium Pseudoalteromonas sp. strain no. 272). A three-dimensional structure of AAD16034 was generated based on this template (PDB code: 1J1T) by comparative modeling. The modeled enzyme exhibited a jelly-roll like structure very similar to its template structure. Both enzymes possess the characteristic alginate sequence YFKhG+Y-Q. Since AAD16034 displays enzymatic activity for poly-M alginate, docking of a tri-mannuronate into the modeled structure was performed. Two separate and adjacent binding sites were found. The ligand was accommodated inside each binding site. By considering both binding sites, a plausible binding pose for the poly-M alginate polymer could be deduced. From the modeled docking pose (i.e., the most important factor that attracts alginate polymer into this lyase) the most likely interaction was electrostatic. In accordance with a previous report, the hydroxyl group of Y345 was positioned close to the ${\alpha}$-hydrogen of ${\beta}$-mannuronate, which was suitable to initiate a ${\beta}$-elimination reaction. K347 was also very near to the carboxylatemoiety of the ligand, which might stabilize the dianion intermediate during the ${\beta}$-elimination reaction. This implies that the characteristic alginate sequence is absolutely crucial for the catalysis. These results may be exploited in the design of novel enzymes with desired properties.

(1) Studies on the Determination of the Rate Control Steps in the Various Metabolic Cycles (I) - The Affect of the Light to the Enzyme Activities in the Cotyledons of the Germinating Seeds - (대사과정(代謝過程)에 있어서 율속단계(律速段階) 결정(決定)에 관(關)한 연구(硏究) (I) - 발아종자자엽(發芽種子子葉)에 있어서 산소활성(酸素活性)에 미치는 광선(光線)의 영향(影響) -)

  • Sin, Gwi-Nam
    • Applied Biological Chemistry
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    • v.3
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    • pp.1-7
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    • 1962
  • In order to study the rate control step in the protein metabolic course of the chlorophyll formation, the transaminase activities which are obtained freely in the extracts of cotyledons of germinating peas at the light and the dark places, are measured in Beckman mopel D.U, spectrophoto meter at 490 mu. In this case, of two enzymatic reaction products; oxalacetic acid and pyruvic acid, the former is converted to pyruvic acid by aniline citrate and after each pyruvate phenyl hydrazones are extracted by toluenes: when this is treated with strong alcoholic alkali, a colored hydrazone is formed and it is measured by above apparatus. The estimated G.O.T. and G.P.T. in the germinated cotyledons at dark and light places considerably differ in their activities; G.O.T. and G.P.T. activities which are formed at the light are more increased than at the dark and also they differ in their rates through germination, though G.O.T. activity increment is smoothly but that of G.P.T. is more sharply, and they are considered to be directly affected to the chlorophyll formation and indirectly to the growth. G.O.T. and G.P.T. in each fractions of cell in the cotyledons should be formed by dissociation of zymogens in the microsomal fractions and it seems to promoted by light. In the formation of the chlorophyll, the protein metabolism occurred mainly in the microsomal fractions and the rate determining step is found at the point where the zymogene that is able to produce G.P.T. is activated, and this activation is promoted by light as noted above.

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Effects of acetaminophen administration on liver histopathology, serum GOT/GPT levels and circulating microRNA-122 concentration in olive flounder (Paralichthys olivaceus)

  • Najib, Abdellaoui;Kim, Dong Soo;Kim, Ki Hong
    • Journal of fish pathology
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    • v.29 no.1
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    • pp.25-33
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    • 2016
  • In human medicine, circulating microRNAs have been successfully utilized as early biomarkers for various abnormalities and disease states. Vertebrate miR-122 is a liver-specific microRNA which is expressed almost solely in hepatocytes and plays an important role in the regulation of hepatocyte function. In this study, to evaluate the potential utility of circulating miR-122 as a biomarker for liver injury in olive flounder (Paralichthys olivaceus), fish were orally intubated with two doses of acetaminophen (500 mg/kg or 1.000 mg/kg of body weight), and the expression of miR-122 in serum was quantified using real time-PCR. Histological change in liver, and the enzymatic activity of glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT) were also analyzed. The results showed that miR-122 was higher in acetaminophen administered groups compared to control group. The histopathological effect of acetaminophen on olive flounder liver was not distinct. The serum level of GPT and GOT was increased within 2 folds compared to control group by acetaminophen administration. However, the serum miR-122 level was increased more than 3 or 4 folds compared to the control group by administration of 1000 mg/kg of acetaminophen. These results suggest the possible use of miR-122 as an indicator of liver injury in olive flounder, even when histopathological effects are not distinctive.

Analysis of Mutant Tryptophan Synthases with Defective Enzymatic Properties (트립토판 합성효소 잔기 치환체의 효소성질 결함에 대한 분석)

  • Kim, Il;Shin, Hye-Ja;Kim, Han-Do;Im, Woon-Ki
    • Journal of Life Science
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    • v.14 no.2
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    • pp.252-254
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    • 2004
  • The mutant tryptophan synthases at $\alpha$Asp$^{56}$ have low catalytic activities. The enzymes were treated with $\alpha$ and $\beta$ subunit-specific ligands in the presence of L-serine and indoline. It was shown that the cations resulted in changes of absorbance patterns among the proteins, while glycerophosphate showed similar pattern of absorbance. The glycerophosphate binds to the active site of $\alpha$ subunit so that $\alpha$Asp$^{56}$ may not be involved in the allosteric control with the active site of $\alpha$ subunit occupied by substrate. The results suggest that $\alpha$Asp$^{56}$ may playa role in the step of a series of reaction occurring without bound substrates in the active site of asubunit.

Effects of Carthami flos on pacemaker potentials of small intestinal and colonic interstitial Cells of Cajal (홍화의 생쥐 소장 및 대장 카할 간질세포의 향도잡이 전위 조절에 미치는 효능에 관한 연구)

  • Kim, Byung Joo
    • Herbal Formula Science
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    • v.27 no.4
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    • pp.237-244
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    • 2019
  • Objectives : The purpose of this study was to investigate the effects of Carthami flos on pacemaker potentials of small intestinal and colonic Interstitial Cells of Cajal (ICC). Methods : To dissociate the ICC, we used enzymatic digestions from the small intestine and colon in mice. In the ICC, the electrophysiological whole-cell patch-clamp configuration was used to record pacemaker potentials in the cultured ICC. Results : 1. The ICC generated pacemaker potentials in the murine small intestine and colon. 2. Pretreatment with a Ca2+ free solution and thapsigargin, a Ca2+-ATPase inhibitor in the endoplasmic reticulum, stopped the pacemaker potentials. In the case of Ca2+-free solutions, Carthami flos did not induce membrane depolarizations in the murine small intestine and colon. However, when thapsigargin in a bath solution was applied, Carthami flos induced membrane depolarizations only in the murine colon. 3. Pretreatment with 2-APB (transient receptor potential melastatin (TRPM) channel inhibitor) abolished the pacemaker potentials and suppressed Carthami flos-induced effects in the murine small intestine and colon. 4. However, pretreatment with T16Ainh-AO1 (Ca2+ activated Cl- channel; anoctamin 1 (ANO1) inhibitor) did not affect the pacemaker potentials and induced Carthami flos-induced effects only in the murine small intestine. Conclusions : These results suggest that Carthami flos can modulate the pacemaker activity of ICC and the mechanisms underlying pacemaking in ICC might be different in the small intestine and the colon.

Expression of Bcl-2 in the epithelial lining and clinical findings of keratocystic odotogenic tumor (각화낭치성종양의 이장상피에서 Bcl-2 발현양상과 임상지표)

  • Lee, In-Hyuk;Choi, So-Young;Park, Ji-Hoon;Kim, Chin-Soo
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.37 no.3
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    • pp.161-168
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    • 2011
  • Purpose: The odontogenic keratocysts demonstrated a high recurrence rate and a biologically aggressive nature. This might be due to unknown factors inherent in the epithelium or enzymatic activity in the fibrous wall. Bcl-2 protein is characterized by its ability to inhibit apoptosis. The aim of this study was to evaluate the expression and distribution of bcl-2 in the OKCs, its possible relationship with the tumorous characteristics, such as the aggressive nature and high recurrence rate, and its usefulness to differentiate OKCs from dentigerous cysts. Materials and Methods: Formalin fixed paraffin-embedded tissue sections of 53 OKCs, and 44 dentigerous cyst were immunohistochemically analyzed quantitatively for the immunoreactivity of the bcl-2 protein with i-solution. Results: More Bcl-2 expression was observed in the OKCs (mean34.387%) than dentigerous cyst (mean11.144%) with statistical significance (P<0.001). Seventeen and 15 of the 32 OKCs in this study showed positivity in the basal layer and basal/suprabasal layers, respectively. In dentigerous cyst, 2 of 3 showed positivity in the basal cell layer. Conclusion: Considering that bcl-2 over expression may lead to the increased survival of epithelial cells, this study demonstrated a possible relationship between the aggressive nature of OKC and the intrinsic growth potential of its lining epithelium. Furthermore, the basal/suprabasal distribution of bcl-2 positive cells was observed in some OKCs, which might have a significant impact on the behavior of cysts. The bcl-2 expression of OKCs can be useful for differentiating OKCs from dentigerous cysts.