• Title/Summary/Keyword: enzyme activity change

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The Relationship between Structural Denaturation of Antioxidative Enzymes and Their Enzyme Activity due to Repeated Exposure to UV-A (UV-A 반복노출로 인한 항산화효소의 구조변성과 효소활성의 상관관계)

  • Park, Mijung;Yoo, Hyo Jung;Kim, Jong Chan;Kim, So Ra
    • Journal of Korean Ophthalmic Optics Society
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    • v.20 no.1
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    • pp.75-81
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    • 2015
  • Purpose: The present study was conducted to investigate whether the changes in structure and activity of antioxidative enzymes, superoxide dismutase(SOD) and catalase(CAT) present in the eyes appeared when they were repeatedly exposed to UV-A, and reveal the correlation of these changes. Methods: Each enzyme solution was prepared from the standardized SOD and CAT, and repeatedly exposed to UV-A of 365 min under the condition of 30 minutes, 1 hour and 2 hours a day over 1, 2, 3, 4 and 5 days. Structural denaturation of SOD and CAT induced by repeat UV-A irradiation was confirmed by the electrophoretic analysis, and their enzyme activity was determined by the colorimetric assay using the proper assay kit. Results: SOD exposed repeatedly to UV-A showed the polymerization pattern through the electrophoretic analysis when it was repeatedly exposed under the condition of at least 1 hour a day however, the change of its activity was found to be less than 12%. On the other hand, CAT repeatedly exposed to UV-A showed reduced size of the electrophoretic band which indicated a structure denaturation and its activity was significantly decreased. In the case of that the repeat exposure time was longer, CAT activity was completely lost even though some enzyme band was shown in the electrphoretic analysis. Conclusions: From these results, it was revealed that the degree and pattern in structural denaturation of antioxidative enzymes differently appeared according to the type of enzyme, and the degree of structural denaturation was not always consistent with the reduction in enzyme activity.

Effects of Pinus densiflora on soil chemical and microbial properties in Pb-contaminated forest soil

  • Kim, Sung-Hyun;Lee, In-Sook;Kang, Ho-Jeong
    • Journal of Ecology and Environment
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    • v.34 no.3
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    • pp.315-322
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    • 2011
  • We investigated the effect of Pb uptake by Pinus densiflora and the Pb fraction in forest soil. We also investigated the change in soil physicochemical characteristics, microbial activity, and root exudates of Pinus densiflora in Pb-contaminated soils. Three-year-old pine seedlings were exposed to 500 mg/kg Pb for 12 months. The metal fractions were measured using sequential extraction procedures. Additionally, factors that affect solubility (three soil enzyme activities and amino acids of root exudate compounds) were also determined. The results showed that Pb contamination significantly decreased enzyme activities due to soil characteristics. In addition, organic matter, nitrate content, and Pb concentration were time dependent. The results indicate that changes in the Pb fraction affected Pb uptake by pine trees due to an increase in the exchangeable Pb fraction. The concentrations of organic acids were higher in Pb-spiked soil than those in control soil. Higher rhizosphere concentrations of oxalic acid resulted in increased Pb uptake from the soil. These results suggest that pine trees can change soil properties using root exudates due to differences in the metal fraction.

Changes in Microflora and Enzyme Activities of Traditional Kochujang Meju during Fermentation (재래식 고추장 메주 숙성 중 미생물과 효소력의 변화)

  • Park, Jong-Myon;Oh, Hoon-Il
    • Korean Journal of Food Science and Technology
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    • v.27 no.1
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    • pp.56-62
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    • 1995
  • This study was designed to investigate the changes in microflora and enzyme activities of traditional kochujang meju during fermentation for 60 days. The pH of meju decreased continously up to 40 days of aging and then increased slightly thereafter, while the change in titratable acidity showed the opposite trend to that of pH. The viable cell count of aerobic bacteria increared gradually for up to 40 days of fermentation and then decreased slightly thereafter, while that of molds and yeasts showed a rapid increase up to 40 days of fermentation and then leveled off. ${\alpha}-amylase$ activity increased slightly for up to 40 days of meju fermentation and then stabilized. On the other hand, ${\beta}-amylase$ and glucoamylase activities did not show a significant change for up to 20 days of fermentation and then increased rapidly at 40th day of fermentation. Acidic, neutral and alkaline protease activities increased sharply up to 40 days of aging and then decreased significently at 60th day of fermentation. These results suggest that meju fermented for 40 days had the highest quality in terms of the number of microflora and enzyme activity.

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Overexpression of Recombinant Arylsulfatase Cloned from Pseudoalteromonas carrageenovora

  • Kim Jong-Oh;Kim Seok-Ryel;Lim Jae-Myung;Nam Soo-Wan;Kim Hyeung-Rak
    • Fisheries and Aquatic Sciences
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    • v.8 no.3
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    • pp.118-121
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    • 2005
  • Arylsulfatase cloned from a marine bacterium, Pseudoalteromonas carrageenovora, was over-expressed in Escherichia coli. Most of the recombinant arylsulfatase was found in the cell lysate with induction up to $10{\mu}M$ IPTG. However, enzyme activity was observed both in the culture supernatant and cell lysate by induction with IPTG concentration of $50-5,000{\mu}M$. Most of the recombinant enzyme was localized in the periplasmic space with $10{\mu}M$ IPTG induction, while half of the enzyme was distributed in the periplasmic space with $50{\mu}M$ IPTG induction. Cell growth and arylsulfatase activity did not change with the induction time, and the level of recombinant arylsulfatase expression was maintained at 4-5 U/mL after 6 to 14 hr of culture.

Enzyme-Catalyzed Henry Reaction in Choline Chloride-Based Deep Eutectic Solvents

  • Tian, Xuemei;Zhang, Suoqin;Zheng, Liangyu
    • Journal of Microbiology and Biotechnology
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    • v.26 no.1
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    • pp.80-88
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    • 2016
  • The enzyme-catalyzed Henry reaction was realized using deep eutectic solvents (DESs) as a reaction medium. The lipase from Aspergillus niger (lipase AS) showed excellent catalytic activity toward the substrates aromatic aldehydes and nitromethane in choline chloride:glycerol at a molar ratio of 1:2. Addition of 30 vol% water to DES further improved the lipase activity and inhibited DES-catalyzed transformation. A final yield of 92.2% for the lipase AS-catalyzed Henry reaction was achieved under optimized reaction conditions in only 4 h. In addition, the lipase AS activity was improved by approximately 3-fold in a DES-water mixture compared with that in pure water, which produced a final yield of only 33.4%. Structural studies with fluorescence spectroscopy showed that the established strong hydrogen bonds between DES and water may be the main driving force that affects the spatial conformation of the enzyme, leading to a change in lipase activity. The methodology was also extended to the aza-Henry reaction, which easily occurred in contrast to that in pure water. The enantioselectivity of both Henry and aza-Henry reactions was not found. However, the results are still remarkable, as we report the first use of DES as a reaction medium in a lipase-catalyzed Henry reaction.

Benzoyltransferase and Phenylacetyltransferase Activities in Cholestatic Rat Liver Induced by Common Bile Duct Ligation

  • Kim, Young-Jin;Kim, You-Hee
    • BMB Reports
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    • v.32 no.1
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    • pp.67-71
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    • 1999
  • We have investigated the effect of cholestasis on the closely related acyl-CoA:amino acid N-acyltransferase, benzoyltransferase, and phenylacetyltransferase activities in rat liver. Benzoyltransferase and phenylacetyltransferase activities in the liver cytosol, mitochondria, and microsome were investigated for a period of 42 d after common bile duct ligation. Both the mitochondrial and microsomal benzoyltransferases showed significant increase in their activities between the 1st and 7th day after common bile duct ligation, although the cytosolic benzoyltransferase activity did not show a significant change compared to the activities from the sham-operated control. The cytosolic phenylacetyltransferase activity showed a significant increase between the 1st and 2nd day, the mitochondrial activity showed a significant increase between the 2nd and 7th day, and microsomal activity showed a significant increase between the 1st and 7th day, respectively. Enzyme kinetic parameters of hepatic benzoyltransferase were analyzed using benzoyl coenzyme A as a substrate with the preparations from the 1st day post-ligation. Enzyme parameters of hepatic phenylacetyltransferase were also analyzed using phenylacetyl coenzyme A as a substrate with the preparations from the 2nd day post-ligation. The results indicated that although the $K_m$ values of these enzymes were about the same as the sham-operated control, the $V_{max}$ values of both enzymes increased significantly. These results, therefore, suggest that the biosynthesis of benzoyltransferase and phenylacetyltransferase has been induced in response to cholestasis.

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Hydrolysis of Egg Yolk Protein in a Packed Bed Reactor by Immobilized Enzyme (충진층 반응기에서 고정화 효소에 의한 난황 단백질의 가수분해)

  • Kang, Byung-Chul
    • Journal of Life Science
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    • v.20 no.11
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    • pp.1656-1661
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    • 2010
  • Alkaline protease for the hydrolysis of egg yolk protein was immobilized on five carriers - Duolite A568, Celite R640, Dowex-1, Dowex 50W and Silica gel R60. Duolite A568 showed a maximum immobilization yield of 24.7%. Optimum pH for the free and immobilized enzyme was pH 8 and 9, respectively. However, no change was observed in optimum temperature ($50^{\circ}C$). Thermal stability was observed in immobilized enzymes compared to free enzymes. The immobilized enzyme retained 86% activity after 10 cycle operations in a repeated batch process. The effect of flow rate on the stability of enzyme activity in continuous packed-bed reactor was investigated. Lowering flow rate increased the stability of the immobilized enzyme. After 96 hr of continuous operation in a packed-bed reactor, the immobilized enzyme retained 83 and 61% activity when casein and egg yolk were used as a raw materials, respectively.

Effect of Testosterone on Free Radical Generating Enzyme and Lipid Peroxidation (지질과산화 반응과 Free Radical 생성계 효소활성에 미치는 Testosterone의 영향)

  • Huh, Keun;Shin, Uk-Seob;Park, Jong-Min
    • YAKHAK HOEJI
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    • v.38 no.2
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    • pp.166-173
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    • 1994
  • Sex hormones not only regulate external sexual characteristics but several internal biochemical processes. It is well accepted that life-span of female is longer than that of male. Life-span is closely related with aging process in which free radicals are known to be involved. We investigated the effect of testosterone on free radical generating systems and lipid peroxidation based on the sexual difference. Lipid peroxide levels of male and female mouse were increased proportionately with age, especially in male mouse. Increase in enzyme activity of aldehyde oxidase with age was observed in male mouse, while no siginificant change in enzyme activity was found in female mouse. Enzyme activity of xanthine oxidase also showed similar results. It, however, was not significant statistically. Lipid peroxide level and xanthine oxidase type conversion ratio of male and female mouse liver homogenate incubated at $37^{\circ}C$, increased remarkably in proportion to incubation time, especially in male mouse. Lipid peroxide level and aldehyde oxidase activity were measured in normal male mouse, castrated mouse and testosterone treated-castrated mouse. Castrated mouse group showed decrease in lipid peroxide level and aldehyde oxidase activity compared with normal group. Treatment of castrated mouse with testosterone, however turned the level of lipid peroxide and aldehyde oxidase activity back to normal. From the above results, it might be concluded that testosteron could increase the activities of free radical generating enzymes which would result in the formation of lipid peroxide, consequently leading to aging.

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Kinetic Studies on Enzymatic Hydrolysis of Cellulose(I) -Effect of Structural Features of Cellulose on Enzyme Adsorption- (섬유소 가수분해반응에 관한 연구(I) -효소흡착에 대한 섬유소의 구조적 특성-)

  • Lee, Yong-Hun;Kim, Chul
    • KSBB Journal
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    • v.6 no.2
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    • pp.157-166
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    • 1991
  • The structural properties of cellulose are significantly changed with the progress of hydrolysis reaction. The effects of changes on such properties of cellulosic substrate as crystallinity, amicessibility of enzyme to the active site of cellulose surface, and particle size on the kinetics of enzymatic hydrolysis have been studied. Among those physical studies, the apparent surface active site of cellulose particle was found to have the most significant effect on the hydrolysis kinetics. Based on the experimental results, the adsorption affinity of enzyme and hydrolysis rate were mainly influenced by the surface roughness of cellulose particle. The extent of accesssible active site may be expressed as the change of particle diameter. The Langmuir isotherm was proposed in terms of enzyme activity to explain the actual action of enzyme protein.

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Activity Change of Ornithine Decarboxylase(ODC) after Hepatectomy

  • Chang Sung;Jou, Kab-Yeo;Ha, Duk-Mo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.24 no.3
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    • pp.415-419
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    • 1995
  • Ornithine decarboxylase(ODC) catalyzes the first and key step in the polyamine biosynthetic pathway. Ornithine decarboxylase is known to the enzyme that increase substantially its activity in regenerating liver. We found that activity and mRNA level for ODC increase significantly after partial hepatectomy in the rat. After laparotomy, there was significant decrease in activity ; however, mRNA content was unaltered in contrast to previous reports of no change in ornithine decarboxylase and thymidine kinase after sham hepatectomy. This may be mediated by the decrease in food intake after hepatectomy. Therefore it is necessary to examine the effect of food intake after hepatectomy on the ODC activity and mRNA level in the future.

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