• Title/Summary/Keyword: enzyme property

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Purification and Characterization of Urushiol Induced Laccase Isoenzyme from Fomitella fraxinea (Urushiol에 의해 유도된 장수버섯 laccase isoenzyme의 정제 및 특성)

  • Choi, Han-Seok;Park, Hyo-Suk;Yeo, Soo-Hwan;Jeong, Seok-Tae;Choi, Ji-Ho;Kim, Myung-Kon
    • The Korean Journal of Mycology
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    • v.38 no.2
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    • pp.152-159
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    • 2010
  • The influence of urushiol, as an allergen on laccase property of Fomitella fraxinea was investigated. The enzyme production was reached to the highest level after 10 days, cultivation and the activity and mycelial biomass were increased by 2.5 and 1.5 folds, respectively, by adding urushiol in the culture medium. In liquid cultures using a Cu Mn-free medium, laccase lactivity was decreased by 3.8-9.2 folds, with similar dry cell weight. Two isoenzymes, were purified using anion exchange, hydrophobic interaction and size-exclusion chromatographies. Both isoenzymes are monomeric proteins, with $M_W$ around 67 kDa(Lac1) and 66 kDa(Lac2), and isoelectric points of 3.67 and 3.81. The optimal conditions for purified isoenzymes were found to be pH 4.5-5.0 and $30-35^{\circ}C$. Activity decreased by the addition of $Fe^{2+}$, $Mg^{2+}$, $Na^+$, and strongly inhibited by EDTA and sodium azide.

Recent Progress in Colorimetric Assays Using the Absorption of Plasmonic Gold Nanoparticles (플라즈모닉 금 나노입자의 흡광 특성을 활용한 생화학적 비색 분석법 연구 동향)

  • Bong-Geun Kim;Sang Bin Yoon;Sukyeong Hwang;Hyon Bin Na
    • Applied Chemistry for Engineering
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    • v.35 no.2
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    • pp.67-78
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    • 2024
  • Light absorption has potential as a signal in biochemical analyses due to its simplicity in measurement and interpretational clarity. Among substances that generate absorption signals, gold nanoparticles possess advantages such as chemical stability, biological compatibility, and unique optical properties from the localized surface plasmon resonance (LSPR) in the visible light range. They also exhibit versatility compared to other colorimetric substances effective only for specific target molecules, as they easily conjugate with various detection active substances like antibodies and aptamers. Particularly due to advantages such as low cost, ease of particle synthesis, and high environmental stability compared to enzyme-based colorimetric methods, gold nanoparticles are extensively researched as signal substances in colorimetric assays. This review summarizes various strategies utilizing gold nanoparticles as absorption signal substances, focusing on recent research. Based on the characteristics of gold nanoparticles, where the optical property is influenced by particle morphology, literature is classified and reviewed based on strategies controlling the shape of gold nanoparticles during signal generation. Through this, it is observed that gold nanoparticles, which have been used as absorption signal substances, continue to be actively researched, affirming their potential for broad and continuous improvement in the future.

Enzymatic Synthesis of Flame Retardant Phenolic Polymers Catalyzed by Horseradish Peroxidase (Horseradish Peroxidase 효소촉매에 의한 난연성 페놀고분자의 합성)

  • Park, Han Sol;Park, Jung Hee;Lee, Hak Sung;Ryu, Keungarp
    • Korean Chemical Engineering Research
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    • v.51 no.1
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    • pp.111-115
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    • 2013
  • The optimum synthetic conditions of poly(p-phenylphenol) by horseradish peroxidase in dioxane:water (80:20 v/v) mixtures were studied. The stability against thermal degradation and structural properties of the synthesized phenolic resins were investigated by thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC), respectively. The synthetic yield of poly(p-phenylphenol) increased upon the increase of the amount of enzyme up to 0.25 mg HRP/mL, then leveled off for further increase of the enzyme usage. When sodium acetate (100 mM, pH 4~6) and sodium phosphate (100 mM, pH 7~9) were used as the buffering salts for the aqueous component (20% v/v), the synthetic yield of the resin increased at higher pH of the aqueous buffer. But when the pHs of the aqueous buffer were 6 and 9, the synthetic yield strongly depended on the types of the buffering salts; if sodium phosphate was used instead of sodium acetate at pH 6, the yield decreased by about 15% and if sodium bicarbonate was used instead of sodium phosphate, the yield decreased by almost 20%. When the pH range of the aqueous buffer was from 4 to 7, the addition of a radical mediator, 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonate) (ABTS), up to 2 mM improved the synthetic yield of the resin by about 10%. TGA experiments revealed that the thermal stability of the resin synthesized in dioxane:water (100 mM sodium phosphate, pH 9) (80:20 v/v) was high having the char yield of 47% upon the heating at $800^{\circ}C$. DCS results showed that the structures of the polymers synthesized in acidic aqueous buffers were different from those of the polymers synthesized in the basic aqueous buffers. However, all the synthesized resins were found to have the property of the thermosetting resins.

Purification and Biological Characterization of Wild-type and Mutants of a Levan Fructotransferase from Microbacterium sp. AL-210 (Microbacterium sp. A-210이 생성하는 Levan fructotransferase의 정제 및 생물학적 특성에 관한 연구)

  • Hwang, Eun-Young;Jeong, Mi-Suk;Cha, Jae-Ho;Jang, Se-Bok
    • Journal of Life Science
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    • v.19 no.9
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    • pp.1218-1225
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    • 2009
  • Difractose anhydrides (DFAs) is studied as a sweetener for diabetics because of its structural property. DFAs have four types: DFA I, III, IV (degradation of levan) and V (degradation of inulin). Especially, DFA IV has been shown to enhance the absorption of calcium in experiments using rats. Levan fructotransferase is an enzyme for producing di-d-fructose-2,6':6,2-dianhydride (DFA IV). To identify structural characterization, we purified wild-type and mutants (D63A, D195N and N85S) of levan fructotransferase (LFTase) from Microbacterium sp. AL-210. These proteins were purified to apparent homogeneity by Ni-NTA affinity column, Q-sepharose ion exchange and gel filtration chromatography and detected by SDS-PAGE. They were also analyzed by circular dichroism (CD) measurements, JNET secondary structure prediction, activity measurements at various temperatures, and pH analysis. The optimum pH for the enzyme-catalyzed reaction was pH 7.5 and optimum temperature was observed at $55^{\circ}C$. Along with wild-type LFTase, mutants were analyzed by CD measurement, fluorescence analysis and differential scanning calorimetry (DSC). N85S showed less $\alpha$-helix and more $\beta$ strand than others. Also, N85S showed almost the same curve as wild-type in their steady-state fluorescence spectra, whereas mutant D63A and D195N showed higher intensity than wild-type. The amino acid sequence of wild-type LFTase was compared to the sequences of exo-inulinase from Aspergillus awamori, a plant fructan 1-exohydrolase from Cichorium intybus, and Thermotogo maritime (Tm) invertase and showed a high identity with Exo-inulinase from Aspergillus awamori.

Quality characteristics of Takju according to different rice varieties and mixing ratio of Nuruk (쌀 품종과 누룩 배합비율에 따른 탁주의 품질 특성)

  • Im, So-Yeon;Baek, Chang-Ho;Baek, Seong-Yeol;Park, Hye-Young;Choi, Han-Seok;Choi, Ji-Ho;Jeong, Seok-Tae;Shin, Woo-Chang;Park, Heui-Dong;Yeo, Soo-Hwan
    • Food Science and Preservation
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    • v.21 no.6
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    • pp.892-902
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    • 2014
  • The quality characteristics of Takju were investigated according to the different rice varieties and mixing ratio of Nuruk for the advanced quality of Takju. The yeast was selected by alcohol-producing ability. Then a liquid starter was prepared using brewing fungi (Aspergillus luchuensis 34-1 and Lichtheimia ramosa CN042), and rice Nuruk was manufactured with two rice types (Chucheong and Hanareum). The quality characteristics of Takju were investigated based on the rice type and the mixing ratio of A. luchuensis 34-1 and Lich. ramosa CN042 (1:0, 0:1, 1:1, 1:3). S. cerevisiae Y268 showed an alcohol yield of $9.3{\pm}0.33%$ at a 0.3% concentration in the YPD broth medium, and the rice Nuruk with A. luchuensis 34-1, regardless of rice type, was confirmed to have a higher enzyme activity and physiochemical property than Lich. ramosa CN042. According to the quality analysis of Takju, the physiochemical property was increased for the fermentation period, and the acidity differed by type of fungi and rice. The quality of Takju was changed with the composition differences of organic acid and free amino acid by rice type and mixing ratio. As a result of the sensory evaluation of Takju, the preference for it was increased with the Hanareum and A. luchuensis 34-1, respectively. Thus, this study shows the possibilities for activating the industry of traditional liquor by improving the Nuruk and Takju manufacturing technique.

Inhibitory Effects on the Enzymes Involved in the Inflammation by the Ethanol Extracts of Plant Foodstuffs (식물성 일반식품 자원의 에탄올 추출물이 염증 효소계에 미치는 영향)

  • Kwon, Eun-Sook;Kim, Il-Rang;Kwon, Hoon-Jeong
    • Korean Journal of Food Science and Technology
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    • v.39 no.3
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    • pp.348-352
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    • 2007
  • Inflammation is a complex process resulting from a variety of mechanisms. Combined inhibition of the activities of enzymes involved in the process may therefore be considered more important in anti-inflammatory property of plant extracts than any single contribution. In this study, the inhibitory effects of the ethanol extracts of thirty plant foods on the activities of secretory phospholipase $A_{2}$ ($sPLA_{2}$), cyclooxygenase-1 (COX-1), cyclooxygenase-2 (COX-2), and 12-lipoxygenase (12-LOX) were examined. Several legumes, mungbean sprout and some leaf vegetables inhibited the activity of $sPLA_2$, upstream enzyme of inflammation pathway. Only soybean sprout and mungbean sprout significantly inhibited 12-LOX activity. Although most of extracts inhibited the activities of both COX-1 and COX-2, water dropwort and amaranth showed selectivity for the inhibition of COX-2 over COX-1. Especially, mungbean showed anti-inflammatory property at both upstream and downstream of inflammation pathway with relatively low $IC_{50}$ values for $sPLA_{2}$ and COX-2 enzymes. Mungbean sprout exhibited inhibitory effects on all enzymes related to early and late inflammation and soybean sprout suppressed 12-LOX and COX-2 simultaneously, although the activities of these plants were showed at relatively high concentration. Therefore, mungbean, mungbean sprout, and soybean sprout appear to exhibit anti-inflammatory effects by combined inhibition of inflammatory enzymes.

Effects on Storage Property of Fresh Jujube Treated with Grape Fruit Stem Extracts (GFSE) and Fermented Pollen (FP) (포도줄기추출물 및 발효화분 처리가 생대추의 저장기간에 미치는 영향)

  • Woo, Koan-Sik;Hwang, In-Guk;Jang, Keum-Il;Kang, Tae-Su;Lee, Hee-Bong;Jeong, Heon-Sang
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.8
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    • pp.1090-1094
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    • 2008
  • Treatment effects of grape fruit stem extracts (GFSE) containing trans-resveratrol or fermented pollen (FP), and ethylene gas scavenging material (EGS) on the storage property of fresh jujube (Z izyphus jujuba forma hoonensis C.S. Yook) were investigated. Fresh jujubes were packed in different storage containers (PE film and Lock & Lock vessel), and treated with GFSE containing trans-resveratrol of 30 ppm or 1% FP, and EGS. The storage vessels were stored in refrigerator ($0{\pm}1^{\circ}C$) during 12 weeks, and then quality characteristics during storage period were analysed. Hardness slightly increased until 4 weeks and decreased afterward. Soluble solid ($^{\circ}Bx$) and total titratable acidity of fresh jujube slightly increased in all treatments during storage period. Vitamin C content of fresh jujube slightly decreased in all treatments during storage period. Number of microorganisms decreased until 4 weeks and increased afterward. The decay enzyme activity increased in all treatments during storage period. Storage stability was higher for PE film than Lock & Lock containers. Storage period of fresh jujube in this experiment ranges in $8{\sim}9$ weeks for maturity fruits treated with 30 ppm of GFSE and ethylene gas scavenging material.

Studies on Rheological Characterization of Barley ${\beta}-Glucan$ [mixed-linked $(1-3),(1-4)-{\beta}-D-Glucan$] (보리 ${\beta}-Glucan$ [mixed-linked $(1-3),(1-4)-{\beta}-D-Glucan$의 리올로지 특성)

  • Kim, Mi-Ok;Cha, Hee-Sook;Koo, Sung-Ja
    • Korean Journal of Food Science and Technology
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    • v.25 no.1
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    • pp.15-21
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    • 1993
  • Crude ${\beta}-glucan$ extracted from Barley was purified by stepwise enzyme treatment (Thermostable ${\alpha}-amylase$, amyloglucosidase, protease). The Intrinsic Viscosity $[{\eta}]$ of the purified ${\beta}-glucan$ was determined by Cannon Fenske Capillary Viscometer (size 50, Cannon Instruments, State, College pa.) at different pH (2, 4, 7, 9, 11) and various salt concentration (0.01 M, 0.03 M, 0.05 M, 0.07 M, 0.1 M and 0.2 M). The $[{\eta}]$ of purified ${\beta}-glucan$ was ranged from $0.997{\sim}2.290\;dl/g$. The $[{\eta}]$ of purified ${\beta}-glucan$ at both alkali, acid condition were lower than those at pH 7. However, the alkali condition of puified ${\beta}-glucan$ solution showed less $[{\eta}]$ than the acid condition of this solution. From 0 M to 0.2 M salt concentration, the $[{\eta}]$ of purified ${\beta}-glucan$ solution was decreased to 0.03 M then increased to 0.05 M NaCl and remained constant to 0.2 M NaCl. The chain stiffness parameter of purified ${\beta}-glucan$ was not affected by temperature from $15^{\circ}C$ to $65^{\circ}C$. The shear rates of various ${\beta}-glucan$ conditions were determined by Bohlin Rheometer (Lund, Sweden). The ${\beta}-glucan$ concentration of 1.0 g/dl and 2.0 g/dl behaved as Newtonian fluid. However, above the concentration of 3.0 g/dl ${\beta}-glucan$ solution, it showed thixotropic and psedoplastic characteristics. Barley ${\beta}-glucan$ appears a damping at 0.5 frequency for the 4.0 g/dl solution. Below 0.5 frequency, it appears a viscous behavior property and above 0.5 frequency, it appears a elastic behavior property.

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An Investigation of Glyceollin I's Inhibitory Effect on The Mammalian Adenylyl (글리세올린 I의 아데니닐 고리화 효소 활성 억제 효능과 결합 부위 비교 분석)

  • Kim, Dong-Chan;Kim, Nam Doo;Kim, Sung In;Jang, Chul-Soo;Kweon, Chang Oh;Kim, Byung Weon;Ryu, Jae-Ki;Kim, Hyun-Kyung;Lee, Suk Jun;Lee, Seungho;Kim, Dongjin
    • Journal of Life Science
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    • v.23 no.5
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    • pp.609-615
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    • 2013
  • Glyceollin I has gained attention as a useful therapy for various dermatological diseases. However, the binding property of glyceollin I to the mammalian adenylyl cyclase (hereafter mAC), a critical target enzyme for the down-regulation of skin melanogenesis, has not been fully explored. To clarify the action mechanism between glyceollin I and mAC, we first investigated the molecular docking property of glyceollin I to mAC and compared with that of SQ22,536, a well-known mAC inhibitor, to mAC. Glyceollin I showed superiority by forming three hydrogen bonds with Asp 1018, Trp 1020, and Asn 1025, which exist in the catalytic site of mAC. However, SQ22,536 formed only two hydrogen bonds with Asp 1018 and Asn 1025. Secondly, we confirmed that glyceollin I effectively inhibits the formation of forskolin-induced cAMP and the phosphorylation of PKA from a cell-based assay. Long term treatment with glyceollin I had little effect on the cell viability. The findings of the present study also suggest that glyceollin I may be extended to be used as an effective inhibitor of hyperpigmentation.

The Efficacy Evaluation of Tourmaline-Ionized Water in Animal Study (투어마린이온활성수의 효능 평가)

  • Yoon, Yang-Suk;Kim, Dong-Heui;Qi, Xu-Feng;Song, Soon-Bong;Jung, Jong-Ho;Joo, Kyung-Bok;Teng, Yung-Chien;Lee, Kyu-Jae
    • Applied Microscopy
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    • v.39 no.4
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    • pp.311-317
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    • 2009
  • This study was performed using animals to confirm the effect of tourmaline-ionized water (TIW) the properties of which were changed by tourmaline energy and electric discharge. In the ICR mice fed high-fat diet, body weight increasing rate of the TIW-treated group (Exp) was generally decreased and moreover exhibited significance at 11th week (P<0.05) compared with the control (Con) group fed distilled water, although water intake of the Exp group was lower than that of the Con group. In the ICR mice with $CCl_4$-induced hepatotoxicity, AST and ALT activities of the Exp group were not significant but showed some decreasing trend, and histological damage of liver was less compared with thatof the Con group. On the study of ethanol-induced hangovers in Sprague-Dawley rat, blood alcohol concentration was significantly decreased (P<0.01), activity of GST, antioxidant enzyme related to the alcohol metabolism, was increased in liver tissue (P<0.05), and AST and ALT show a tendency to be decreasedin the Exp group. These results suggest that drinking TIWhas not only some obesity preventing effect but also an alcohol detoxification effect and liver protecting effect in vivo. It is supposed due to a structural change of water cluster and a property which maintains the changed structure through tourmaline energy and electric discharge. Therefore, TIW has a potentiality to be developed as functional water with several beneficial effects as well as for daily drinking, but further study on the mechanism related with efficacy will be necessary.