• Title/Summary/Keyword: non-enzymatic activity

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Biotransformation of Pregnane Glycosides from Cynanchum wilfordii Roots by β-Glucosidase (당 분해효소를 이용한 백하수오 뿌리로부터 분리한 Pregnane Glycosides의 생전환)

  • Yoon, Mi-Young;Cuong, Mai Nguyen;Choi, Gyung-Ja;Choi, Yong-Ho;Jang, Kyoung-Soo;Cha, Byeong-Jin;Kim, Jin-Cheol
    • Research in Plant Disease
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    • v.18 no.3
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    • pp.186-193
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    • 2012
  • Biotransformation is an eco-friendly and efficient method for enhancing the bioavailability of biopesticide. To increase the antifungal activity of the crude extract of Cynanchum wilfordii roots against barely powdery mildew, we performed biotransformation of wilfoside C1G using ${\beta}$-glucosidase (cellobiase from Aspergillus niger). The mixture (G sample) of partially purified wilfoside C1G and cynauricuoside A (K1G) was treated with ${\beta}$-glucosidase to remove a glucopyranosyl moiety. The enzyme completely converted C1G to C1N and K1G to K1N. Optimal conditions for enzymatic biotransformation of G sample were determined to be 10% ethanol, 1,555 ${\mu}U$ ${\beta}$-glucosidase/ml, pH 5, and $45^{\circ}C$. In in vivo experiment, the G sample transformed by ${\beta}$-glucosidase showed stronger antifungal activity against barley powdery mildew than the non-treated G sample. These results suggest that ${\beta}$-glucosidase biotransformation can be applied to increase the antifungal activity of the crude extract of C. wilfordii roots against powdery mildews.

Electrocatalytic activity of the bimetallic Pt-Ru catalysts doped TiO2-hollow sphere nanocomposites (Pt-Ru@TiO2-H 나노구조체촉매의 합성 및 전기화학적 특성평가)

  • Lee, In-Ho;Kwen, Hai-Doo;Choi, Seong-Ho
    • Analytical Science and Technology
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    • v.26 no.1
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    • pp.42-50
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    • 2013
  • This paper describes the electrocatalytic activity for the oxidation of small biomolecules on the surface of Pt-Ru nanoparticles supported by $TiO_2$-hollow sphere prepared for use in sensor applications or fuel cells. The $TiO_2$-hollow sphere supports were first prepared by sol-gel reaction of titanium tetraisopropoxide with poly(styrene-co-vinylphenylboronic acid), PSB used as a template. Pt-Ru nanoparticles were then deposited by chemical reduction of the $Pt^{4+}$ and $Ru^{3+}$ ions onto $TiO_2$-hollow sphere ($Pt-Ru@TiO_2-H$). The prepared $Pt-Ru@TiO_2-H$ nanocomposites were characterized by transmission electron microscopy (TEM), X-ray diffraction (XRD), and elemental analysis. The electrocatalytic efficiency of Pt-Ru nanoparticles was evaluated via ethanol, methanol, dopamine, ascorbic acid, formalin, and glucose oxidation. The cyclic voltammograms (CV) obtained during the oxidation studies revealed that the $Pt-Ru@TiO_2-H$ nanocomposites showed high electrocatalytic activity for the oxidation of biomolecules. As a result, the prepared Pt-Ru catalysts doped onto $TiO_2$-H sphere nanocomposites supports can be used for non-enzymatic biosensor or fuel cell anode electrode.

Effects of Water Activity on the Non-Enzymatic Browning Reaction of Dry Milk (수분활성도(水分活性度)가 분유(粉乳)의 비효소적(非酵素的) 갈변(褐變)에 미치는 영향(影響))

  • Chang, Kyu-Seob;Chang, Yeong-Il;Chun, Ki-Chul;Lee, Seong-Gu
    • Applied Biological Chemistry
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    • v.28 no.4
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    • pp.261-270
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    • 1985
  • This study was carried out he effects of water activity on the nonenzymatic browning reactions of dry milk powders. Samples (Dry Whole Milk DWM, Nonfat Dry Milk NFDM) stored at $55^{\circ}C$ were analyzed for $O_2,\;CO_2$ and $H_2O$ by Gas chromatographic method. The brown color which developed from the Maillard reaction was also measured by a reflective spectrophotometer. And linear regression equations and the correlation coeffcients were determined for $O_2,\;CO_2$ and brown color vs. storage, and the results are as follows. The amount of brown and off-flavor development increase as the water activities increase about 0.44 Aw during storage. Both DWM and NFDM milk products show a plateau in water absorption between the 0.33 and 0.44 water activities. DWM produced more browning than NFDM which may be partially due to the carbonyl amine reaction. The $CaCl_2$ solution may exude a water activity lower than 0.44 and varies greatly with temperatures. Oxygen is depleted as brown color development.

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Studies on the Antibacterial Activity of Enzymatic Hydrolyzates of Lactoferrin Derived from Bovine Colostrum (유우 Lactoferrin 효소가수분해물 항균 활성에 관한 연구)

  • Han, Su Yeon;Kim, Jong Woo
    • Korean Journal of Agricultural Science
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    • v.25 no.1
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    • pp.52-67
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    • 1998
  • The investigative research on the mammalian milk purely consisted of the physiological quality of lactoferrin was conducted to reveal the antimicrobial ativity of specifically functional foods with antibiotic characteristics as a basic data in food manufacturing. Bovine lactoferrin were isolated from raw milk samples, and was digested with pepsin, trypsin and chymotrypsin. It was necessary then to separate and purify lactoferrin from bovine raw milk, and in order to analyze the antimicrobial activity of the enzyme-treated bovine lactoferrin in their required quantitative fraction. Afterwards Escherichia coli and Staphylococcus aureus was incubated in it. It was that investigated to enzyme-treated fractions molecular weight and the peptide fragment with antimicrobial effect. 1. The purity of enzyme-treated bovine lactoferrin(BLF) was tested by SDS-PAGE. As a results of 12% SDS-PAGE assay, pepsin-treated LF did not exhibited band until if reaches 14 KDa, while trypsin and chymotrypsin treated LF, known to contain the non-digestive lactoferrin exhibited band at a molecular weight of 33 KDa. 2. Bovine lactoferrin was sucessfully purified through the use of Sephadex G-50 Column. In order to assay LF through the Sephadex G-50 column chromatography, the digestive bovine lactoferrin (BLFs) was eluted with a linear gradient of 0.05% Tris-HCl. When the gel-filtration analysis, pepsin, trypsin and chymotrypsin treatments of BLF fragments was showed 2, 3, and 2 peak, respectively. The results of the HPLC analysis confirmed that had a non-digestive lactoferrin receptor, and trypsin and chymotrypsin treated BLFs has an antimicrobial effect. 3. To measure the strength of the antimicrobial effect of enzyme treated lactoferrin it was compared to the antimicrobial activity taking place at the incubated Escherichia coli and Staphylococcus aureus. This might explain the resistance of the microorganisms for peptide fragment. The pepsin-treated of bovine lactoferrin was markedly reduced by incubation of the cells. Trypsin-treated of BLF was similar to chymotrypsin-treated of BLF. However, trypsin and chymotrypsin treatments of BLFs were showed the antimicrobial effect until eight hours incubation for native bovine lactoferrin. Therefore the enzyme-treated lactoferrin have an antimicrobial effect even non-digestive lactoferrin. 4. The digestive bovine lactoferrin fragments assay was carried out by the use of Sephadex G-50 column chromatography and SDS-PAGE. The pepsin and chymotrypsin-treated fragments has a low molecular weight and trypsin-treated lactoferrin was only showed a band. It was described that characteristics of digestive protein. It appeared that there may be a relation between virulence and resistance to enzyme-treated BLF.

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Improvement on Yield of Extracts from Byproducts of Alaska Pollock Theragra chalcogramma and Sea Tangle Laminaria japonica using Commercial Enzymes and Its Food Component Characterization (상업적 효소를 이용한 명태(Theragra chalcogramma) 두부 및 정형 다시마(Laminaria japonica) 부산물 유래 고압 추출물의 수율개선 및 이의 식품성분 특성)

  • Noh, Yuni;Park, Kwon Hyun;Lee, Ji Sun;Kim, Hyeon Jeong;Kim, Min Ji;Kim, Ki Hyun;Kim, Jeong Gyun;Heu, Min Soo;Kim, Jin-Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.46 no.1
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    • pp.37-45
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    • 2013
  • This study was conducted to improve the yield of extracts from Alaska pollock Theragra chalcogramma head and sea tangle Laminaria japonica byproducts using various commercial enzymes, such as Alcalase, Flavourzyme, Neutrase (NH), and Protamex. Among the enzymatic hydrolysates, the yield was highest in hydrolysate incubated with NH for 4 h. NH-treated hydrolysates (NHH) also improved functional properties, such as angiotensin-I converting enzyme (ACE) inhibitory activity and 2,2-diphenyl-1-picryldrazyl (DPPH) radical scavenging activity, as compared to extracts from Alaska pollock head and sea tangle byproducts. Total free amino acid and taste values of NHH were 379.7 mg/100 mL and 24.03, respectively, after digestion for 4 h. These values are 2.2-fold and 1.9-fold higher compared with those of water soluble fractions extracted from Alaska pollock head and non-forming sea tangle, respectively. According to the taste value results, the major taste-active compounds among free amino acids of NHH were glutamic acid and aspartic acid. These results suggest that NHH can be used as an ingredient for natural seasoning preparation.

Effects of Houttuynia Cordata thunb on the liver damage of TCDD-treated rats (TCDD를 투여한 rat의 간손상에 대한 어성초의 효과)

  • 하배진;하종명;이상현;이재화;정혜진;이상헌;김희진;이진영
    • Journal of Life Science
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    • v.13 no.4
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    • pp.457-462
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    • 2003
  • Houttuynia Cordata thunb has been used as folk medicine for analgesics, beriberi, edema, hepatitis and icterus etc. We investigated, the effects of Houttuynia Cordata thunb administration on protective in liver of 2,3,7,8-tetrachlorodibenzo-p-dioxin(TCDD) treated rats. Seven days after the injection of TCDD(1${\mu}g$/kg), Houttuynia Cordata thunb (200mg/kg) was administered into rats intraperitoneally for four weeks. We examined the antioxidative enzymatic activity by measuring the level of GOT, GPT in serum and MDA, GSH, GSSG, GPx, SOD and Catalase in liver tissue of rats. GOT activity of Houttuynia Cordata thunb and TCDD administered group(HTT) showed 49.00% of inhibitive effect compared to TCDD-treated abnormal group(TTA). GPT level of HTT group was decreased to the level of Non TCDD-treated group(NTT). MDA content in the TTA group was 1.27 times increased compared to NTT group. HTT group was inhibited by 69.53% compared to TTA group. GSH contents in HTT group was 1.91 times increased compared to TTA group. GSSG contents in HTT group was 46.72% decreased compared to TTA group. SOD and Catalase in TTA group were lower than in NTT group, but SOD and Catalase in HTT group were increased by 82% and 55.45% respectively compared to TTA group.

Effect of the Inhibition of PLA2 and PAF on the Neutrophilic Respiratory Burst and Apoptosis (호중구의 Respiratory Burst에 미치는 PLA2 및 PAF와 영향 : In vitro에서의 호중구의 산소기 생성 및 Apoptosis에 관한 연구)

  • Lee, Young-Man;Kim, Sang-Gyung;Park, Yoon-Yub
    • Tuberculosis and Respiratory Diseases
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    • v.48 no.6
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    • pp.887-897
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    • 2000
  • Background : Since the exact pathogenesis of sepsis-induced ARDS has not been elucidated, the mechanisms of enhanced neutrophilic respiratory burst were probed in endotoxin primed neutrophils associated with the roles of phospholipase A2(PLA2), platelet activating factor(PAF) and apoptosis. Methods : In isolated fresh human neutrophils, effects of the inhibition of PLA2 and PAF on the apoptosis were examined by the method of Annexin-FITC/dual PIflow cytometry. The roles of PLA2 and PAF on the neutrophilic respiratory burst were also examined by measuring oxidant generation in cytochrome-c reduction assay. Activities of the PLA2 and lysoPAF acetyltransferase (lysoPAF AT) of the neutrophils were determined to understand the effect of endotoxin on these enzymatic activities which may be related to the neutrophilic respiratory burst and apoptosis. In addition, the role roles of PLA2 and PAF in neutrophilic adhesion to bovine endothelial cells were examined in vitro by neutrophil adhesion assay. To investigate the effect of oxidants on pulmonary surfactant, cytochemical ultrastructural microscopy was performed. To inhibit PLA2 and PAF, non-specific PLA2 inhibitor mepacrine (100 nM) and WEB 2086 (100 nM) or ketotifen fumarate (10 ${\mu}g$/ml) were used respectively in all in vitro experimental sets. WEB 2086 is PAF receptor antagonist, and ketotifen fumarate is a lyso PAF AT inhibitor. Results: The mapacrine treatment, provided and the endotoxin (ETX) treatment, resulted in increased apoptosis of neutrophils (p<0.001) while treatments of WEB 2086 and ketotifen did not. The inhibition of PLA2 and PAF decreased (p<0.001) production of oxidants from PMA-stimulated neutrophils. While endotoxin increased the PLA2 activity of neutrophils (p<0.01), mepacrine supressed (p<0.001) the activity, provided after treatment of ETX. The lyso PAF actyltransferase activity (lyso PAF AT) increased (p<0.01) after treatment of ETX. In contrast, mepacrine, WEB 2086 and ketotifen showed a tendency of decreasing the activity after treatment of ETX. The treatment of ETX incresed (p<0.001) neutrophil adhesion to endothelial cells, which was reversed by inhibition of PLA2 and PAF (p<0.01). The binding of oxidants to pu1monary surfactant was identified histologically. Conclusions : The enhanced neutrophilic respiratory burst by ETX plays a pivotal role in the pathogenesis of ARDS in terms of oxidayive oxidative stress. Increased production of oxidants from neutrophils is mediated by the activations of PLA2 and lyso PAF AT.

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Polymorphic Variation in Glutathione-S-transferase Genes and Risk of Chronic Myeloid Leukaemia in the Kashmiri Population

  • Bhat, Gulzar;Bhat, Ashaqullah;Wani, Aadil;Sadiq, Nida;Jeelani, Samoon;Kaur, Rajinder;Masood, Akbar;Ganai, Bashir
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.1
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    • pp.69-73
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    • 2012
  • Cancer is a complex disease and the genetic susceptibility to it could be an outcome of the inherited difference in the capacity of xenobiotic metabolizing enzymes. Glutathione-S-transferases (GSTs) are phase II metabolizing enzymes whose various genotypes have been associated with increased risk of different types of cancer. Null mutations caused by the deletion of the entire gene result in the absence of the enzymatic activity and increase in the risk of developing cancer including chronic myeloid leukaemia (CML). In the present case-control study we evaluated the effect of null mutations in GSTM1 and GSTT1 genes on the risk of developing CML. The study included 75 CML patients (43 males and 32 females; age (mean ${\pm}$ S.D) $42.3{\pm}13.4$ years) and unrelated non-malignant controls (76 male and 48 females; age (mean ${\pm}$ S.D) $41.5{\pm}12.9$). The distribution of GSTM1 and GSTT1 genotypes in CML patients and controls was assessed by multiplex-PCR method. Logistic regression was used to assess the relationship between GSTM1 and GSTT1 genotypes and risk of CML. Chi-square test was used to evaluate the trend in modulating the risk to CML by one or more potential high risk genotype. Although GSTM1 null genotype frequency was higher in CML patients (41%) than in the controls (35%), it did not reached a statistical significance (OD = 1.32, 95% CI: 0.73-2.40; P value = 0.4295). The frequency of GSTT1 null genotypes was higher in the CML patients (36%) than in the controls (21%) and the difference was found to be statistically significant (OD = 2.12, 95% CI: 1.12-4.02; P value = 0.0308). This suggests that the presence of GSTT1genotype may have protective role against the CML. We found a statistically significant (OD = 3.09, 95% CI: 1.122-8.528; P value = 0.0472) interaction between the GSTM1 and GSTT1 null genotypes and thus individuals carrying null genotypes of both GSTM1 and GSTT1 genes are at elevated risk of CML.

Chaperone Therapy in Gaucher Disease (고셔병에서 샤프론 치료)

  • Lee, Beom Hee;Heo, Sun H.;Cheon, Chong Kun;Kim, Yoo-Mi;Kim, Ja Hye;Choi, In Hee;Kim, Jae-Min;Kim, Gu-Hwan;Yoo, Han-Wook
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.13 no.1
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    • pp.37-42
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    • 2013
  • Gaucher disease is a lysosomal storage disease caused by deficiency of glucocerebrosidase (GBA). This condition is characterized by accumulation of glucocerebrosidase in liver, spleen, lung, skeletal system, and central nervous system. Gaucher disease is the prototype of disease in which efficacy of enzyme replacement therapy has been established. However, because recombinant enzyme is not able to enter the central nervous system, its efficacy is limited to the non-neurological manifestations of Gaucher disease. Importantly, approximately a half of Korean patients with Gaucher disease suffer from neurological manifestations. In addition, Korean Gaucher disease patients exhibit distinct mutation spectrum from those in other populations. Common mutations in Korean patients with Gaucher disease are also associated with neurological phenotype. Therefore, therapeutic strategies tailored to Korean patients were necessary. Interestingly, a chemical chaperone, ambroxol, has been known to increase residual enzymatic activities of the select mutant GBAs encoded by mutations prevalent in Korean patients. One promising aspect of this drug is that it can cross blood-brain barrier, and enhance the enzyme activity in the brain. In vitro study suggested this chemical chaperone as one of new therapeutic agents in Gaucher disease, and a well-designed human trial is required to confirm its efficacy.

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The Non-Enzymatic Browning and Shelf-Life of Dried Shrimp during Storage under Fluctuating Temperature Conditions (건조 새우의 변온저장중 갈변 및 Shelf-life)

  • KIM Yong-Ju;KIM Mu-Nam;KANG Moon-Sun;CHO Young-Je;KIM Yuck-Yong;CHUN Soon Sil
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.27 no.1
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    • pp.7-12
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    • 1994
  • The kinetics of browning reaction of dried shrimp powder samples were investigated during storage. The temperature conditions of the ding process were $25^{\circ}C\;and\;45^{\circ}C$, and the samples were stored at water activity ($a_w$) of 0.33, 0.44, 0.52, 0.65 and temperatures of $35^{\circ}C,\;45^{\circ}C,\;55^{\circ}C$ and temperature fluctuations between $33^{\circ}C\;and\;55^{\circ}C$. When the shrimp was dried at $25^{\circ}C$ the activation energies obtained from the Arrhenius plot ranged from 13.57 to 14.33 kcal/mol. From these energies of activation, the $Q_{10}$ values at $25^{\circ}C$ showed 1.93 to 2.00. In the case of drying at $45^{\circ}C$ the activation energies were $13.12{\sim}13.61\;kcal/mol$ and $Q_{10}$ values were $1.89{\sim}1.93$, respectively. In addition, a storage study under square-wave fluctuating temperature conditions was carried out by varying the shrimp sample temperature between $35^{\circ}C\;and\;55^{\circ}C$ within 7 days regular fluctuation cycle. The data obtained from the fluctuating temperature storage study will be used in the prediction of shelf-lives. The shelf-lives assessed at $25^{\circ}C$ from the accelerated shelf-life tests ranged from 4 days at aw 0.65 to 139 days at aw 0.33.

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