• Title/Summary/Keyword: Enzyme complex

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New Butyrylcholinesterase Inhibitory Triterpenes from Salvia santolinifolia

  • Mehmood Sajid;Riaz Naheed;Nawaz Sarfraz Ahmed;Afza Nighat;Malik Abdul;Choudhary Muhammad Iqbal
    • Archives of Pharmacal Research
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    • v.29 no.3
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    • pp.195-198
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    • 2006
  • Slavins A (1) and B (2), the new amyrin type triterpenes, have been isolated from the chloroform soluble fraction of Salvia santolinifolia and assigned structures on the basis of spectral studies including 2D NMR. Both the compounds displayed inhibitory potential against the enzyme butyrylcholinesterase.

Recovery of Cholesterol from the $\beta$-Cyclodexgtrin-Cholestrerol Complex Using Immobilized Cyclomaltodextrinas of Alkalophilic Bacillus sp. KJ 133

  • Kwon, Ho-Jeong;Jung, Hye-Jin;Kwak, Hae-Soo
    • Journal of Microbiology and Biotechnology
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    • v.11 no.4
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    • pp.712-715
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    • 2001
  • A new combined method including the enzymatic hydrolysis of $\beta$-cyclodextrin ($\beta$-CD) and solvent extraction fo cholesterol from the hydrolyzed mixture was developed to recover cholesterol from a $\beta$-CD-cholesterol complex prepared from dairy products, such as cream, milk, and cheese. Cyclomaltodextrinase (cyclomatodextrin dextrin hydrolase, EC 3.2.1.54, DCase_ prepared form alkalophilic Bacillus sp. KJ 133 hydrolyzed the $\beta$-DC of the $\beta$-CD-cholesterol complex, and then, free cholesterol was efficiently extracted from the hydrolyzed mixture by a nonpolar solvent such as ethyl acetate. To increase the stability of free CDase, immobilized CDase was developed using sodium alginate as a carrier. The immobilized CDase showed a high recovery yield of cholesterol in a time-dependent manner compared to the free CDase. A gas chromatography analysis showed that more than 70% of cholesterol was recovered from the $\beta$-DC-cholesterol complex of cream by the immobilized CDase, whereas only 3% and 29% of cholesterol were recovered when the solvent extraction and free CDase treatment were used, respectively. The cholesterol recovered can be used as a raw material for steroid synthesis. Furthermore, this method can be an efficient way to recover cholesterol or other organic compounds that are bound in a $\beta$ -DC-cholesterol or -organic compound complex.

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Structural Studies of Copper(II)-Hippuryl-L-histidyl-L-leucine(HHL) Complex by NMR Methods

  • Lee Seong-Ran;Jun Ji-Hyun;Won Ho-Shik
    • Journal of the Korean Magnetic Resonance Society
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    • v.10 no.1
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    • pp.115-125
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    • 2006
  • Hippuryl-L-histidyl-L-leucine(HHL) is widely used as a substrate of angiotensin converting enzyme(ACE) cleaving the neurotransmitter angiotensin(I) to the octapeptide angiotensin(II). The structure of the substrate molecules should provide information regarding the geometric requirements of the ACE active site. For the purpose of determination of in vivo reaction, metallo(Cu, Zn)-HHL complexes were synthesized and the degree of complex formation were identified by MALDITOF, ESI mass spectrometric analysis. Tn addition, the pH-dependent species distribution curves were obtained by potentiometric titration. Nitrogen atoms of imidazole ring and oxygen atom of caboxylate groups in the peptide chain were observed to be participated in the metal complex formation. After purification of complexes further structural characterization were made by utilizing UV-Vis, electrochemical methods and NMR. Complete NMR signal assignments were carried out by using 2D-spectrum techniques COSY, TOCSY, NOESY, HETCOR. A complex that two imidazole and carboxylate groups are asymmetrically participating to coordination mode was predicted to the solution-state structure of $Cu(II)-HHL_2$ based on $^{13}C-NMR$ signal assignment and NOE information.

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Effect of Dietary α-1,6-Galactosidase and β-1,4-Mannanase on Growth Performance and Nutrient Digestibility in Nursery and Growing Pigs (자돈 및 육성돈에 있어 α-1,6-galactosidase와 β-1,4-mannanase의 사료내 첨가가 성장 및 영양소 소화율에 미치는 영향)

  • Kwon, O. S.;Kim, I. H.;Lee, S. H.;Hong, J. W.;Kim, J. H.;Moon, T. H.;Lee, J. H.
    • Journal of Animal Science and Technology
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    • v.45 no.2
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    • pp.211-218
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    • 2003
  • For the Exp. 1, a total of sixty pigs (10.57$\pm$0.30kg average initial body weight) were used in a 15-d growth assay to determine the effect of dietary $\alpha$-1,6-galactosidase and $\beta$-1,4-mannanase on growth performance and nutrient digestibility. Dietary treatments included 1) CON (corn-dried whey-SBM based diet), 2) EC0.1 (CON diet+0.1% enzyme complex of $\alpha$-1,6-galactosidase and $\beta$-1,4-mannanase). Through the entire experimental period, gain/feed of pigs fed EC0.1 diet was higher (0.43 vs 0.52) than that of pigs fed CON diet (P<0.05). Pigs fed EC0.1 diet showed significant (P<0.05) improvement in dry matter (74.82% vs 82.41%) and nitrogen (70.59% vs 77.88%) digestibilities compared to pigs fed CON diet. For the Exp. 2, a total of thirty six pigs (22.30$\pm$0.45kg average initial body weight) were used in a 30-d growth assay to determine the effects of dietary $\alpha$-1,6-galactosidase and $\beta$-1,4-mannanase in low energy diet on growth performance and nutrient digestibility. Dietary treatments included 1) AME (adequate ME diet), 2) AME+EC0.1 (AME diet+0.1% enzyme complex) and LME+EC0.1 (low ME diet + 0.1% enzyme complex). Through the entire experimental period, average daily feed intake of pigs fed enzyme complex supplemented diets was higher than that of pigs fed CON diet (P<0.05). Also, pigs fed AME+EC0.1 diet showed significant (P<0.05) increase in ADFI (1,401g vs 1,733g) compared to pigs fed CON diet. Pigs fed enzyme complex supplemented diet showed significant (P<0.05) improvement in dry matter and nitrogen digestibilities compared to pigs fed CON diet. In conclusion, the results obtained from these feeding trials suggest that the supplementation of $\alpha$-1,6-galactosidase and $\beta$-1,4-mannanase was an effective means for improving growth performance and dry matter and nitrogen digestibilities in nursery and growing pigs.

A single-center, randomized, double-blind, placebo-controlled study on the efficacy and safety of "enzyme-treated red ginseng powder complex (BG11001)" for antiwrinkle and proelasticity in individuals with healthy skin

  • Park, Sang-Yong;Shin, Yu-Kyong;Kim, Hee-Taek;Kim, Yong Min;Lee, Don-Gil;Hwang, Eunson;Cho, Byung-Goo;Yin, Chang Shik;Kim, Ki-Young;Yi, Tae Hoo
    • Journal of Ginseng Research
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    • v.40 no.3
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    • pp.260-268
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    • 2016
  • Background: During the aging process, skin shows visible changes, characterized by a loss of elasticity and the appearance of wrinkles due to reduced collagen production and decreased elasticity of elastin fibers. Panax ginseng Meyer has been used as a traditional medicine for various diseases due to its wide range of biological activities including skin protective effects. Ginsenosides are the main components responsible for the biological activities of ginseng. However, the protective activities of an enzymatic preparation of red ginseng against human skin aging have not been investigated. Methods: The efficacy of an enzyme-treated powder complex of red ginseng (BG11001) in preventing human skin aging was evaluated by oral administration to 78 randomized individuals. All patients were requested to take three daily capsules containing either 750 mg of BG11001 or a placebo vehicle for 24 wk; at the end of the testing period, skin roughness, elasticity, and skin water content were measured. Results: BG11001 significantly reduced the average roughness of eye wrinkles and the Global Photo Damage Score compared with the placebo, although there were no significant differences in arithmetic roughness average between the groups. In addition, gross elasticity and net elasticity values increased, and transepidermal water loss level decreased, indicating improved skin elasticity and moisture content. Conclusion: In conclusion, enzyme-treated red ginseng extract significantly improved eye wrinkle roughness, skin elasticity, and moisture content. Moreover, enzyme-treated red ginseng extract would be useful substance as a bio-health skin care product.

Rice 7-Hydroxymethyl Chlorophyll a Reductase Is Involved in the Promotion of Chlorophyll Degradation and Modulates Cell Death Signaling

  • Piao, Weilan;Han, Su-Hyun;Sakuraba, Yasuhito;Paek, Nam-Chon
    • Molecules and Cells
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    • v.40 no.10
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    • pp.773-786
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    • 2017
  • The loss of green coloration via chlorophyll (Chl) degradation typically occurs during leaf senescence. To date, many Chl catabolic enzymes have been identified and shown to interact with light harvesting complex II to form a Chl degradation complex in senescing chloroplasts; this complex might metabolically channel phototoxic Chl catabolic intermediates to prevent oxidative damage to cells. The Chl catabolic enzyme 7-hydroxymethyl Chl a reductase (HCAR) converts 7-hydroxymethyl Chl a (7-HMC a) to Chl a. The rice (Oryza sativa) genome contains a single HCAR homolog (OsHCAR), but its exact role remains unknown. Here, we show that an oshcar knockout mutant exhibits persistent green leaves during both dark-induced and natural senescence, and accumulates 7-HMC a and pheophorbide a (Pheo a) in green leaf blades. Interestingly, both rice and Arabidopsis hcar mutants exhibit severe cell death at the vegetative stage; this cell death largely occurs in a light intensity-dependent manner. In addition, 7-HMC a treatment led to the generation of singlet oxygen ($^1O_2$) in Arabidopsis and rice protoplasts in the light. Under herbicide-induced oxidative stress conditions, leaf necrosis was more severe in hcar plants than in wild type, and HCAR-overexpressing plants were more tolerant to reactive oxygen species than wild type. Therefore, in addition to functioning in the conversion of 7-HMC a to Chl a in senescent leaves, HCAR may play a critical role in protecting plants from high light-induced damage by preventing the accumulation of 7-HMC a and Pheo a in developing and mature leaves at the vegetative stage.

Isolation and Characterization of Endocellulase-Free Multienzyme Complex from Newly Isolated Thermoanaerobacterium thermosaccharolyticum Strain NOI-1

  • Chimtong, Suphavadee;Tachaapaikoon, Chakrit;Pason, Patthra;Kyu, Khin Lay;Kosugi, Akihiko;Mori, Yutaka;Ratanakhanokchai, Khanok
    • Journal of Microbiology and Biotechnology
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    • v.21 no.3
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    • pp.284-292
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    • 2011
  • An endocellulase-free multienzyme complex was produced by a thermophilic anaerobic bacterium, Thermoanaerobacterium thermosaccharolyticum strain NOI-1, when grown on xylan. The temperature and pH optima for growth were $60^{\circ}C$ and 6.0, respectively. The bacterial cells were found to adhere to insoluble xylan and Avicel. A scanning electron microscopy analysis showed the adhesion of xylan to the cells. An endocellulase-free multienzyme complex was isolated from the crude enzyme of strain NOI-1 by affinity purification on cellulose and Sephacryl S-300 gel filtration. The molecular mass of the multienzyme complex was estimated to be about 1,200 kDa. The multienzyme complex showed one protein on native PAGE, one xylanase on a native zymogram, 21 proteins on SDS-PAGE, and 5 xylanases on a SDS zymogram. The multienzyme complex consisted of xylanase, ${\beta}$-xylosidase, ${\alpha}$-L-arabinofuranosidase, ${\beta}$-glucosidase, and cellobiohydrolase. The multienzyme complex was effective in hydrolyzing xylan and corn hulls. This is the first report of an endocellulase-free multienzyme complex produced by a thermophilic anaerobic bacterium, T. thermosaccharolyticum strain NOI-1.

NMR peak assignment for the elucidation of the solution structure of T4 Endonuclease V

  • Im, Hoo-Kang;Hyungmi Lihm;Yu, Jun-Suk;Lee, Bong-Jin
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1996.04a
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    • pp.183-183
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    • 1996
  • Bacteriophage T4 endonuclease V initiates the repair of ultraviolet (UV)-induced pyrimidine dimer photoproducts in duplex DNA. The mechanism of DNA strand cleavage involves four sequential stens: linear diffusion along dsDNA, pyrimidine dimer-specific binding,l pyrimidine dimer-DNA glycosylase activity, and Af lyase activity. Although crystal structure is known for this enzyme, solution structure has not been yet known. In order to elucidate the solution structure of this enzyme NMR spectroscopy was used. As a basis for the NMR peak assignment of the protein, HSQC spectrum was obtained on the uniformly $\^$15/N-labeled T4 endonuclease V. Each amide peak of the spectrum were classified according to amino acid spin systems by interpreting the spectrum of $\^$15/N amino acid-specific labeled T4 endonuclease V. The assignment was mainly obtained from three-dimensional NMR spectra such as 3D NOESY-HMQC, 3D TOCSY-HMQC. These experiments were carried out will uniformly $\^$15/N-labeled sample. In order to assign tile resonance of backbon atom, triple-resonance theree-dimensional NMR experiments were also performed using double labeled($\^$15/N$\^$13/C) sample. 3D HNCA, HN(CO)CA, HNCO, HN(CA)HA spectra were recorded for this purpose. The results of assignments were used to interpret the interaction of this enzyme with DNA. HSQC spectrum was obtained for T4 endonuclease V with specific $\^$15/N-labeled amino acids that have been known for important residue in catalysis. By comparing the spectrum of enzyme*DNA complex with that of the enzyme, we could confirm the important role of some residues of Thr, Arg, Tyr in activity. The results of assignments were also used to predict the secondary structure by chemical shift index (CSI).

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Studies on the Characterization of Cellulase Produced by Trichoderma viride QM 9414 (Trichoderma viride QM 9414가 생산하는 Cellulase 특성에 관한 연구)

  • 윤은숙;이혜정
    • The Korean Journal of Food And Nutrition
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    • v.3 no.1
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    • pp.57-68
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    • 1990
  • In order to obtain the fundamental informations on cellulase of Trichoderma viride QM 9414 for its production and utilization, some physico-chemical properties of the enzyme were reviewed. When T. viride QM 9414 was cultured on wheat bran medium, filter paper-disintegrating and carboxymethyl cellulose-saccharifying activity were increased with the cell growth, and thereafter CMC-saccharifying activity was kept on almost the same leved while filter-paper disintegrating activity was decreased sharply. And B-glucosidase was formed maximally on the late stationary phase of growth. The crude cellulase of cell-free extracts was purified by (NH4)2SO4 fractionation, Sephadex-G 200 column chromatography and DEAE Sephadex A-50 column chromatography. Filter paper-disintegrating, CMC-saccharifying and B-glucosidase activity were purified 10-fold, 47-fold and 38-fold, respectively. The crude enzyme was proved to be a complex of three different enzyme proteins which were showing filter paper-disintegrating, CMC-saccharifying and B-glucosidase activity. The optimal pH of the three enzyme components was alike pH 4.0, and the optimal temperature for CMC-saccharifying, filter paper-disintegrating and B-glucosidase activity were 4$0^{\circ}C$, 45$^{\circ}C$ and 5$0^{\circ}C$ respectively. The Km and Vmax values of CMC saccharifying activity for CMC were 0.485% and 3.10, and the Km and Vmax vallues of B-glucosidase for PNPG were 0.944$\times$10-3M and 0.097, respectively. The Km and Vmax values of filter paper-disintegrating activity for Avicel were determined to be 0.09% and 0.178, respectively. B-Glucosidase activity was competitively inhibited by glucose, and the Ki value for this enzyme was 3.54$\times$10-3M, CMC saccharifying activity was found to be greatly inhibited by cellobiose.

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Extracellular Wood-degradative Enzymes from Lentinus edodes JA01 (표고 균주(菌株) JA01에서 분비되는 세포외(細胞外) 목재성분(木材成分) 분해효소(分解酵素)에 관하여)

  • Hong, Soon-Woo;Shin, Kwang-Soo;Yoon, Yeup;Lee, Won-Kyu
    • The Korean Journal of Mycology
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    • v.14 no.3
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    • pp.189-194
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    • 1986
  • The aim of this study was to investigate physiological chracteristics of Lentinus edodes in Korea. We tried to detect properties of the several wood-degradative enzymes and investigate patterns of the enzyme production. A specific carbon source was used in the enzyme induction media for each enzymes, and the crude extract was used for the enzyme solution. With these enzyme solution, we investigated optimum temperature and pH conditions of their reactions. Moreover we investigated transition patterns of the enzyme production of the several wood-degrad­ative enzmes from Complex and Saw dust media for the purpose of studying the mechanisms of the wood component degradation by this fungus. It was assumed that the order of the wood com­ponent degradation was cellulose, xylan, and then pectic substances, and that the synergistic effects of these substances also influenced the degradation of wood components.

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