• Title/Summary/Keyword: endo-(1

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Isolation and Characterization of Marine Bacterial Strain Degrading Fucoidan from Korean Undaria pinnatifida Sporophylls

  • Kim, Woo-Jung;Kim, Sung-Min;Lee, Yoon-Hee;Kim, Hyun-Guell;Kim, Hyung-Kwon;Moon, Seong-Hoon;Suh, Hyun-Hyo;Jang, Ki-Hyo;Park, Yong-Il
    • Journal of Microbiology and Biotechnology
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    • v.18 no.4
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    • pp.616-623
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    • 2008
  • In spite of an increasing interest in fucoidans as biologically active compounds, no convenient commercial sources with fucoidanase activity are yet available. A marine bacterial strain that showed confluent growth on a minimal medium containing fucoidan, prepared from Korean Undaria pinnatifida sporophylls, as the sole carbon source was isolated and identified based on a 16S rDNA sequence analysis as a strain of Sphingomonas paucimobilis, and named Sphingomonas paucimobilis PF-1. The strain depolymerized fucoidan into more than 7 distinct low-molecular-mass fucose-containing oligosaccharides, ranging from 305 to 3,749 Da. The enzyme activity was shown to be associated with the whole cell, suggesting the possibility of a surface display of the enzyme. However, a whole-cell enzyme preparation neither released the monomer L-fucose from the fucoidan nor hydrolyzed the chromogenic substrate p-nitrophenyl-${\alpha}$-L-fucoside, indicating that the enzyme may be an endo-acting fucoidanase rather than an ${\alpha}$-L-fucosidase. Therefore, this would appear to be the first report on fucoidanolytic activity by a Sphingomonas species and also the first report on the enzymatic degradation of the Korean Undaria pinnatifida sporophyll fucoidan. Moreover, this enzyme activity may be very useful for structural analyses of fucose-containing polysaccharides and the production of bioactive fucooligosaccharides.

Screening Differential Expressions of Defense-related Responses in Cold-treated 'Kyoho' and 'Campbell Early' Grapevines

  • Ahn, Soon Young;Kim, Seon Ae;Han, Jae Hyun;Kim, Seung Heui;Yun, Hae Keun
    • Horticultural Science & Technology
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    • v.31 no.3
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    • pp.275-281
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    • 2013
  • Low temperature is one of the major environmental factors that affect productivity including reduced growth and budding of vines, and changes of metabolic processes in grape (Vitis spp.). To screen the specific expression of abiotic stress-related genes against cold treatment in 'Kyoho' and 'Campbell Early' grapevines, expression of various defense-related genes was investigated by RT-PCR and real-time PCR. Among the 67 genes analyzed by RT-PCR and real-time PCR, 17 and 16 types of cDNA were up-regulated, while 5 and 6 types were down-regulated in cold-treated 'Kyoho' and 'Campbell Early' grapevines, respectively. Genes encoding carotene (Cart3564 and Cart4472), chalcone isomerase (CHI), cytochrome P450 (CYP), flavonol synthase (FLS), endo-${\beta}$-glucanase precursor (Glu), glutathione peroxidase (GPX), glutathione-S-transferase (GST), leucine-rich repeats (LRR), manganese superoxide dismutase (Mn-SOD), phenylalanine ammonia lyase (PAL), polygalacturonase-inhibiting protein (PGIP), proline rich protein 2 (PRP2), small heat shock protein (sHSP), temperature induced lipocalin (TIL), and thaumatin-like protein (TLP) were up-regulated, while those encoding CBF like transcription factor (CBF1), chitinase-like protein (CLP), cold induced protein (CIP), glycerol-3-phosphate acyltransferase (GPAT), and mitogen-activated protein kinase (MAPK) were down-regulated by low temperature treatment in both in 'Kyoho' and 'Campbell Early'.

Modification of Pullulan Using Dextransucrase and Characterization of the Modified Pullulan. (덱스트란수크라제를 이용한 플루란의 변형 및 특성조사)

  • ;;;;;;John F. Robyt
    • Microbiology and Biotechnology Letters
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    • v.26 no.3
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    • pp.264-268
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    • 1998
  • Many enzymes catalyze a primary reaction and/or secondary reaction. Dextransucrase usually synthesizes dextran from sucrose as a primary reaction. The secondary reaction of dextransucrase is the transfer of glucose from sucrose to carbohydrate accepters. We have reacted dextransucrase from Leuconostoc mesenteroides B-742CB with sucrose and pullulan as an acceptor under different reaction conditions; various concentrations of pullulan, enzyme, sucrose and different pHs and temperatures of reaction digests. The yield of modified pullulan was 57%(<${\pm}$5%) of theoretical under the reaction condition of pH 5.2, temperature 28$^{\circ}C$, 0.37% of pullulan, and 0.l U/$m\ell$ of dextransucrase. Modified products were more resistant against the hydrolysis of pullulanase and endo-dextranase than those of native pullulan. The positions of glucose substitution in the modified products were determined by methylation followed by acid hydrolysis and analyzed by TLC. The products were modified by the addition of glucose to the position of C3, C4, C6 free hydroxyl group of glucose residues in the pullulan.

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Purification and Characterization of Carboxymethyl Cellulase IV from Penicillium verruculosum (Penicillium verruculosum 으로부터 Carboxymethyl Cellulase IV 의 정제(精製) 및 특성(特性))

  • Kim, Jeong-Ho;Lee, Jae-Chang;Lee, Yong-Kyu;Kim, Kang-Hwa;Chun, Soon-Bai;Chung, Ki-Chul
    • The Korean Journal of Mycology
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    • v.21 no.1
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    • pp.28-37
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    • 1993
  • An endo-type cellulase, carboxymethyl cellulase(CMCase) IV, was purified from culture filtrate of cellulolytic fungus Penicillium verruculosum. The CMCase IV was acidic glycoprotein having carbohydrate of 13% as glucose and pI value of 4.0. The CMCase IV was 52 KDa of molecular weight in SDS-polyacrylamide gel electrophoresis and have optimum temperature and pH of $50^{\circ}C$ and 5.0 for enzyme activity. The CMCase IV liberated glucose and cellobiose as major products of the enzyme against carboxymethyl cellulose(CMC) and seemed to has transglycosylation activity simultaneously. Cellulase activity staining(zymogram) showed that the cellulase components of P. verruculosum were not aggregated in the medium. P. vrttuculosum mRNA was translated in vitro and translation product by the mRNA coding for CMCase IV was identified by immunoprecipitation.

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Endochitosanase Produced by Bacillus sp. P2l as a Potential Source for the Production of Chitooligosaccharides. (키토산 올리고당의 제조용 소재로서 Bacillus sp. P2l 기원의 키토산분해효소)

  • 박노동;조유영;이현철;조종수;조도현
    • Microbiology and Biotechnology Letters
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    • v.26 no.4
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    • pp.345-351
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    • 1998
  • In an effort to develop a potent system for the production of various dp (degree of polymerization) chitooligosaccharides, 32 enzymes or microbial systems were screened for chitosanolytic acitivity using chitosan as a substrate. The efficiency of each enzyme system was evaluated by the changes of turbidity and viscosity of chitosan solution, the amount of precipitate and the reducing sugar-producing activity in the enzymatic reaction mixture. Based on these assay methods for the chitosanase activity, Bacillus sp. P2l out of 32 screened systems showed highly potent endochitosanase, which was comparable with a commercially available enzyme (E7). Chitooligosaccharides of dp 3-7 were separated by TLC as major enzymatic reaction products, suggesting that the chitosanase from Bacillus sp. P2l be endo-splitting type.

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Comparison of Solid Waste Stabilization and Methane Emission from Anaerobic and Semi-Aerobic Landfills Operated in Tropical Condition

  • Sutthasil, Noppharit;Chiemchaisri, Chart;Chiemchaisri, Wilai;Wangyao, Komsilp;Towprayoon, Sirintornthep;Endo, Kazuto;Yamada, Masato
    • Environmental Engineering Research
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    • v.19 no.3
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    • pp.261-268
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    • 2014
  • Leachate quality and methane emission from pilot-scale lysimeters operated under semi-aerobic and anaerobic conditions were monitored for 650 days. Two semi-aerobic lysimeters were filled with un-compacted and compacted municipal solid wastes whereas two anaerobic lysimeters containing compacted wastes were operated with leachate storage at 50% and 100% of waste height, respectively. Despite having high moisture in wastes and operating under tropical rainfall events, leachate stabilization in semi-aerobic lysimeters took place much faster resulting in BOD reduction by 90% within 60 days, significantly shorter than 180-210 days observed in anaerobic lysimeters. Nitrogen concentration in leachate from semi-aerobic lysimeter could be reduced by 90%. In term of gas emission, semi-aerobic lysimeter with un-compacted wastes had much lower methane emission rate of $2.8g/m^2/day$ compare to anaerobic lysimeters ($62.6g/m^2/day$) through seasonal fluctuation was observed. Nevertheless, semi-aerobic lysimeter with waste compaction has similar performance to anaerobic lysimeter.

Effect of internal stress on cyclic fatigue failure in .06 taper ProFile (내부 응력이 .06 taper ProFile의 피로 파절에 미치는 영향)

  • Jung, Hye-Rim;Kim, Jin-Woo;Cho, Kyung-Mo;Park, Se-Hee
    • Restorative Dentistry and Endodontics
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    • v.37 no.2
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    • pp.79-83
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    • 2012
  • Objectives: The purpose of this study was to evaluate the relation between intentionally induced internal stress and cyclic fatigue failure of .06 taper ProFile. Materials and Methods: Length 25 mm, .06 taper ProFile (Dentsply Maillefer), and size 20, 25, 30, 35 and 40 were used in this study. To give the internal stress, the rotary NiTi files were put into the .02 taper, Endo-Training-Bloc (Dentsply Maillefer) until auto-stop by torque controlled motor. Rotary NiTi files were grouped by the number of induced internal stress and randomly distributed among one control group and three experimental groups (n = 10, Stress 0 [control], Stress 1, Stress 2 and Stress 3). For cyclic fatigue measurement, time for separation of the rotary NiTi files was recorded. The fractured surfaces were observed by field emission scanning electron microscope (FE-SEM, SU-70, Hitachi). The time for separation was statistically analyzed using two-way ANOVA and post-hoc Scheffe test at 95% level. Results: In .06 taper ProFile size 20, 25, 30, 35 and 40, there were statistically significant difference on time for separation between control group and the other groups (p < 0.05). Conclusion: In the limitation of this study, cyclic fatigue failure of .06 taper ProFile is influenced by internal stress accumulated in the files.

Characterization of Carboxymethylcellulase(CMCase) Produced by Recombinant E. coli Containing CMCase Gene for Cellulomonas sp. YE-5

  • Park, Sung-Won;Her, Nam-Yun;Kim, Dong-Seob;Park, Sun-Jin;Lee, Han-Seung;Park, Hak-Jong;Yu, Ju-Hyun
    • Preventive Nutrition and Food Science
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    • v.2 no.2
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    • pp.174-179
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    • 1997
  • CMCase produced by recombinant E. coli JM109 (pCEH#4) containing CMCase gene from Cellulomonas sp. YE-5 was purified to 24.3 fold and 2.6% yield by ammoniumsulfate precipitation, DEAE-cellulose column chromatography and gel filtration on Sephadex G-100. The optimum pH and temperature for CMCase activity were pH 7.0 and 5$0^{\circ}C$. The enzyme was stable between pH 5.0 and 10.0, and up to 6$0^{\circ}C$. The molecular weight of he enzyme was estimated to be approximately 40,000 daltons by SDS-PAGE. Analysis of the amino acid composition showed that the enzyme contained many glycines and acidic amino acids. The enzyme was an endo-type CMCase and the final enzyme reaction product from hydrolysis of Cm-cellulose by the enzyme was cellobiose. {TEX}$K_{M}${/TEX} value determined with CM-cellulose was 1.28mM.

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Cyclic fatigue, bending resistance, and surface roughness of ProTaper Gold and EdgeEvolve files in canals with single- and double-curvature

  • Khalil, Wafaa A.;Natto, Zuhair S.
    • Restorative Dentistry and Endodontics
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    • v.44 no.2
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    • pp.19.1-19.9
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    • 2019
  • Objectives: The purpose of this study was to evaluate the cyclic fatigue, bending resistance, and surface roughness of EdgeEvolve (EdgeEndo) and ProTaper Gold (Dentsply Tulsa Dental Specialties) nickel-titanium (NiTi) rotary files. Materials and Methods: The instruments (n = 15/each) were tested for cyclic fatigue in single- ($60^{\circ}$ curvature, 5-mm radius) and double-curved (coronal curvature $60^{\circ}$, 5-mm radius, and apical curvature of $30^{\circ}$ and 2-mm radius) artificial canals. The number of cycles to fracture was calculated. The bending resistance of both files were tested using a universal testing machine where the files were bent until reach $45^{\circ}$. Scanning electron microscopy and x-ray energy-dispersive spectrometric analysis were used for imaging the fractured segments, while the atomic force microscope was used to quantify the surface roughness average (Ra). Results: EdgeEvolve files exhibited higher cyclic fatigue resistance than ProTaper Gold files in single- and double-curved canals (p < 0.05) and both files were more resistant to cyclic fatigue in single-curved canals than double-curved canals (p < 0.05). EdgeEvolve files exhibited significantly more flexibility than did ProTaper Gold files (p < 0.05). Both files had approximately similar Ni and Ti contents (p > 0.05). EdgeEvolve files showed significantly lower Ra values than ProTaper Gold files (p < 0.05). Conclusions: Within the limitation of this study, EdgeEvolve files exhibited significantly higher cyclic fatigue resistance than ProTaper Gold files in both single- and double-curved canals.

Effect of irrigation protocols on smear layer removal, bond strength and nanoleakage of fiber posts using a self-adhesive resin cement

  • Rodrigo Stadler Alessi;Renata Terumi Jitumori ;Bruna Fortes Bittencourt;Giovana Mongruel Gomes ;Joao Carlos Gomes
    • Restorative Dentistry and Endodontics
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    • v.48 no.3
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    • pp.28.1-28.13
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    • 2023
  • Objectives: This study aimed to investigate the effect of the application method of 2% chlorhexidine (CHX) and its influence on the adhesion of fiberglass posts cemented with a self-adhesive resin cement. Materials and Methods: Sixty human mandibular premolars were endodontically treated and divided into 5 groups (n = 12), according to the canal irrigant and its application method: 2 groups with conventional syringe irrigation (CSI)-2.5% sodium hypochlorite (NaOCl) (control) and 2% CHX- and 3 groups with 2% CHX irrigation/activation-by passive ultrasonic irrigation (PUI), Easy Clean file, and XP-Endo Finisher file. Two roots per group were evaluated for smear layer (SL) removal by scanning electron microscopy. For other roots, fiber posts were luted using a self-adhesive resin cement. The roots were sectioned into 6 slices for push-out bond strength (BS) (7/group) and nanoleakage (NL) (3/group). Data from SL removal were submitted to Kruskal-Wallis and Student-Newman-Keuls tests (α = 0.05). Data from BS and NL were evaluated by 2-way analysis of variance and Tukey's test (α = 0.05). Results: For SL removal and BS, the CHX irrigation/activation promoted better values than CSI with CHX (p < 0.05), but it was not significantly different from CSI with NaOCl (p > 0.05). For NL, the lowest values were obtained by the chlorhexidine irrigation/activation groups (p < 0.05). Conclusions: Active 2% CHX irrigation can be used to improve the post space cleaning and adhesion before fiber post cementation with self-adhesive resin cements.