• Title/Summary/Keyword: endo-(1

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Cloning, Expression, and Characterization of Protease-resistant Xylanase from Streptomyces fradiae var. k11

  • Li, Ning;Yang, Peilong;Wang, Yaru;Luo, Huiying;Meng, Kun;Wu, Nigfeng;Fan, Yunliu;Yao, Bin
    • Journal of Microbiology and Biotechnology
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    • v.18 no.3
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    • pp.410-416
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    • 2008
  • The gene SfXyn10, which encodes a protease-resistant xylanase, was isolated using colony PCR screening from a genomic library of a feather-degrading bacterial strain Streptomyces fradiae var. k11. The full-length gene consists of 1,437bp and encodes 479 amino acids, which includes 41 residues of a putative signal peptide at its N terminus. The amino acid sequence shares the highest similarity (80%) to the endo-1,4-${\beta}$-xylanase from Streptomyces coelicolor A3, which belongs to the glycoside hydrolase family 10. The gene fragment encoding the mature xylanase was expressed in Escherichia coli BL21 (DE3). The recombinant protein was purified to homogeneity by acetone precipitation and anion-exchange chromatography, and subsequently characterized. The optimal pH and temperature for the purified recombinant enzyme were 7.8 and $60^{\circ}C$, respectively. The enzyme showed stability over a pH range of 4.0-10.0. The kinetic values on oat spelt xylan and birchwood xylan substrates were also determined. The enzyme activity was enhanced by $Fe^{2+}$ and strongly inhibited by $Hg^{2+}$ and SDS. The enzyme also showed resistance to neutral and alkaline proteases. Therefore, these characteristics suggest that SfXyn10 could be an important candidate for protease-resistant mechanistic research and has potential applications in the food industry, cotton scouring, and improving animal nutrition.

Optimization of an Extracellular Dextranase Production from Lipomyces starkeyi KCTC 17343 and Analysis of Its Dextran Hydrolysates (Lipomyces starkeyi KCTC 17343에 의한 extracellular dextranase 최적생산과 덱스트란 hydrolysates 분석)

  • Chang, Yoon-Hyuck;Yeom, Joong-Hyun;Jung, Kyung-Hwan;Chang, Byung-Chul;Shin, Jung-Hee;Yoo, Sun-Kyun
    • Journal of Life Science
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    • v.19 no.4
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    • pp.457-461
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    • 2009
  • We optimized dextranase culture conditions by batch fermentation using Lipomyces starkeyi KCTC 17343. Furthermore, dextranase was purified by an ultra-membrane, and then dextran hydrolyzates were characterized. Cell growth and dextranase production varied depending on the initial culture pH and temperature. The conditions of optimal dextranase production were met in a pH range of 4-5 and temperature between $25-30^{\circ}C$. At optimal fermentation conditions, total enzyme activity and specific enzyme activity were about 4.85 IU/ml and 0.79 IU/g cells, respectively. The specific growth rate was examined to be $0.076\;hr^{-1}$. The production of dextranase in culture broth was very stably maintained after mid-log phase of growth. The enzyme hydrolyzed dextran into DP (degree of polymerization) 2 to 8 oligodextran series. Analysis of the composition of hydrolysates suggested that the enzyme produced is an endo-dextranase.

Changes in Pectin and Pectin Degrading Enzymes Activity during Storage of Kiyomi Tangor Produced in Jeju (제주산 만감류 청견의 저장 중 펙틴 및 펙틴분해효소 활성의 변화)

  • 강문장;임자훈;고정삼
    • Food Science and Preservation
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    • v.8 no.2
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    • pp.131-139
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    • 2001
  • Kiyomi tangor(Citus unshiu x sinensis) was stored at 3$\^{C}$ and 85% relative humidity, and the changes in firmness, pectin degrading enzymes activity and other physicochemical properties of citrus fruits during storage were investigated. Decay ratio and weight loss during 180 days’ storage were increased gradually to 13.0% and 12.9%, respectively. Firmness of fruits with 2 mm probe was decreased gradually from 808.7 g-force to 406.4 g-force, and moisture of peel and flesh were decreased from 76.5% to from 89.6% to 87.6% during storage, respectively. Exo-polygalacturonase activity of peel after 150 days’ storage were increased gradually to 558.09 units/100g. Pectin methylesterase activity of peel and flesh were increased from 14.7 units/g to 2.3 units/g, and from 9.4 units/ml to 2.7 units/ml at 150days’ storage, respectively. Endo-polygalacturonase activities were not changed notably during storage. Alcohol-insoluble solid(AIS) of peel was not changed notably. During storage of the fruits water soluble pectin(WSP) of peel and flesh were increased from 474.49 mg/100g to 614.29mg/100g, and from 66.91mg/100g to 92.74mg/100g as wet basis, respectively. Hexameta-phosphate soluble pectin(HMP) of peel were decreased from 405.5mg/100g to 270.43mg/100g, hydochloric acid soluble pectin(HSP) of peel was also decreased from 544.02mg/100g to 412.64mg/100g during storage. Total pectin substance(TPS) of peel and flesh were decreased from 1,424.01mg/100g to 1,297.36mg/100g, and from 165.51mg/100g to 171.54mg/100g, respectively. Composition ratio of pectin was in order of WSP > HSP > HMP.

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Screening and Characterization of a Novel Cellulase Gene from the Gut Microflora of Hermetia illucens Using Metagenomic Library

  • Lee, Chang-Muk;Lee, Young-Seok;Seo, So-Hyeon;Yoon, Sang-Hong;Kim, Soo-Jin;Hahn, Bum-Soo;Sim, Joon-Soo;Koo, Bon-Sung
    • Journal of Microbiology and Biotechnology
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    • v.24 no.9
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    • pp.1196-1206
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    • 2014
  • A metagenomic fosmid library was constructed using genomic DNA isolated from the gut microflora of Hermetia illucens, a black soldier fly. A cellulase-positive clone, with the CS10 gene, was identified by extensive Congo-red overlay screenings for cellulase activity from the fosmid library of 92,000 clones. The CS10 gene was composed of a 996 bp DNA sequence encoding the mature protein of 331 amino acids. The deduced amino acids of CS10 showed 72% sequence identity with the glycosyl hydrolase family 5 gene of Dysgonomonas mossii, displaying no significant sequence homology to already known cellulases. The purified CS10 protein presented a single band of cellulase activity with a molecular mass of approximately 40 kDa on the SDS-PAGE gel and zymogram. The purified CS10 protein exhibited optimal activity at $50^{\circ}C$ and pH 7.0, and the thermostability and pH stability of CS10 were preserved at the ranges of $20{\sim}50^{\circ}C$ and pH 4.0~10.0. CS10 exhibited little loss of cellulase activity against various chemical reagents such as 10% polar organic solvents, 1% non-ionic detergents, and 0.5 M denaturing agents. Moreover, the substrate specificity and the product patterns by thin-layer chromatography suggested that CS10 is an endo-${\beta}$-1,4-glucanase. From these biochemical properties of CS10, it is expected that the enzyme has the potential for application in industrial processes.

Changes in terpenes of three kinds of pine needles during litter decomposition

  • Jo, Gyu-Gap;Kim, Jong-Hee
    • Journal of Ecology and Environment
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    • v.33 no.2
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    • pp.175-186
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    • 2010
  • This study was designed to evaluate changes in the terpene composition of 3 types of pines (Pinus densiflora, Pinus thunbergii and Pinus rigida), while decomposing their leaf litter. Needle litters were placed at two different organic layer depths, one on the surface and the other beneath the litter layer. Changes in the terpene composition of this litter were detected using a gas chromatograph-mass spectrometer. Among the monoterpenes acquired from the fresh needles of P. densiflora and P. rigida, $\alpha$-pinene (12.05% and 19.87%, respectively) was the major one, followed by $\beta$-pinene (2.90% and 14.07%). However, from the needles of P. thunbergii, $\beta$-pinene (20.77%) was the major one, followed by $\alpha$-pinene (10.79%). Among the sesquiterpenes detected in P. densiflora, trans-caryophyllene (3.12%) was the highest composition compound, whereas germacrene-D (6.09%) for P. thunbergii and 1,6-cyclodecadiene (7.41%) and endo-1-bourbonanol (7.41%) for P. rigida were the highest content compounds. However, the total amounts of terpenes decreased sharply by 40-85.4% in all three types of pine needle after 90-120 days of the experiment. The concentration of each terpene differed during decomposition, and the majority of compounds disappeared from beneath the litter layer. It was determined that three types of reducing patterns of each compound appeared on the rate of loss of concentration during decomposition; one pattern decreasing sharply during the initial period, another pattern steadily or slowly decreasing, and a newly detected pattern at low concentration occurring during decomposition.

Development and Performance Evaluation of the First Model of 4D CT-Scanner

  • Endo, Masahiro;Mori, Shinichiro;Tsunoo, Takanori;Kandatsu, Susumu;Tanada, Shuji;Aradate, Hiroshi;Saito, Yasuo;Miyazaki, Hiroaki;Satoh, Kazumasa;Matsusita, Satoshi;Kusakabe, Masahiro
    • Proceedings of the Korean Society of Medical Physics Conference
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    • 2002.09a
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    • pp.373-375
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    • 2002
  • 4D CT is a dynamic volume imaging system of moving organs with an image quality comparable to conventional CT, and is realized with continuous and high-speed cone-beam CT. In order to realize 4D CT, we have developed a novel 2D detector on the basis of the present CT technology, and mounted it on the gantry frame of the state-of-the-art CT-scanner. In the present report we describe the design of the first model of 4D CT-scanner as well as the early results of performance test. The x-ray detector for the 4D CT-scanner is a discrete pixel detector in which pixel data are measured by an independent detector element. The numbers of elements are 912 (channels) ${\times}$ 256 (segments) and the element size is approximately 1mm ${\times}$ 1mm. Data sampling rate is 900views(frames)/sec, and dynamic range of A/D converter is 16bits. The rotation speed of the gantry is l.0sec/rotation. Data transfer system between rotating and stationary parts in the gantry consists of laser diode and photodiode pairs, and achieves net transfer speed of 5Gbps. Volume data of 512${\times}$512${\times}$256 voxels are reconstructed with FDK algorithm by parallel use of 128 microprocessors. Normal volunteers and several phantoms were scanned with the scanner to demonstrate high image quality.

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Expression of CYP2A6, CYP2D6 and CYP4A11 Polymorphisms in COS7 Mammalian Cell Line

  • Lee, Hye-Ja;Park, Mi-Kyung;Park, Young-Ran;Kim, Dong-Hak;Yun, Chul-Ho;Chun, Young-Jin;Shin, Hee-Jung;Na, Han-Sung;Chung, Myeon-Woo;Lee, Chang-Hoon
    • Toxicological Research
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    • v.27 no.1
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    • pp.25-29
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    • 2011
  • The cytochrome P450 (P450, CYP) are the superfamily of heme-containing monooxygenase enzymes, found throughout all nature including mammals, plants, and microorganisms. Mammalian P450 enzymes are involved in oxidative metabolism of a wide range of endo- and exogenous chemicals. Especially P450s involved in drug metabolisms are important for drug efficacy and polymorphisms of P450s in individuals reflect differences of drug responses between people. To study the functional differences of CYP2A6, CYP2D6, and CYP4A11 variants, we cloned the four CYP2A6, three CYP2D6, and three CYP4A11 variants, which were found in Korean populations, in mammalian expression vector pcDNA by PCR and examined their expressions in COS-7 mammalian cells using immunoblots using P450 specific polyclonal antibodies. Three of four CYP2A6, two of three CYP4A11, and two of three CYP2D6 variants showed expressions in COS-7 cells but the relative levels of expressions are remarkably different in those of each variants. Our findings may help to study and explain the differences between functions of CYP variants and drug responses in Korean populations.

Environmentally Friendly Preparation of Functional Nanomaterials and Their Application

  • Lee, Sun-Hyung;Teshima, Katsuya;Endo, Morinobu;Oishi, Shuji
    • Proceedings of the Materials Research Society of Korea Conference
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    • 2010.05a
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    • pp.5.1-5.1
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    • 2010
  • One of the most important environmental problems is global warming. Global warming is caused by increase in the amounts of water vapor, methane, carbon dioxide and other gases being released into the atmosphere as a result of the burning of fossil fuels. It has thus become important to reduce fossil fuel use. Environmentally friendly preparation of functional materials has, therefore, attracted much interest for environmental problems. Furthermore, nature mimetic processes are recently been of great interest as environmentally friendly one. There have been many studies on fabrication of various functional nanocrystals. Among various nanocrystal fabrication techniques, flux growth is an environmentally friendly, very convenient process and can produce functional nanocrystals at temperatures below the melting points of the solutes. Furthermore, this technique is suitable for the synthesis of crystals having an enhedral habit. In flux growth, the constituents of the materials to be crystallized are dissolved in a suitable flux (solvent) and crystal growth occurs as the solution becomes critically supersaturated. The supersaturation is attained by cooling the solution, by evaporation of the solvent or by a transport process in which the solute is made to flow from a hotter to a cooler region. Many kinds of oxide nanocrystals have been grown in our laboratory. For example, zero- (e.g., particle), one- (e.g., whisker and tube) and two-dimensional (e.g., sheet) nanocrystals were successfully grown by flux method. Our flux-growth technique has some industrial and ecological merits because the nanocrystal fabrication temperatures are far below their melting points and because the used reagents are less harmless to human being and the environment.

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Phylogenetics and Gene Structure Dynamics of Polygalacturonase Genes in Aspergillus and Neurospora crassa

  • Hong, Jin-Sung;Ryu, Ki-Hyun;Kwon, Soon-Jae;Kim, Jin-Won;Kim, Kwang-Soo;Park, Kyong-Cheul
    • The Plant Pathology Journal
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    • v.29 no.3
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    • pp.234-241
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    • 2013
  • Polygalacturonase (PG) gene is a typical gene family present in eukaryotes. Forty-nine PGs were mined from the genomes of Neurospora crassa and five Aspergillus species. The PGs were classified into 3 clades such as clade 1 for rhamno-PGs, clade 2 for exo-PGs and clade 3 for exo- and endo-PGs, which were further grouped into 13 sub-clades based on the polypeptide sequence similarity. In gene structure analysis, a total of 124 introns were present in 44 genes and five genes lacked introns to give an average of 2.5 introns per gene. Intron phase distribution was 64.5% for phase 0, 21.8% for phase 1, and 13.7% for phase 2, respectively. The introns varied in their sequences and their lengths ranged from 20 bp to 424 bp with an average of 65.9 bp, which is approximately half the size of introns in other fungal genes. There were 29 homologous intron blocks and 26 of those were sub-clade specific. Intron losses were counted in 18 introns in which no obvious phase preference for intron loss was observed. Eighteen introns were placed at novel positions, which is considerably higher than those of plant PGs. In an evolutionary sense both intron loss and gain must have taken place for shaping the current PGs in these fungi. Together with the small intron size, low conservation of homologous intron blocks and higher number of novel introns, PGs of fungal species seem to have recently undergone highly dynamic evolution.

An in vitro evaluation of the accuracy of four electronic apex locators using stainless-steel and nickel-titanium hand files

  • Gehlot, Paras Mull;Manjunath, Vinutha;Manjunath, Mysore Krishnaswamy
    • Restorative Dentistry and Endodontics
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    • v.41 no.1
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    • pp.6-11
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    • 2016
  • Objectives: The purpose of this in vitro study was to evaluate the accuracy of working length (WL) determination of four electronic apex locators (EALs), namely, Root ZX (RZX), Elements diagnostic unit and apex locator (ELE), SybronEndo Mini Apex locator (MINI) and Propex pixi (PIXI) using Stainless steel (SS) and nickel-titanium (NiTi) hand files. The null hypothesis was that there was no difference between canal length determination by SS and NiTi files of 4 EALs. Materials and Methods: Sixty extracted, single rooted human teeth were decoronated and the canal orifice flared. The actual length (AL) was assessed visually, and the teeth were embedded in an alginate model. The electronic length (EL) measurements were recorded with all four EALs using SS and NiTi files at '0.5' reading on display. The differences between the AL and EL were compared. Results: The results obtained with each EAL with SS and NiTi files were compared with AL. A paired sample t test showed that there was a statistical significant difference between EAL readings with SS and NiTi files for RZX and MINI (p < 0.05). The accuracy of RZX, ELE, MINI and PIXI within ${\pm}0.5 mm$ of AL with SS/NiTi files were 93.3%/70%, 90%/91.7%, 95%/68.3%, and 83.3%/83.3%, respectively. Conclusions: The results of this study indicate that Root ZX was statistically more accurate with NiTi files compared to SS files, while MINI was statistically more accurate with SS files compared to NiTi files. ELE and PIXI were not affected by the alloy type of the file used to determine WL.