• Title/Summary/Keyword: rGH

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Novel Alkali-Tolerant GH10 Endo-${\beta}$-1,4-Xylanase with Broad Substrate Specificity from Microbacterium trichothecenolyticum HY-17, a Gut Bacterium of the Mole Cricket Gryllotalpa orientalis

  • Kim, Do Young;Shin, Dong-Ha;Jung, Sora;Kim, Hyangmi;Lee, Jong Suk;Cho, Han-Young;Bae, Kyung Sook;Sung, Chang-Keun;Rhee, Young Ha;Son, Kwang-Hee;Park, Ho-Yong
    • Journal of Microbiology and Biotechnology
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    • v.24 no.7
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    • pp.943-953
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    • 2014
  • The XylH gene (1,167-bp) encoding a novel hemicellulase (41,584 Da) was identified from the genome of Microbacterium trichothecenolyticum HY-17, a gastrointestinal bacterium of Gryllotalpa orientalis. The enzyme consisted of a single catalytic domain, which is 74% identical to that of an endo-${\beta}$-1,4-xylanase (GH10) from Isoptericola variabilis 225. Unlike other endo-${\beta}$-1,4-xylanases from invertebrate-symbiotic bacteria, rXylH was an alkali-tolerant multifunctional enzyme possessing endo-${\beta}$-1,4-xylanase activity together with ${\beta}$-1,3/${\beta}$-1,4-glucanase activity, which exhibited its highest xylanolytic activity at pH 9.0 and 60oC, and was relatively stable within a broad pH range of 5.0-10.0. The susceptibilities of different xylosebased polysaccharides to the XylH were assessed to be as follows: oat spelts xylan > beechwood xylan > birchwood xylan > wheat arabinoxylan. rXylH was also able to readily cleave p-nitrophenyl (pNP) cellobioside and pNP-xylopyranoside, but did not hydrolyze other pNP-sugar derivatives, xylobiose, or hexose-based materials. Enzymatic hydrolysis of birchwood xylan resulted in the product composition of xylobiose (71.2%) and xylotriose (28.8%) as end products.

The Production and Geochemistry of Evaporite from the Acid Mine Drainage (산성 광산배수로부터 형성되는 증발잔류광물의 생성량과 지구화학)

  • Park Cheon-Young;Cho Kap-Jin;Kim Seoung-Ku
    • Journal of the Korean earth science society
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    • v.26 no.6
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    • pp.524-540
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    • 2005
  • This study has focused on the amount of evaporites and geochemical characteritics of evaporites from the acid mine drainage and on the variation of constituents in acid mine drainage during evaporation. The various colors of evaporites are frequently observed at the rock surfaces contacting acid mine drainage. In order to produce evaporites in the laboratory, acid mine drainages were sampled from the abandoned mine areas (GTa, GTb, GH and GB) and air-dried at room temperature. During the evaporation of acid mine drainages, TDS, EC values and the concentrations of major and minor ions increased, whereas ER and DO values decreased with time. The concentration of Fe increased gradually with evaporation time in the GTb and GB, whereas GH founded in one day but rapidly not detected in the other day after due to removal of Fe by formation-precipitation of amorphous Fe hydroxide. The amounts of the evaporites were produced in amounts of 4 g (GTa), 5 g (GB), 15 g (GH), and 24 g (GTb) from 4 liter of acid mine drainage after 80 days of the evaporation, respectively. In linear analysis from the products with the parameters which are the EC, TDS, salinity, ER, DO and pH contents in field, the determination coefficients were 0.98, 0.99, 0.98, 0.88, 0.89, and 0.25 respectively. If we measure the parameters in field, it would be easy to estimate the amount of evaporites in acid mine drainage. Gypsum and epsomite were identified in all of the evaporites by x-ray powder diffraction studies. Evaporite (GTb) was heated at 52, 65, 70, 95, 150, 250, and 350oC for one hour in electrical furnaces. Gypsum, $CaSO_4\cdot1/2H_2O$ and kieserite were identified in the heated evaporite by XRD. With increased heating temperature, the intensity of the peak at $7.66/AA$ (diagnostic peak of gypsum), the peak at 5.59A ($CaSO_4{\cdot}1/2H_2O)$ and the peak at $4.83{\AA}$ (kieserite) decreased in x-ray diffraction due to dehydration. In the SEM and EDS analysis for the evaporite, gypsum of well-crystallized, radiating cluster of fibrous, acicular, and columnar shapes were observed in all samples. Ca was not detected in the EDS analysis of the flower structures of GTb. Because of that, the evaporite with flower structures is thought to be eposmite.

Cloning, Expression, and Characterization of a Thermostable GH51 ${\alpha}-\small{L}$-Arabinofuranosidase from Paenibacillus sp. DG-22

  • Lee, Sun Hwa;Lee, Yong-Eok
    • Journal of Microbiology and Biotechnology
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    • v.24 no.2
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    • pp.236-244
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    • 2014
  • The gene encoding ${\alpha}-\small{L}$-arabinofuranosidase (AFase) from Paenibacillus sp. DG-22 was cloned, sequenced, and expressed in Escherichia coli. The AFase gene (abfA) comprises a 1,509 bp open reading frame encoding 502 amino acids with a molecular mass of 56,520 daltons. The deduced amino acid sequence of the gene shows that AbfA is an enzyme consisting of only a catalytic domain, and that the enzyme has significant similarity to AFases classified into the family 51 of the glycosyl hydrolases. abfA was subcloned into the pQE60 expression vector to fuse it with a six-histidine tag and the recombinant AFase (rAbfA) was purified to homogeneity. The specific activity of the recombinant enzyme was 96.7 U/mg protein. Determination of the apparent molecular mass by gel-filtration chromatography indicated that AbfA has a tetrameric structure. The optimal pH and temperature of the enzyme were 6.0 and $60^{\circ}C$, respectively. The enzyme activity was completely inhibited by 1 mM $HgCl_2$. rAbfA was active only towards p-nitrophephenyl ${\alpha}-\small{L}$-arabinofuranoside and exhibited $K_m$ and $V_{max}$ values of 3.5 mM and 306.1 U/mg, respectively. rAbfA showed a synergistic effect in combination with endoxylanase on the degradation of oat spelt xylan and wheat arabinoxylan.

LC-MS/MS Method for Simultaneous Analysis of Growth Hormone-Releasing Peptides and Secretagogues in Human Urine

  • Min, Hophil;Han, Boyoung;Sung, Changmin;Park, Ju-Hyung;Lee, Kang Mi;Kim, Ho Jun;Kim, Ki Hun;Son, Junghyun;Kwon, Oh-Seung;Lee, Jaeick
    • Mass Spectrometry Letters
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    • v.7 no.3
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    • pp.55-63
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    • 2016
  • Growth hormone (GH)-releasing peptides (GHRPs) and GH secretagogues (GHSs) are listed in the World Anti-Doping Agency (WADA) Prohibited List. In the present study, we developed and validated a method for the simultaneous analysis of seven GHRPs (alexamorelin, GHRP-1, -2, -4, -5, -6, and hexarelin) and three GHSs (anamorelin, ibutamoren, and ipamorelin) in human urine. Method validation was performed at minimum required performance levels specified by WADA technical documents (2 ng/mL) for all substances, and the method was validated with regard to selectivity (no interference), linearity (R2 > 0.9986), matrix effects (50.0%-141.2%), recovery (10.4%-100.8%), and intra- (2.8%-16.5%) and inter-day (7.0%-22.6%) precisions. The limits of detection for screening and confirmation were 0.05-0.5 ng/mL and 0.05-1 ng/mL, respectively.

Optimization of a Medium for the Production of Cellulase by Bacillus subtilis NC1 Using Response Surface Methodology (반응 표면 분석법을 사용한 Bacillus subtilis NC1 유래 cellulase 생산 배지 최적화)

  • Yang, Hee-Jong;Park, Chang-Su;Yang, Ho-Yeon;Jeong, Su-Ji;Jeong, Seong-Yeop;Jeong, Do-Youn;Kang, Dae-Ook;Moon, Ja-Young;Choi, Nack-Shick
    • Journal of Life Science
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    • v.25 no.6
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    • pp.680-685
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    • 2015
  • Previously, cellulase and xylanase producing microorganism, Bacillus subtilis NC1, was isolated from soil. Based on the 16S rRNA gene sequence and API 50 CHL test the strain was identified as Bacillus subtilis, and named as B. subtilis NC1. We cloned and sequenced the genes for cellulase and xylanase. Plus, the deduced amino acid sequences from the genes of cellulase and xylanase were determined and were also identified as glycosyl hydrolases family (GH) 5 and 30, respectively. In this study to optimize the medium parameters for cellulase production by B. subtilis NC1 the RSM (response surface methodology) based on CCD (central composite design) model was performed. Three factors, tryptone, yeast extract, and NaCl, for N or C source were investigated. The cellulase activity was measured with a carboxylmethyl cellulose (CMC) plate and the 3,5-dinitrosalicylic acid (DNS) methods. The coefficient of determination (R2) for the model was 0.960, and the probability value (p=0.0001) of the regression model was highly significant. Based on the RSM, the optimum conditions for cellulase production by B. subtilis NC1 were predicted to be tryptone of 2.5%, yeast extract of 0.5%, and NaCl of 1.0%. Through the model verification, cellulase activity of Bacillus subtilis NC1 increased from 0.5 to 0.62 U/ml (24%) compared to the original medium.

Load analysis of an offshore monopile wind turbine using fully-coupled simulation (Fully-coupled 시뮬레이션을 이용한 해상 monopile 풍력 발전기의 응력해석)

  • Shi, Wei;Park, Hyun-Chul
    • 한국신재생에너지학회:학술대회논문집
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    • 2009.06a
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    • pp.480-485
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    • 2009
  • Offshore wind energy is gaining more attention. Ensuring proper design of offshore wind turbines and wind farms require knowledge of the external conditions in which the turbines and associated facilities are to operate. In this work, a three-bladed 5MW upwind wind turbine, which is supported by the monopile foundation, is studied by use of fully coupled aero-hydro-servo-elastic commercial simulation tool, 'GH-Bladed'$^{(R)}$. Specification of the structures are chosen from the OC3 (Offshore Code Comparison Collaboration) under "IEA Wind Annex XXIII-subtask2". The primary external conditions due to wind and waves are simulated. Design Load case 5.2 is investigated in this work. The steady state power curve and power production loads are evaluated. Comparison between different codes is made.

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An anomalous dissociation of protonated cluster ions of DNA guanine-cytosine base-pair

  • Seong, Yeon-Mi;Han, Sang-Yun;Jo, Sung-Chan;Oh, Han-Bin
    • Mass Spectrometry Letters
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    • v.2 no.3
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    • pp.73-75
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    • 2011
  • In the collisionally-activated dissociation of the proton-bound cluster ions of DNA base guanine (G) and cytosine (C), $G{\bullet}{\bullet}H^+{\bullet}{\bullet}C$, the abundance of [$CH^+$] ions was found to be higher than that of [$GH^+$] despite the fact that G has a higher proton affinity than C. This unexpected observation seems to demonstrate another example that the simple kinetic method scheme does not work. We suggest that a kinetic factor or detailed dynamics governing the proton transfer and dissociation should be carefully considered in the applications of the kinetic method to the proton affinity measurements.

Anti-idiotypic Antibodies against Bovine Growth Hormone

  • Verma, N.K.;Sodhi, R.;Rajput, Y.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.5
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    • pp.732-737
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    • 2003
  • Anti-antibodies against three mouse monoclonal antibodies viz. IIB5D6, VIA6E8 and VIC1F9 (specific to bovine growth hormone) in rabbits have been generated and characterized. Ammonium sulfate fractionated and affinity-purified monoclonal antibodies were used for producing anti-antibodies. The generated anti-antibodies were against common as well as uncommon antigenic determinants present in mouse monoclonal antibodies. The raised anti-antibodies replaced [$I^125$ ]bGH bound to goat liver microsomes indicating production of anti-idiotypic antibodies against bovine growth hormone. These antibodies can have profound implications in vivo in lactating bovines for enhancing milk yield.

INDUCTION OF GROWTH HORMONE RELEASE BY GLYCYRRHIZAE RADIX

  • Jung, Dae-Young;Lee, Ho-Young;Ha, Hye-Kyung;Jung, Da-Young;Yang, Ha-Ru;Lee, Je-Hyun;Kang, Sam-Sik;Kim, Chung-Sook
    • Proceedings of the PSK Conference
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    • 2003.04a
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    • pp.274.2-275
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    • 2003
  • The aim of this study was designed to determine the induction of rat growth hormone(rGH) by extracts of a popular herb, Glycyrrhizae radix(GR), roots of Glycyrrhiza glabra $\sub$Linne/, and Glycyrrhiza uralensis $\sub$Fischer/. In vitro study was carried out using primary rat pituitary cell culture for 3 days and then was treated with methanol extract corresponding to 1 mg of dried weight of herb per 1 $m\ell$ of culture solution. (omitted)

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Constraints on Cosmological Models from the Large-Scale Velocity Field

  • Doh, Jean-Gyung;Park, Changbom-;Chun, Mun-Suk
    • Bulletin of the Korean Space Science Society
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    • 1993.04a
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    • pp.16-16
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    • 1993
  • The Cosmic Mach number M is the ratio of the bulk flow velocity of the galaxrvelocity field on some scale R to the unall scale velocity dispersion within refcions of scale R. Because M is the ratio of two velocities, it is inn-dimansionat and the Here, independent of the amplitude of the power specHim and of the biasplnmeter in the linear theory. We have measured the Mach rnlmber for two observational samples: a spiral galaxy sample(AHM) of Aaronson and hiscoBlaborators with absolute distances measured by the infrared Ttillr-Fisher relatioa and an elliptical galaxy sample(EGALS) of Faber or 0, with distances determined by the relation. The effective depths distances of galaxies from the Local Group of these samples are 1639 km/s and 2862 e/s, respectivelr. The Machnumbers from these observed peculiar velocity Selds He fund as M=0.95 for AHMand M=0.59 for EGALS. We comPBre these calculated Mach numbers with thosefrom meck surweys drawn fuom three cosnulogical medels: the stand8rd biased nh=0.5 CDM modet an open CDM rrudel with gh=0.2, and a medd with thepower-law power specelm P(k)-k-1 and n=1. The Mach rnlmber test can give robust constraints on these cosmelogical nudels whose power spectra have very different shapes at large scales.

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