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Domain Function and Relevant Enzyme Activity of Cycloinulooligosaccharide Fructanotransferase from Paenibacillus polymyxa (Paenibacillus polymyxa Cycloinulooligosaccharide Fructanotransferase의 효소 활성에 미치는 각 Domain의 역할)

  • You Dong-Ju;Park Jung-Ha;You Kyung-Ok;Nam Soo-Wan;Kim Kwang-Hyeon;Kim Byung-Woo;Kwon Hyun-Ju
    • Microbiology and Biotechnology Letters
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    • v.34 no.3
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    • pp.278-287
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    • 2006
  • Cycloinulooligosaccharide fructanotransferase (CFTase) converts inulin into cycloinulooligosaccharides (cyclofructan, CF) of ${\beta}-(2{\to}1)$-linked D-fructofuranose as well as hydrolysis of cyclofructan. Sequences analysis indicated that CFTase was divided into five distinct regions containing three repeated sequences (R1, R3, and R4) at the N-terminus and C-terminus. Each domain function was investigated by comparison of wild type CFTase enzyme (CFT148) and deletion mutant proteins (CFT108: R1 and R3 deletion; CFT130: R4 deletion; and CFT88: R1, R3, and R4 deletion) of CFTase. The CFT108 mutant had both CFTase and CF hydrolyzing activity as CFT148 did. CFTase activities and CF hydrolysing activities were disappeared in CFT130 and CFT88 mutants. These results indicated that the C-terminal R4 region of P. polymyxa CFTase is necessary for cyclization and hydrolyzing activity.

Purification of Two Novel Antimicrobial Peptides from Pyloric Caeca of the Starfish Asterina pectinifera (별불가사리 Asterina pectinifera의 유문맹낭 추출물로부터 새로운 2종류의 항균활성 펩타이드의 정제)

  • Go, Hye-Jin;Bae, Yun Jung;Park, Nam Gyu
    • Journal of Life Science
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    • v.24 no.8
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    • pp.860-864
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    • 2014
  • PAP-1, a novel antimicrobial peptide isolated from pyloric caeca extract of the starfish Asterina pectinifera was purified and characterized. First, the acidified pyloric caeca extract was put through Sep-Pak C18 solid phase extraction cartridge using a stepwise gradient. Among the eluents, RM 60 (retained materials at 60% methanol) showed good antimicrobial activity against Bacillus subtilis and Escherichia coli D31 and was purified in C18 reversed-phase and ion-exchange high-performance liquid chromatography columns. The purification steps yielded two novel peptides showing strong antimicrobial activities. These peptides were named pyloric caeca A. pectinifera peptide 1 and 2 (PAP-1 and PAP-2). For the characterization of the purified peptides, the molecular weights and amino acid sequences were determined by MALDI-TOF MS and Edman degradation. The molecular weights of PAP-1 and PAP-2 were about 2951.54 Da and 2980.15 Da respectively. The amino acid sequences of PAP-1 and PAP-2 were partially determined: AIQNAGES and AIQNAAES, respectively. PAP-2 is an isoform of PAP-1, differing merely by a single residue at position 6 (glycine or alanine). The comparison of the N-terminal amino acid sequences and molecular weights of the peptides with those of other known antimicrobial peptides revealed that PAP-1 and PAP-2 have no homology with any known peptides. These findings suggest that PAP-1 and PAP-2 play a significant role in the innate defense system of starfish pyloric caeca.

Utilization of Pigments and Tunic Components of Ascidian as an Improved Feed Aids for Aquaculture 2. Chemical Properties of Sulfated Polysaccharides in Ascidian (Halocynthia roretzi) Tunic (우렁쉥이 껍질성분 및 색소를 이용한 양식소재 개발 2. 우렁쉥이 껍질 중 부분정제한 황산다당의 화학적 특성)

  • HONG Byeong-Il;JUNG Byung-Chun;JUNG Woo-Jin;RUCK Ji-Hee;CHOI Byeong-Dae;LEE Kang-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.34 no.6
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    • pp.632-637
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    • 2001
  • Components of polysaccharides isolated from ascidian tunic were measuerd by gel filtration, electrophoresis and chemical analyses. The sulfated polysaccharides consisted in sulfate, protein, uronic acid and amino sugars. Hexosamines were composed of arabinose, xylose, glucose, galactose, glucuronic acid, N-acetylgalactosamine and N-acetylglucosamine by gas chromatography analysis. The galactose was predominant hexose after autoclave and nutrase digestion followed by DEAE-cellulose ion exchange chromatography and gel-permeation chromatography on Sephadex G-100 and G-25. FT-IR spectra of isolated polysaccharides from ascidian tunic and standard chondroitin sulfates have similar functional groups of the type of vibration and frequency. Molecular weight of isolated polysaccharides by autoclave represented more than 40 kDa by polyacrylamide gel electrophoresis. But the neutrase treatment group divided into three band. The highest molecular band group was shown more than 100 kDa, and the two low molecular band group were shown about 22 kDa and 5 kDa, respectively, compare to standard materials.

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Serodiagnosis of human paragonimiasis by ELISA-inhibition test using monoclonal antibodies (단클론항체를 이용한 폐흡충증의 면역진단)

  • Yong, Tae-Sun;Seo, Jang-Hun;Yeo, In-Seok
    • Parasites, Hosts and Diseases
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    • v.31 no.2
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    • pp.141-148
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    • 1993
  • ELISA-inhibition test using Paragonimus westermani specific monoclonal antibody (Mab) was investigated to improve the diagnostic specificity of paragonimiasis. By cell fusion, one hybridoma clone secreting un-n westemanl specific Mab was selected (Pwa-14), which reacted on bands of 28 kDa, 42.5 kDa, 89 kDa and 120.5 kDa. IFA showed Pwa-14 was located at the vitelline follicles. By micro-ELISA, 100% of 22 paragonimiasis cases were found positive, but 5 of 40 clonorchlasls cases (12.5%),3 of 26 cystlcercosis cases (7.7%) showed false positive. None of 10 sparganosis patients or 28 normal controls reacted positively. On the other hand, by ELISA-Inhibition test using a R westermcni specific Mab, 100% of patagonimlasls cases were found positive, and there were no positive in cysticercosis, sparganosis cases or normal controls, except 2 (5.0%) false-positive sera of 40 clonorchiasis cases. The ELISA-Inhlbltlon test using a Mab showed higher specificity in comparison with macro-ELISA for serodlgnosis of human paragonimlasis.

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Growth and Characterization of I $n_{x}$G $a_{1-x}$N Epitaxial Layer for Blue Light Emitter (청색발광소자를 위한 I $n_{x}$G $a_{1-x}$N 결정성장 및 특성평가)

  • 이숙헌;이제승;허정수;이병규;이승하;함성호;이용현;이정희
    • Journal of the Korean Institute of Telematics and Electronics D
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    • v.35D no.8
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    • pp.15-23
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    • 1998
  • Single crystalline I $n_{x}$G $a_{1-x}$ N thin film was grwon by MOCVD on (001) sapphire substrate for the blue light emitting devices. A good quality of I $n_{0.13}$G $a_{0.87}$N/GaN heterostructure grwon above 700.deg. C was confiremed by various characterization techniques of AFM, RHEED and DC-XRD. Through PL measurement at room temperautre for the Si-Zn co-doped I $n_{x}$G $a_{a-x}$N/GaN structure grwon at 800.deg. C to obtain blue wavelength emission, 460-470 nm and 425 nm emission peak were observed, which are believed to be from donor-to-acceptor pair transition and band edge emission of In/x/G $a_{1-x}$ N, respectively. The result of PL measurement of the undoped MQW I $n_{x}$G $a_{1-x}$ N layer at low temperature confirmed that the strong MQW peak was resulted by exciton from the GAN barrier and carrier of DA pair confined into the well layer.ll layer.yer.r.

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Discrimination model of cultivation area of Corni Fructus using a GC-MS-Based metabolomics approach (GC-MS 기반 대사체학 기법을 이용한 산수유의 산지판별모델)

  • Leem, Jae-Yoon
    • Analytical Science and Technology
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    • v.29 no.1
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    • pp.1-9
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    • 2016
  • It is believed that traditional Korean medicines can be managed more scientifically through the development of logical criteria to verify their region of cultivation, and that this could contribute to the advancement of the traditional herbal medicine industry. This study attempted to determine such criteria for Sansuyu. The volatile compounds were obtained from 20 samples of domestic Corni fructus (Sansuyu) and 45 samples of Chinese Sansuyu by steam distillation. The metabolites were identified in the NIST Mass Spectral Library via the obtained gas chromatography/mass spectrometer (GC/MS) data of 53 training samples. Data binning at 0.2 min intervals was performed to normalize the number of variables used in the statistical analysis. Multivariate statistical analyses, such as principle component analysis (PCA), partial least squares-discriminant analysis (PLS-DA), and orthogonal partial least squares-discriminant analysis (OPLS-DA) were performed using the SIMCA-P software package. Significant variables with a variable importance in the projection (VIP) score higher than 1.0 were obtained from OPLS-DA, and variables that resulted in a p-value of less than 0.05 through one-way ANOVA were selected to verify the marker compounds. Finally, among the 11 variables extracted, 1-ethylbutyl-hydroperoxide (9.089 min), nonadecane (20.170 min), butylated hydroxytoluene (25.319 min), 5β,7βH,10α-eudesm-11-en-1α-ol (25.921 min), 7,9-bis(2-methyl-2-propanyl)-1-oxaspiro[4.5]deca-6,9-diene-2,8-dione (34.257 min), and 2-decyldodecyl-benzene (54.717 min) were selected as markers to indicate the origin of Sansuyu. The statistical model developed was suitable for the determination of the geographical origin of Sansuyu. The cultivation areas of four Korean and eight Chinese Sansuyu samples were predicted via the established OPLS-DA model, and it was confirmed that 11 of the 12 samples were accurately classified.

On the Statistical Properties of the Parameters B and q in Creep Crack Growth Law, da/dt=B(C*)q, in the Case of Mod. 9Cr-1Mo Steel (Mod. 9Cr-1Mo강의 크리프 균열 성장 법칙의 파라메터 B와 q의 통계적 성질에 관한 연구)

  • Kim, Seon-Jin;Park, Jae-Young;Kim, Woo-Gon
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.35 no.3
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    • pp.251-257
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    • 2011
  • This paper deals with the statistical properties of parameters B and q in the creep crack growth rate (CCGR) law, da/dt=B$(C^*)^q$, in Mod. 9Cr-1Mo (ASME Gr.91) steel which is considered a candidate materials for fabricating next generation nuclear reactors. The CCGR data were obtained by creep crack growth (CCG) tests performed on 1/2-inch compact tension (CT) specimens under an applied load of 5000N at a temperature of $600^{\circ}C$. The CCG behavior was analyzed statistically using the empirical equation between CCGR, da/dt and the creep fracture mechanics parameter, $C^*$. The B and q values were determined for each specimen by the least-squares fitting method. The probability distribution functions for B and q were investigated using normal, log-normal, and Weibull distributions. As far as this study is considered, it can be appeared that B and q followed the log-normal and Weibull distributions. Moreover, a strong positive linear correlation was found between B and q.