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LC-MS/MS Analysis of Surface Layer Proteins as a Useful Method for the Identification of Lactobacilli from the Lactobacillus acidophilus Group

  • Podlesny, Marcin;Jarocki, Piotr;Komon, Elwira;Glibowska, Agnieszka;Targonski, Zdzislaw
    • Journal of Microbiology and Biotechnology
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    • v.21 no.4
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    • pp.421-429
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    • 2011
  • For precise identification of a Lactobacillus K1 isolate, LC-MS/MS analysis of the putative surface layer protein was performed. The results obtained from LTQ-FT-ICR mass spectrometry confirmed that the analyzed protein spot is the surface layer protein originating from Lb. helveticus species. Moreover, the identified protein has the highest similarity with the surface layer protein from Lb. helveticus R0052. To evaluate the proteomic study, multilocus sequence analysis of selected housekeeping gene sequences was performed. Combination of 16S rRNA sequencing with partial sequences for the genes encoding the RNA polymerase alpha subunit (rpoA), phenylalanyl-tRNA synthase alpha subunit (pheS), translational elongation factor Tu (tuf), and Hsp60 chaperonins (groEL) also allowed to classify the analyzed isolate as Lb. helveticus. Further classification at the strain level was achieved by sequencing of the slp gene. This gene showed 99.8% identity with the corresponding slp gene of Lb. helveticus R0052, which is in good agreement with data obtained by nano-HPLC coupled to an LTQ-FT-ICR mass spectrometer. Finally, LC-MS/MS analysis of surface layer proteins extracted from three other Lactobacillus strains proved that the proposed method is the appropriate molecular tool for the identification of S-layer-possessing lactobacilli at the species and even strain levels.

Genetically Engineered Biosynthesis of Macrolide Derivatives Including 4-Amino-4,6-Dideoxy-L-Glucose from Streptomyces venezuelae YJ003-OTBP3

  • Pageni, Binod Babu;Oh, Tae-Jin;Liou, Kwang-Kyoung;Yoon, Yeo-Joon;Sohng, Jae-Kyung
    • Journal of Microbiology and Biotechnology
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    • v.18 no.1
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    • pp.88-94
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    • 2008
  • Two sugar biosynthetic cassette plasm ids were used to direct the biosynthesis of a deoxyaminosugar. The pOTBP1 plasmid containing TDP-glucose synthase (desIII), TDP-glucose-4,6-dehydratase (desIV), and glycosyltransferase (desVII/desVIII) was constructed and transformed into S. venezuelae YJ003, a strain in which the entire gene cluster of desosamine biosynthesis is deleted. The expression plasmid pOTBP3 containing 4-aminotransferase (gerB) and 3,5-epimerase (orf9) was transformed again into S. venezuelae YJ003-OTBP1 to obtain S. venezuelae YJ003-OTBP3 for the production of 4-amino-4,6-dideoxy-L-glucose derivatives. The crude extracts obtained from S. venezuelae ATCC 15439, S. venezuelae YJ003, and S. venezuelae YJ003-OTBP3 were further analyzed by TLC, bioassay, HPLC, ESI/MS, LC/MS, and MS/MS. The results of our study clearly shows that S. venezuelae YJ003-OTBP3 constructs other new hybrid macrolide derivatives including 4-amino-4,6-dideoxy-L-glycosylated YC-17 (3, [M+ $Na^+$] m/z=464.5), methymycin (4, m/z=480.5), novamethymycin (6, m/z=496.5), and pikromycin (5, m/z=536.5) from a 12-membered ring aglycon (10-deoxymethynolide, 1) and a 14-membered ring aglycon (narbonolide, 2). These results suggest a successful engineering of a deoxysugar pathway to generate novel hybrid macrolide derivatives, including deoxyaminosugar.

Damage Classification by Time Density Function of Ultrasonic Pulse Signal occurred at Tire (타이어에서 발생하는 초음파펄스신호의 시간밀도함수에 의한 손상 분별)

  • Kang, Dae-Soo
    • The Journal of the Institute of Internet, Broadcasting and Communication
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    • v.15 no.6
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    • pp.291-296
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    • 2015
  • The tire damage classification method is researched on the periodicity detection of ramdomness ultrasonic signals to occur at the driving vehicle tire. Setting method of adaptive threshold is proposed in order to valid pulse detection by tire damage in ultrasonic noise on the road and used low pass filter for decrease signal ramdomness as preprocessing. Time interval of detected pulse is setted the density function depend on the vehicle's speed and the method of tire damage detection is proposed that measuring the first peak's time of time density function.The result of time density function in case of one damage material, the first peak's time is measured within the error limit of tire's rotation period, 169.8ms and 97.9ms and 81.8ms, about the speed of 50km/h and 80km/h and 100km/h. In case of more than one damage material, the sum of each peak's time is measured within the error limit of tire's rotation period about the speed.

Volatile Flavor Compounds of Saussurea lappa C.B. Clarke Root Oil by Hydro Distillation-GC and $GC/MS^+$

  • Chang, Kyung-Mi;Kim, Gun-Hee
    • Food Quality and Culture
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    • v.1 no.1
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    • pp.13-17
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    • 2007
  • The volatile flavor compounds of Saussurea lappa C.B. Clarke, a perennial, aromatic and medicinal herbaceous plant of the Asteraceae family, were isolated by the hydro distillation extraction method using a Clevenger-type apparatus, and analyzed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC/MS). The plant yielded a light yellow colored oil (0.02%, v/w). From S. lappa C.B. Clarke root oil, sixty-three volatile flavor compounds were tentatively identified, among which sesquiterpene was predominant (21.70%). The identified compounds of the root oil constituted 87.47% of the total peak area. From the constituents making up more than 5% of the volatile flavor components, a long-chain aldehyde, (7Z, 10Z, 13Z)-7, 10, 13-hexadecatrienal, was the most abundant volatile flavor compound (21.20%), followed by dehydrocostuslactone (10.30%) belonging to sesquiterpene lactone, valerenol (5.30%) and vulgarol B (5.06%).

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Modulated Sputtering System (MSS)을 이용한 박막 증착 및 분석

  • Kim, Dae-Cheol;Kim, Tae-Hwan;Kim, Yong-Hyeon;Han, Seung-Hui;Kim, Yeong-U
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.08a
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    • pp.192.1-192.1
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    • 2013
  • 본 연구는 기존의 Sputtering 방식에 Modulation 방식을 적용한 Modulated Sputtering System (MSS)에 관한 특성 관찰과 이를 이용한 박막 증착 및 분석에 관한 내용이다. MSS에 인가하는 전압은 pulse on 시간동안 타겟에 음의 전압이 인가되어 sputtering에 의한 박막이 증착되고, pulse off 시간동안에는 양의 전압을 인가하여 증착된 박막에 양이온을 입사시켜 에너지 전달에 의한 박막의 특성을 향상시키고 자한다. MSS에 인가되는 전압과 주파수, 그리고 펄스폭을 변화시키며 전압과 전류, 그리고 기판에 입사하는 이온에너지 특성을 관찰하였다. 또한 MSS를 이용하여 티타늄(Ti), 탄소(C), 알루미늄이 도핑된 산화아연(AZO) 박막을 증착하였다. 증착된 박막은 a-step, SEM, XRD, AFM, 4 point probe를 이용하여 박막의 두께, 결정성장면, 표면 거칠기, 비저항 등을 분석하였다. Ti 박막에서는 기판에 입사되는 양이온의 에너지가 증가함에 따라 결정 방위면이 (002)에서 (001)로 변화함을 확인하였고 탄소 박막과 AZO 박막의 경우에는 기판에 입사되는 양이온의 에너지 변화에 따라 박막의 전도도를 조절할 수 있음을 확인하였다.

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Effects of Expanding Feed Processing Technologies on Egg Characteristic and Nutrient Digestibility in Laying Hens (Expanding 사료 가공 기술이 산란계의 계란 품질 및 영양소 소화율에 미치는 효과)

  • Yoo J.S.;Chae S.J.;Park S.E.;Jang H.D.;Seok H.B.;Kim I.H.;Kim Y.C.
    • Korean Journal of Poultry Science
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    • v.33 no.2
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    • pp.127-131
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    • 2006
  • This study was conducted to investigate the effects of expanding feed processing technologies on the egg characteristics and nutrient digestibility in laying hens. A total of two hundred seventy laying hens were randomly allocated into three treatments with fifteen replications for six weeks. Dietary treatments include 1) MS(mash), 2) EC(expanded crumble) and 3) EP(expanded pellet). In the egg production, the EC and EP treatments were significantly higher than MS treatment(p<0.05). However, no significant difference was founded in egg weight(p>0.05). Egg shell breaking was increased in EC and EP treatments compared with MS treatment(p<0.05), however, egg shall thickness showed no significant difference(p>0.05). Egg production and egg shell breaking were significantly higher in EC and EP treatments than MS treatment(p<0.05), but egg weight and egg shell thickness were no significantly difference among treatment(p<0.05). Yolk color was increased in EC treatment compared with other treatments(p<0.05). Also, yolk index was significantly improved in EC and EP treatments compared with MS treatment(p<0.05). Haugh unit was increased in EC treatment compared with MS treatment(p<0.05). EC treatment had improved DM, Ca and P digestibility. In conclusion, Expanded crumble and pellet processing could improve the egg characteristic and nutrient digestibility in laying hens.

Thermal and Cholesteric Mesophase Properties of Hydroxypropyl Celluloses and (Butoxypropyl)celluloses (히드록시프로필 셀룰로오스들 그리고 (부톡시프로필)셀룰로오스들의 열 및 콜레스테릭 상의 특성)

  • Jeong, Seung-Yong;Ma, Yung-Dae
    • Polymer(Korea)
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    • v.33 no.3
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    • pp.254-262
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    • 2009
  • Nine kinds of hydroxypropyl celluloses (HPCs) with degree of substitution (DS) and molar substitution (MS) ranging from 2.10 to 2.71 and 2.3 to 6.7, respectively and seven kinds of fully butanoated HPCs (BPCs) based on the HPCs with $2.3\;{\le}\;MS\;{\le}\;6.7$ were synthesized, and the molecular characteristics of HPCs and the thermotropic liquid crystalline properties of the derivatives were investigated. MS was nearly equal to DS for small value of DS, but it became exceedly larger than DS for $DS{\gtrsim}1$, showing that in the later stages of reaction, propylene oxide preferentially adds to the side chains rather than the main chain. All the derivatives formed enantiotropic cholesteric phases with right-handed helical structures. The glass and clearing transition temperatures of both HPCs and BPCs were decreased with increasing MS. The optical pitches (${\lambda}_m'S$) of BPCs, as well as HPCs themselves, increased with increasing temperature. However, the ${\lambda}_m'S$ of both HPCs and BPCs at the same temperature increased with increasing MS. Moreover, the temperature dependence of ${\lambda}_m$ of HPCs was weaker than that of BPCs, suggesting that the helical twisting power of the cellulose chain highly depends on the length and chemical structure of the side chain introduced in cellulose chain.

Simultaneous Determination and Occurrences of Pharmaceuticals by Solid-Phase Extraction and Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) in Environmental Aqueous Samples

  • Koo, So-Hyun;Jo, Cheon-Ho;Shin, Sun-Kyoung;Myung, Seung-Woon
    • Bulletin of the Korean Chemical Society
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    • v.31 no.5
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    • pp.1192-1198
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    • 2010
  • Pharmaceuticals and personal care products (PPCPs) are emerging contaminants in the aquatic environment. Many pharmaceuticals are not completely removed during wastewater treatment, leading to their presence in wastewater treatment effluents, rivers, lakes, and ground water. Here, we developed analytical methods for monitoring ten pharmaceuticals from surface water by LC/ESI-MS/MS. For sample clean-up and extraction, MCX (mixed cation exchange) and HLB (hydrophilic-lipophilic balance) solid-phase extraction (SPE) cartridges were used. The limits of detection (LOD) in distilled water and the blank surface water were in the range of 0.006 - 0.65 and 1.66 - 45.05 pg/mL, respectively. The limits of quantitation (LOQ) for the distilled water and the blank surface water were in the range of 0.02 - 2.17 and 5.52 - 150.15 pg/mL, respectively. The absolute recoveries for fortified water samples were between 62.1% and 125.4%. Intra-day precision and accuracy for the blank surface water were 2.9% - 24.1% (R.S.D.) and -16.3% - 16.3% (bias), respectively. In surface wastewater near rivers, chlortetracycline and acetylsalicylic acid were detected frequently in the range of 0.017 - 5.404 and 0.029 - 0.269 ng/mL, respectively. Surface water near rivers had higher levels than surface water of domestic treatment plants.

A Mismatch-Insensitive 12b 60MS/s 0.18um CMOS Flash-SAR ADC (소자 부정합에 덜 민감한 12비트 60MS/s 0.18um CMOS Flash-SAR ADC)

  • Byun, Jae-Hyeok;Kim, Won-Kang;Park, Jun-Sang;Lee, Seung-Hoon
    • Journal of the Institute of Electronics and Information Engineers
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    • v.53 no.7
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    • pp.17-26
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    • 2016
  • This work proposes a 12b 60MS/s 0.18um CMOS Flash-SAR ADC for various systems such as wireless communications and portable video processing systems. The proposed Flash-SAR ADC alleviates the weakness of a conventional SAR ADC that the operation speed proportionally increases with a resolution by deciding upper 4bits first with a high-speed flash ADC before deciding lower 9bits with a low-power SAR ADC. The proposed ADC removes a sampling-time mismatch by using the C-R DAC in the SAR ADC as the combined sampling network instead of a T/H circuit which restricts a high speed operation. An interpolation technique implemented in the flash ADC halves the required number of pre-amplifiers, while a switched-bias power reduction scheme minimizes the power consumption of the flash ADC during the SAR operation. The TSPC based D-flip flop in the SAR logic for high-speed operation reduces the propagation delay by 55% and the required number of transistors by half compared to the conventional static D-flip flop. The prototype ADC in a 0.18um CMOS demonstrates a measured DNL and INL within 1.33LSB and 1.90LSB, with a maximum SNDR and SFDR of 58.27dB and 69.29dB at 60MS/s, respectively. The ADC occupies an active die area of $0.54mm^2$ and consumes 5.4mW at a 1.8V supply.

Physicochemical Quality Properties of Mudflat Solar Salt and Roasted Salt (갯벌천일염과 구운 소금의 이화학적 품질 특성)

  • Lee, Jung-Hee;Kim, Hag-Lyeol;Kim, In-Cheol
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.7
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    • pp.1048-1054
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    • 2014
  • The purpose of this study was carried out to investigate the cause of sulfur dioxide occurrence, general element composition, sulfur compounds, heavy metals, macro- & micro-minerals, and oxidation-reduction potential (ORP) following baking time course of RS (RS1, RS2, RS3, and RS4) and mudflat solar salts (MSS). Sulfur dioxide ($SO_2$) and sulfite ($SO{_3}^{2-}$) were not detected in MSS or RS. However, sulfate ($SO{_4}^{2-}$) content significantly decreased in RS (29,878.15~36,097.45 ppm) compared to that in MSS (35,601.65 ppm). ORP was 181.15 mV in MSS, and 58.55 mV in RS1. Moisture content was 9.34% in MSS and 0.00% in RS with increased NaCl (94.77~95.77%). Moisture and NaCl contents showed no significant difference in RS. Insoluble and sandy residues were higher in RS than in MSS, whereas Ca and K showed no significant difference. Mg and Cl contents were higher in RS than in MSS. Br level was higher in MSS (628.1 ppm) than in RS (512.72~586.62 ppm), but there was no significant difference in $NO_3$. Heavy metals (Pb, As, and Hg) were more abundant in RS than in MSS, but levels were still safe. These results suggest that MSS and RS may increase protection against from $SO_2$ and $SO{_3}^{2-}$.