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Isolation, Purification, and Characterization of a Thermostable Xylanase from a Novel Strain, Paenibacillus campinasensis G1-1

  • Zheng, Hongchen;liu, Yihan;Liu, Xiaoguang;Wang, Jianling;Han, Ying;Lu, Fuping
    • Journal of Microbiology and Biotechnology
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    • v.22 no.7
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    • pp.930-938
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    • 2012
  • High levels of xylanase activity (143.98 IU/ml) produced by the newly isolated Paenibacillus campinasensis G1-1 were detected when it was cultivated in a synthetic medium. A thermostable xylanase, designated XynG1-1, from P. campinasensis G1-1 was purified to homogeneity by Octyl-Sepharose hydrophobic-interaction chromatography, Sephadex G75 gel-filter chromatography, and Q-Sepharose ion-exchange chromatography, consecutively. By multistep purification, the specific activity of XynG1-1 was up to 1,865.5 IU/mg with a 9.1-fold purification. The molecular mass of purified XynG1-1 was about 41.3 kDa as estimated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Sequence analysis revealed that XynG1-1 containing 377 amino acids encoded by 1,134 bp genomic sequences of P. campinasensis G1-1 shared 96% homology with XylX from Paenibacillus campinasensis BL11 and 77%~78% homology with xylanases from Bacillus sp. YA-335 and Bacillus sp. 41M-1, respectively. The activity of XynG1-1 was stimulated by $Ca^{2+}$, $Ba^{2+}$, DTT, and ${\beta}$-mercaptoethanol, but was inhibited by $Ni^{2+}$, $Fe^{2+}$, $Fe^{3+}$, $Zn^{2+}$, SDS, and EDTA. The purified XynG1-1 displayed a greater affinity for birchwood xylan, with an optimal temperature of $60^{\circ}C$ and an optimal pH of 7.5. The fact that XynG1-1 is cellulose-free, thermostable (stability at high temperature of $70^{\circ}C{\sim}80^{\circ}C$), and active over a wide pH range (pH 5.0~9.0) suggests that the enzyme is potentially valuable for various industrial applications, especially for pulp bleaching pretreatment.

Omega-7 producing alkaliphilic diatom Fistulifera sp. (Bacillariophyceae) from Lake Okeechobee, Florida

  • Berthold, David Erwin;Rosa, Nina de la;Engene, Niclas;Jayachandran, Krish;Gantar, Miroslav;Laughinghouse, Haywood Dail IV;Shetty, Kateel G.
    • ALGAE
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    • v.35 no.1
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    • pp.91-106
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    • 2020
  • Incorporating renewable fuel into practice, especially from algae, is a promising approach in reducing fossil fuel dependency. Algae are an exceptional feedstock since they produce abundant biomass and oils in short timeframes. Algae also produce high-valued lipid products suitable for human nutrition and supplement. Achieving goals of producing algae fuels and high-valued lipids at competitive prices involves further improvement of technology, especially better control over cultivation. Manipulating microalgae cultivation conditions to prevent contamination is essential in addition to promoting optimal growth and lipid yields. Contamination of algal cultures is a major impediment to algae cultivation that can however be mitigated by choosing extremophile microalgae. This work describes the isolation of alkali-tolerant / alkaliphilic microalgae native to South Florida with ideal characteristics for cultivation. For that purpose, water samples from Lake Okeechobee were inoculated into Zarrouk's medium (pH 9-12) and incubated for 35 days. Selection resulted in isolation of three strains that were screened for biomass and lipid accumulation. Two alkali-tolerant algae Chloroidium sp. 154-1 and Chlorella sp. 154-2 were poor lipid accumulators. One of the isolates, the diatom Fistulifera sp. 154-3, was identified as a lipid accumulating, alkaliphilic organism capable of producing 0.233 g L-1 d-1 dry biomass and a lipid content of 20-30% dry weight. Lipid analysis indicated the most abundant fatty acid within Fistulifera sp. was palmitoleic acid (52%), or omega-7, followed by palmitic acid (17%), and then eicosapentanoic acid (15%). 18S rRNA phylogenetic analysis formed a well-supported clade with Fistulifera species.

Study of the Vibration and Shock Isolation for HEV Battery Pack (특수임무 차량 배터리 팩 진동/충격 저감 설계에 대한 연구)

  • Kim, Man-Dal;Jang, Duk-Jin;Lee, Sung-Jun;Hong, Seong-Wook
    • Journal of the Korean Society for Precision Engineering
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    • v.33 no.10
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    • pp.813-820
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    • 2016
  • Hybrid Electric Vehicles (HEVs) are developed to be operated with two kinds of power source (Diesel Engine and Electric Motor with Rechargeable High Voltage Battery Pack). HEVs for military vehicle require high reliability to provide stable powers under serious environment such as vibration and shock. To ensure normal operation of battery pack under serious environment such as vibration and shock, the high voltage battery pack needs to have appropriate dynamic characteristics. This paper presents a design procedure for high voltage battery pack with such characteristics. An isolator design is proposed to reduce vibration and shock. Associated random vibration and shock response of the high voltage battery pack are simulated under conditions suggested by MIL specifications. Its dynamic characteristics and vibration and shock responses are validated with experiments.

Novel Buck Mode Three-Level Direct AC Converter with a High Frequency Link

  • Li, Lei;Guan, Yue;Gong, Kunshan;Li, Guangqiang;Guo, Jian
    • Journal of Power Electronics
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    • v.18 no.2
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    • pp.407-417
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    • 2018
  • A novel family of Buck mode three-level direct ac converters with a high frequency link is proposed. These converters can transfer an unsteady high ac voltage with distortion into a regulated sinusoidal voltage with a low THD at the same frequency. The circuit configuration is constituted of a three-level converter, high frequency transformer, cycloconverter, as well as input and output filters. The topological family includes forward, push-pull, half-bridge, and full-bridge modes. In order to achieve a reliable three-level ac-ac conversion, and to overcome the surge voltage and surge current of the cycloconverter, a phase-shifted control strategy is introduced in this paper. A prototype is presented with experimental results to demonstrate that the proposed converters have five advantages including high frequency electrical isolation, lower voltage stress of the power switches, bi-directional power flow, low THD of the output voltage, and a higher input power factor.

High Power Factor Three-phase AC-DC Flyback Converter Module Using Zero Voltage Switching (영전압 스위칭을 이용한 고역률 3상 AC-DC Flyback 컨버터 모듈)

  • Lee, J.P.;Choi, J.Y.;Song, J.H.;Choy, I.;Yoon, T.Y.
    • Proceedings of the KIEE Conference
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    • 1999.07f
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    • pp.2701-2703
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    • 1999
  • A new mode of parallel operation of a modular 3-phase AC-DC Flyback converter for high power factor correction along with tight regulation is presented in this thesis. The converter offers input/output transformer isolation for safety, a unity input power factor for minimum reactive power, high efficiency and high power density for minimum weight and volume. Compared with previously developed 3-phase two-stage power converter, the advantage of the proposed converter does not require expensive high voltage and high current devices that are normally needed in popular boost type 3-phase converter. In this paper, a detailed small signal analysis of the modular 3-phase AC-DC flyback converter is provided for control purposes and also experimental results are included to confirm the validity of the analysis.

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A Study on the Development of High Stiffness Body for Suspension Performance (서스펜션 성능 확보를 위한 고강성 차페 개발 프로세스 연구)

  • Kim, Ki-Chang;Kim, Chan-Mook
    • Transactions of the Korean Society for Noise and Vibration Engineering
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    • v.15 no.7 s.100
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    • pp.799-805
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    • 2005
  • This paper describes the development process of high stiffness body for ride and handling performance. High stiffness and light weight vehicle is a major target in the refinement of Passenger cars to meet customers' contradictable requirements between ride and handling performance and fuel economy This paper describes the analysis approach process for high stiffness body through the data level of body stiffness. According to the frequency band. we can suggest the design guideline about lg cornering static stiffness, torsional and lateral stiffness, body attachment stiffness. The ride and handling characteristic of a vehicle Is significantly affected by vibration transferred to the body through the chassis mounting points from front and rear suspension. It is known that body attachment stiffness is an important factor of ride and handling performance improvement. And high stiffness helps to improve the flexibility of bushing rate tuning between handling and road noise. It makes possible to design the good handling performance vehicle and save vehicles to be used in tests by using mother car at initial design stage. These improvements can lead to shortening the time needed to develop better vehicles.

A Study on the Development of High Stiffness Body for Suspension Performance (서스펜션 성능 확보를 위한 고강성 차체 개발 프로세스 연구)

  • Kim, Ki-Chang;Kim, Chan-Mook
    • Proceedings of the Korean Society for Noise and Vibration Engineering Conference
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    • 2004.11a
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    • pp.358-361
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    • 2004
  • This paper describes the development process of high stiffness body for ride and handling performance. High stiffness and light weight vehicle is a major target in the refinement of passenger cars to meet customers' contradictable requirements between ride and handling performance and fuel economy. This paper describes the analysis approach process for high stiffness body through the data level of body stiffness. According to the frequency band, we can suggest the design guideline about Is cornering static stiffness, torsional and lateral stiffness, body attachment stiffness. The ride and handling characteristic of a vehicle is significantly affected by vibration transferred to the body through the chassis mounting points from front and rear suspension. It is known that body attachment stiffness is an important factor of ride and handling performance improvement. And high stiffness helps to improve the flexibility of bushing rate tuning between Handling and road noise. It makes it possible to design the good handling performance vehicle at initial design stage and save vehicles to be used in tests by using mother car at initial design stage. These improvements can lead to shortening the time needed to develop better vehicles.

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Studies on Protoplast Isolation of Pleurotus cornucopiae (노랑느타리버섯의 원형질체(原形質體) 분리(分離)에 관한 연구(硏究))

  • Lee, Yeon-Hee;Park, Yong-Hwan;Yoo, Young-Bok;Min, Kyung-Hee
    • The Korean Journal of Mycology
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    • v.14 no.2
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    • pp.141-148
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    • 1986
  • The optimal conditions for high yields of mycelial protoplasts from P. cornucopiae were established. The concentraion of enzyme system containing Novozym 234, ${\beta}-D-glucanase$ and ${\beta}-glucuronidase$ was $5mg\;ml^{-1}$ each. The osmotic stabilizer most effective for protoplast isolation was O.6 M sucrose. The optimal reaction time of mycelium with the lytic mixture was 90 min in a shaking condition at 120 strokes $min^{-1}$. When the myelium of P. cornucopiae was cultured for 4 days on mushroom complete medium at $28^{\circ}C$, the formation of protoplast was effective. When the pH of the digestion mixture with O.6 M sucrose as stabilizer varied between pH 4.0 and 7.0, the production of protoplasts was effective in phosphate buffer (pH 6.2) and Na-maleate buffer (pH 5.0). Generally, phosphate buffer was more effective for protoplast isolation than Na-maleate buffer, but 0.6 M sucrose osmotic stabilizer without adjusting pH was most effective. Using these conditions, protoplasts from P. cornucopiae were obtained at a ratio $1{\times}10^7\;ml^{-1}$.

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Studies on Protoplast Formation and Regeneration of Lyophyllum decastes (Lyophyllum decastes의 원형질체 분리와 재생에 관한 연구)

  • Bok, Jin-Woo;Kim, Jong-Pil;Jin, Mi-Rim;Choi, Eung-Chil;Kim, Byong-Kak
    • The Korean Journal of Mycology
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    • v.22 no.2
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    • pp.130-137
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    • 1994
  • This experiment was carried out to investigate proper conditions for protoplast isolation and regeneration from mycelia of Lyophyllum decastes. Novozym 234(10 mg/ml) with 0.6 M $MgSO_4$ in phosphate buffer(pH 4.0) was proper for protoplast isolation. The optimal reaction time of the mycelium with the lytic enzyme was four hours in shaking condition at 120 strokes per min. When the mycelium of L. decastes was cultured at $24^{\circ}C$ for 5 days, the formation of protoplasts was effective. The liquid medium was more effective for protoplast isolation than the solid medium. In the liquid medium, high yields of protoplasts were obtained from 0.6 M $MgSO_4$ osmotic stabilizer. Protoplasts of L. decastes were regenerated to normal hyphal growth and the regeneration frequency of the protoplasts in the complete agar medium containing Triton X-100(0.0025%) was $5.94{\sim}8.32%$. The regeneration medium stabilized with 0.6 M sucrose was the best for regeneration of the protoplasts. In contrast to protoplast formation, regeneration was inhibited by the inorganic salts used as osmotic stabilizer.

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Study on the Distribution of Vibrio parahaemolyticus along Cheju Coast (제주연안해역의 Vibrio parahaemolyticus 분포에 관한 연구)

  • Song, Min-Kyoung;Kim, Man-Chul;Heo, Moon-Soo
    • Korean Journal of Environmental Biology
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    • v.25 no.1
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    • pp.34-41
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    • 2007
  • A study on the distribution of V. parahaemolyticus among sea water, sea mud, and marine products in Hwabuk, Samyang, Daepo, Jungmun, Pyoson, Anduk, Aewol, and Gwakji on the coastal area of Jeju island was conducted from January to December in 2002. The 2,880 total specimens of 960 sea waters, 960 sea mud, 960 marine products were collected and studied for the rate of isolation of V. parahaemolyticus, and biochemical, serological and antibiotic sensitivity tests were performed. A total of 417 strains of V. parahaemolyticus were isolated and identified from 2,880 total specimens. In the test of biochemical properties, 100 of V. parahaemolyticus isolates in the presence of 0.85% NaCl were positive in the utilization of lysine, ornithine, indole, glucose, and mannitol, and negative in the utilization of ONPG, arginine, sodium citrate, urea, tryptophane, inositol, sorbitol, rhamnose, sucrose, and melibiose, $H_2S$ production and VP reaction, while positive or negative in gelatin liquefaction and utilization of amygdalin or arabinose. The isolation rates to the specimen were 161 strains (16.8%) from 960 of sea waters, 137 strains (14.3%) from 960 of sea mud, and 119 strains (12.4%) from 960 of marine products. The isolation rates of V. parahaemolyticus from 8 coastal areas were 14.4% (52/360) in Hwabuk area, 15.3% (55/360) in Samyang area, 13.6% (49/360) in Daepo area, 18.3% (66/360) Jungmun area, 13.1% (47/360) in Pyosun area, 16.4% (59/360) in Anduk area, 12.5% (45/360) in Aewol area and 12.2% (44/360) in Gwakji area, respectively. The distribution of 417 V. parahaemolyticus, isolates was high at Jungmun with 18.3% (66/360), and from sea water with 16.8% (161/960).