• Title/Summary/Keyword: Ni-substrate

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Enzymatic characterization and Expression of 1-aminocycloprophane-1-carboxlyate deaminase from the rhizobacterium Pseudomonas flourescens

  • Lee, Gun-Woong;Ju, Jae-Eun;Kim, Hae-Min;Lee, Si-Nae;Chae, Jong-Chan;Lee, Yong-Hoon;Oh, Byung-Taek;Soh, Byoung-Yul
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2010.05a
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    • pp.17-17
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    • 2010
  • Ethylene, known as a stress hormone regulate wide developmental processes including germination, root hair initiation, root and shoot primordial formation and elongation, leaf and flower senescence and abscission, fruit ripening. The acceleration of ethylene biosynthesis in plant associated with environmental and biological stresses. 1-Aminocycloprophane-1-carboxlyate deaminase(ACCD) is an enzyme that cleaves ACC into and ammonia, a precursor of the plant hormone ethylene. Plant growth-promoting rhizobacteria (PGPR) having ACCD can decrease endogenous ACC level of tissue, resulting in reduced production of ethylene in plants. ACC deaminse was a key enzyme for protect stressed plants from injurious effects of ethylene. ACCD gene was encoded from Pseudomonas flourescens, PGPR and was cloned in Escherichia coli. We expressed the recombinant ACCD(rACCD) containing 357 amino acids with molecular weight 39 kDa that revealed by SDS-PAGE and western blot. The rACCD was purified by Ni-NTA purification system. The active form of rACCD having enzyme activity converted ACC to a-ketobutyrate. The optimal pH for ACC deaminase activity was pH 8.5, but no activity below pH 7.0 and a less severe tapering activity at base condition resulting in loss of activity at over pH 11. The optimal temperature of the enzyme was $30^{\circ}$ and a slightly less severe tapering activity at 15 - 30$^{\circ}$, but no activity over $35^{\circ}$. P. flourescens ACC deaminase has a highly conserved residue that plays in allowing substrate accessibility to the active sites. The enzymatic properties of this rACCD will provide an important reference for analysis of newly isolated ACCD and identification of newly isolated PGPR containing ACCD.

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Molecular Cloning and Expression of a Novel Protease-resistant GH-36 $\alpha$-Galactosidase from Rhizopus sp. F78 ACCC 30795

  • Yanan, Cao;Wang, Yaru;Luo, Huiying;Shi, Pengjun;Meng, Kun;Zhou, Zhigang;Zhang, Zhifang;Yao, Bin
    • Journal of Microbiology and Biotechnology
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    • v.19 no.11
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    • pp.1295-1300
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    • 2009
  • A 2,172-bp full-length gene (aga-F78), encoding a protease-resistant $\alpha$-galactosidase, was cloned from Rhizopus sp. F78 and expressed in Escherichia coli. The deduced amino acid sequence shared highest identity (45.0%) with an $\alpha$-galactosidase of glycoside hydrolase family 36 from Absidia corymbifera. After one-step purification with a Ni-NTA chelating column, the recombinant Aga-F78 migrated as a single band of ~82 and ~210 kDa on SDS-PAGE and nondenaturing gradient PAGE, respectively, indicating that the native structure of the recombinant Aga-F78 was a trimer. Exhibiting the similar properties as the authentic protein, purified recombinant Aga-F78 was optimally active at $50^{\circ}C$ and pH 4.8, highly pH stable over the pH range 5.0-10.0, more resistant to some cations and proteases, and had wide substrate specificity (pNPG, melidiose, raffinose, and stachyose). The recombinant enzyme also showed good hydrolytic ability to soybean meal, releasing galactose of $415.58\;{\mu}g/g$ soybean meal. When combined with trypsin, the enzyme retained over 90% degradability to soybean meal. These favorable properties make Aga-F78 a potential candidate for applications in the food and feed industries.

InGaN/GaN Blue LED device 제조시 ALD (Atomic Layer Deposition) 방법으로 증착된 Al2O3 Film의 Passivation 효과

  • Lee, Seong-Gil;Bang, Jin-Bae;Yang, Chung-Mo;Kim, Dong-Seok;Lee, Jeong-Hui
    • Proceedings of the Korean Vacuum Society Conference
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    • 2010.08a
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    • pp.211-212
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    • 2010
  • GaN 기반의 상부발광형 LED는 동작되는 동안 생기는 전기적 단락, 그리고 칩 위의 p-형 전극과 n-형 전극 사이에 생기는 누설전류 및 신뢰성 확보를 위하여 칩 표면에 passivation 층을 형성하게 된다. SiO2, Si3N4와 같은 passivation layers는 일반적으로 PECVD (Plasma Enhanced Chemical Vapor Deposition)공정을 이용한다, 하지만 이는 공정 특성상 plasma로 인한 damage가 유발되기 때문에 표면 누설 전류가 증가 한다. 이로 인해 forward voltage와 reverse leakage current의 특성이 저하된다. 본 실험에서는 원자층 단위의 박막 증착으로 인해 PECVD보다 단차 피복성이 매우 우수한 PEALD(Plasma Enhanced Atomic Layer Deposition)공정을 이용하여 Al2O3 passivation layer를 증착한 후, 표면 누설전류와 빛의 출력 특성에 대해서 조사해 보았다. PSS (patterned sapphire substrate) 위에 성장된 LED 에피구조를 사용하였고, TCP(Trancformer Copled Plasma)장비를 사용하여 에칭 공정을 진행하였다. 이때 투명전극을 증착하기 위해 e-beam evaporator를 사용하여 Ni/Au를 각각 $50\;{\AA}$씩 증착한 후 오믹 특성을 향상시키기 위하여 $500^{\circ}C$에서 열처리를 해주었다. 그리고 Ti/Au($300/4000{\AA}$) 메탈을 사용하여 p-전극과 n-전극을 형성하였다. Passivation을 하지 않은 경우에는 reverse leakage current가 -5V 에서 $-1.9{\times}10-8$ A 로 측정되었고, SiO2와 Si3N4을 passivation으로 이용한 경우에는 각각 $8.7{\times}10-9$$-2.2{\times}10-9$로 측정되었다. Fig. 1 에서 보면 알 수 있듯이 5 nm의 Al2O3 film을 passivation layer로 이용할 경우 passivation을 하지 않은 경우를 제외한 다른 passivation 경우보다 reverse leakage current가 약 2 order ($-3.46{\times}10-11$ A) 정도 낮게 측정되었다. 그 이유는 CVD 공정보다 짧은 ALD의 공정시간과 더 낮은 RF Power로 인해 plasma damage를 덜 입게 되어 나타난 것으로 생각된다. Fig. 2 에서는 Al2O3로 passivation을 한 소자의 forward voltage가 SiO2와 Si3N4로 passivation을 한 소자보다 각각 0.07 V와 0.25 V씩 낮아지는 것을 확인할 수 있었다. 또한 Fig. 3 에서는 Al2O3로 passivation을 한 소자의 output power가 SiO2와 Si3N4로 passivation을 한 소자보다 각각 2.7%와 24.6%씩 증가한 것을 볼 수 있다. Output power가 증가된 원인으로는 향상된 forward voltage 및 reverse에서의 leakage 특성과 공기보다 높은 Al2O3의 굴절률이 광출력 효율을 증가시켰기 때문인 것으로 판단된다.

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Fabrication and Characterization of an Antistiction Layer by PECVD (plasma enhanced chemical vapor deposition) for Metal Stamps (PECVD를 이용한 금속 스탬프용 점착방지막 형성과 특성 평가)

  • Cha, Nam-Goo;Park, Chang-Hwa;Cho, Min-Soo;Kim, Kyu-Chae;Park, Jin-Goo;Jeong, Jun-Ho;Lee, Eung-Sug
    • Korean Journal of Materials Research
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    • v.16 no.4
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    • pp.225-230
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    • 2006
  • Nanoimprint lithography (NIL) is a novel method of fabricating nanometer scale patterns. It is a simple process with low cost, high throughput and resolution. NIL creates patterns by mechanical deformation of an imprint resist and physical contact process. The imprint resist is typically a monomer or polymer formulation that is cured by heat or UV light during the imprinting process. Stiction between the resist and the stamp is resulted from this physical contact process. Stiction issue is more important in the stamps including narrow pattern size and wide area. Therefore, the antistiction layer coating is very effective to prevent this problem and ensure successful NIL. In this paper, an antistiction layer was deposited and characterized by PECVD (plasma enhanced chemical vapor deposition) method for metal stamps. Deposition rates of an antistiction layer on Si and Ni substrates were in proportion to deposited time and 3.4 nm/min and 2.5 nm/min, respectively. A 50 nm thick antistiction layer showed 90% relative transmittance at 365 nm wavelength. Contact angle result showed good hydrophobicity over 105 degree. $CF_2$ and $CF_3$ peaks were founded in ATR-FTIR analysis. The thicknesses and the contact angle of a 50 nm thick antistiction film were slightly changed during chemical resistance test using acetone and sulfuric acid. To evaluate the deposited antistiction layer, a 50 nm thick film was coated on a stainless steel stamp made by wet etching process. A PMMA substrate was successfully imprinting without pattern degradations by the stainless steel stamp with an antistiction layer. The test result shows that antistiction layer coating is very effective for NIL.

Fabrication of Environmental-friendly Materials Using Atomic Layer Deposition (원자층 증착을 이용한 친환경 소재의 제조)

  • Kim, Young Dok
    • Applied Chemistry for Engineering
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    • v.23 no.1
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    • pp.1-7
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    • 2012
  • In this article, I will introduce recent developments of environmental-friendly materials fabricated using atomic layer deposition (ALD). Advantages of ALD include fine control of the thin film thickness and formation of a homogeneous thin fim on complex-structured three-dimensional substrates. Such advantages of ALD can be exploited for fabricating environmental-friendly materials. Porous membranes such as anodic aluminum oxide (AAO) can be used as a substrate for $TiO_2$ coating with a thickness of about 10 nm, and the $TiO_2$-coated AAO can be used as filter of volatile organic compound such as toluene. The unique structural property of AAO in combination with a high adsorption capacity of amorphous $TiO_2$ can be exploited in this case. $TiO_2$ can be also deposited on nanodiamonds and Ni powder, which can be used as photocatalyst for degradation of toluene, and $CO_2$ reforming of methane catalyst, respectively. One can produce structures, in which the substrates are only partially covered by $TiO_2$ domains, and these structures turns out to be catalytically more active than bare substrates, or complete core-shell structures. We show that the ALD can be widely used not only in the semiconductor industry, but also environmental science.

Purification and Characterization of Gibberellin $3Beta$-Hydroxylase from Immature Seeds of Phaseolus vulgaris (강낭콩미숙종자로부터 Gibberellin $3Beta$-Hydroxylase 정제 및 성질)

  • 곽상수
    • Proceedings of the Botanical Society of Korea Conference
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    • 1987.07a
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    • pp.133-148
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    • 1987
  • Gibberellin(GA) 3-$\beta$ hydroxylation is very important for the shoot elogation in the higher plants, since only 3$\beta$-hydryoxylated GAs promote shoot elogation in several plants. Fluctuation of 3$\beta$-hydryoxylase activity was examined during seed maturation using two cultivars of , P. vulgaris, Kentucky Wonder (normal) and Masterpiece (dwarf). Very immature seeds of both cultivars contain high level of 3$\beta$-hydroxylase activity (per mg protein). Both cultivars showed maximum of enzyme activity (per seed) in the middle of their maturation process. Gibberellin 3$\beta$-hydroxylase catalyzing the hydroxylation of GA20 to GA1 was purified 313-fold from very early immature seeds of P. vulgaris. Crude soluble enzyme extracts were purified by 15% methanol precipitation, hydrophobic interaction chromatogrphy, DEAE ion exchange column chromatography and gel filtration HPLC. The 3$\beta$-hydroxylase activity was unstable and lost much of its activity duting the purification. The molecular weight of purified enzyme was extimated to be 42, 000 by gel filtration HPLC and SDS-PAGE. The enzyme exhibited maximum activity at pH 7.7. The Km values for [2.3-3H] GA20 and [2.3-3H]GA9 were 0.29 $\mu$M and 0.33 $\mu$M, respectively. The enzyme requires 2-oxoglutarate as a cosubstrate; the Km value for 2-oxoglutarate was 250 $\mu$M using 3H GA20 as a substrate. Fe2+ and ascorbate significantly activated the enzyme at all purification steps, while catalase and BSA activated the purified enzyme only. The enzyme was inhibited by divalent cations Mn2+, Co2+, Ni2+, Cu2+, Zn2+, Cd2+ and Hg2+. Effects of several GAs and GA anaogues on the putrified 3$\beta$-hydroxylase were examined using [3H]GA9 and GA20 as a substrates. Among them, GA5, GA9, GA15, GA20 and GA44 inhibited the enzyme activity. [13C, 3H] GA20 was converted by the partially purified enzyme preparation to [13C, 3H]GA1, GA5 and GA6, which were identified by GC-MS, GA9 was converted only GA4, GA15 and GA44 were converted to GA37 and GA38, respectively. GA5 was epoxidized to GA6 by the preparation. This suggests that 3$\beta$-hydroxylation of GA20 and epoxidation of GA5 are catalyzed by the same enzyme in P, vulgaris.

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Cloning and Expression of Thermostable $\beta$-Glycosidase Gene from Thermus filiformis Wai33 A1 in Escherichia coli and Enzyme Characterization

  • Kang, Sang-Kee;Cho, Kwang-Keun;Ahn, Jong-Kun;Kang, Seung-Ha;Han, Kyung-Ho;Lee, Hong-Gu;Choi, Yun-Jaie
    • Journal of Microbiology and Biotechnology
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    • v.14 no.3
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    • pp.584-592
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    • 2004
  • A thermostable $\beta$-glycosidase gene, tfi $\beta$-gly, was cloned from the genomic library of Thermus filiformis Wai33 A1. ifi $\beta$-gly consists of 1,296 bp nucleotide sequence and encodes a polypeptide of 431 amino acids. It shares a strong amino acid sequence similarity with the $\beta$-glycosidases from other Thermus spp. belonging to the glycosyl hydrolase family 1. In the present study, the enzyme was overexpressed in Escherichia coli BL21 (DE3) using the pET21b(+) vector system. The recombinant enzyme was purified to homogeneity by heat treatment and a $Ni^{2+}$-affinity chromatography. Polyacrylamide gel electrophoresis (PAGE) showed that the recombinant Tfi $\beta$-glycosidase was a monomeric form with molecular mass of 49 kDa. The temperature and pH range for optimal activity of the purified enzyme were 80- $90^{\circ}C$ and 5.0-6.0, respectively. Ninety-three percent of the enzyme activity was remained at $70^{\circ}C$ after 12 h, and its half-life at $80^{\circ}C$ was 6 h, indicating that Tfi $\beta$-glycosidase is highly thermostable. Based on its K_m$, or $K_{cat}K_m$, ratio, Tfi $\beta$-glycosidase appeared to have higher affinity for $\beta$-D-glucoside than for $\beta$-D-galactoside, however, $K_{cat} for \beta$-D-galactoside was much higher than that for $\beta$-D-glucoside. The activity for lactose hydrolysis was proportionally increased at $70^{\circ}C$ and pH 7.0 without substrate inhibition until reaching 250 mM lactose concentration. The specific activity of Tfi TEX>$\beta$-glycosidase on 138 mM lactose at $70{^\circ}C$ and pH 7.0 was 134.9 U/mg. Consequently, this newly cloned enzyme appears to have a valuable advantage of conducting biotechnological processes at elevated temperature during milk pasteurization in the production of low-lactose milk.

Evaluations of Si based ternary anode materials by using RF/DC magnetron sputtering for lithium ion batteries

  • Hwang, Chang-Muk;Park, Jong-Wan
    • Proceedings of the Korean Vacuum Society Conference
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    • 2010.08a
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    • pp.302-303
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    • 2010
  • Generally, the high energy lithium ion batteries depend intimately on the high capacity of electrode materials. For anode materials, the capacity of commercial graphite is unlike to increase much further due to its lower theoretical capacity of 372 mAhg-1. To improve upon graphite-based negative electrode materials for Li-ion rechargeable batteries, alternative anode materials with higher capacity are needed. Therefore, some metal anodes with high theoretic capacity, such as Si, Sn, Ge, Al, and Sb have been studied extensively. This work focuses on ternary Si-M1-M2 composite system, where M1 is Ge that alloys with Li, which has good cyclability and high specific capacity and M2 is Mo that does not alloy with Li. The Si shows the highest gravimetric capacity (up to 4000mAhg-1 for Li21Si5). Although Si is the most promising of the next generation anodes, it undergoes a large volume change during lithium insertion and extraction. It results in pulverization of the Si and loss of electrical contact between the Si and the current collector during the lithiation and delithiation. Thus, its capacity fades rapidly during cycling. Si thin film is more resistant to fracture than bulk Si because the film is firmly attached to the substrate. Thus, Si film could achieve good cycleability as well as high capacity. To improve the cycle performance of Si, Suzuki et al. prepared two components active (Si)-active(Sn, like Ge) elements film by vacuum deposition, where Sn particles dispersed homogeneously in the Si matrix. This film showed excellent rate capability than pure Si thin film. In this work, second element, Ge shows also high capacity (about 2500mAhg-1 for Li21Ge5) and has good cyclability although it undergoes a large volume change likewise Si. But only Ge does not use the anode due to its costs. Therefore, the electrode should be consisted of moderately Ge contents. Third element, Mo is an element that does not alloys with Li such as Co, Cr, Fe, Mn, Ni, V, Zr. In our previous research work, we have fabricated Si-Mo (active-inactive elements) composite negative electrodes by using RF/DC magnetron sputtering method. The electrodes showed excellent cycle characteristics. The Mo-silicide (inert matrix) dispersed homogeneously in the Si matrix and prevents the active material from aggregating. However, the thicker film than $3\;{\mu}m$ with high Mo contents showed poor cycling performance, which was attributed to the internal stress related to thickness. In order to deal with the large volume expansion of Si anode, great efforts were paid on material design. One of the effective ways is to find suitably three-elements (Si-Ge-Mo) contents. In this study, the Si based composites of 45~65 Si at.% and 23~43 Ge at.%, and 12~32 Mo at.% are evaluated the electrochemical characteristics and cycle performances as an anode. Results from six different compositions of Si-Ge-Mo are presented compared to only the Si and Ge negative electrodes.

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Effects of Non-ionic or Zwitterionic Surfactant on in vitro Digestibility of Rice Straw and Growth of Rumen Mixed Microorganisms. (비이온성 및 양쪽 이온성 계면활성제 첨가가 반추위 혼합 미생물의 성장과 볏짚의 in vitro 소화에 미치는 영향)

  • Lee, Shin-Ja;Kim, Wan-Young;Moon, Yea-Hwang;Kim, Hyeon-Shup;Kim, Kyoung-Hoon;Ha, Jong-Kyu;Lee, Sung-Sill
    • Journal of Life Science
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    • v.18 no.4
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    • pp.515-521
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    • 2008
  • This experiment was conducted to investigate effects of non-ionic or zwitterionic (+/-) surfactants on digestibility of rice straw, and changes of growth of rumen mixed microbes, pH, and gas production during in vitro fermentation. Also, during in vitro ruminal fermentation, microbial attachment on rice straw was investigated using scanning electron microscopy (SEM). Tween 80 or SOLFA-850 for non-ionic surfactant (NIS), and 3-(Dodecyldimethylammonio) propanesulfanate (DDAP) for zwitterionic surfactant (ZIS) was supplemented by 0.05% and 0.1% in Dehority's artificial medium containing Holtein rumen fluid, respectively, and the substrate for fermentation was rice straw passed through 1 mm screen. The experiment was composed of 7 treatments (two levels of two NISs, two levels of a ZIS) including the control, and 6, 12, 24, 48 and 72 hr of fermentation time with 3 replications per treatment. Treatment of Tween 80 increased in vitro DM digestibilities during 48 hr and 72 h post fermentations compared to the other treatments, whereas treatment of DDAP as a ZIS resulted in decreased DM digestibility than that of the control from 24 hr post fermentation (P<0.05). Gas production in vitro was greater (P<0.05) with addition of NIS than the control or ZIS, and increased as fermentation time elapsed. Rumen mixed microbial growth was greatest with addition of Tween 80 as NIS, and lowest when DDAP as ZIS was supplemented to the fermentation tube (P<0.05). In SEM observation, rumen microbial population attached on rice straw particle was greater with addition of NIS, but was less with addition ZIS compared with the control. In conclusion we could not found any positive effects of ZIS surfactants on rumunal fermentation characteristics and rumen microbial growth rates.