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Bioconversion of Ginsenosides from Red Ginseng Extract Using Candida allociferrii JNO301 Isolated from Meju

  • Lee, Sulhee;Lee, Yong-Hun;Park, Jung-Min;Bai, Dong-Hoon;Jang, Jae Kweon;Park, Young-Seo
    • Mycobiology
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    • v.42 no.4
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    • pp.368-375
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    • 2014
  • Red ginseng (Panax ginseng), a Korean traditional medicinal plant, contains a variety of ginsenosides as major functional components. It is necessary to remove sugar moieties from the major ginsenosides, which have a lower absorption rate into the intestine, to obtain the aglycone form. To screen for microorganisms showing bioconversion activity for ginsenosides from red ginseng, 50 yeast strains were isolated from Korean traditional meju (a starter culture made with soybean and wheat flour for the fermentation of soybean paste). Twenty strains in which a black zone formed around the colony on esculin-yeast malt agar plates were screened first, and among them 5 strains having high ${\beta}$-glucosidase activity on p-nitrophenyl-${\beta}$-D-glucopyranoside as a substrate were then selected. Strain JNO301 was finally chosen as a bioconverting strain in this study on the basis of its high bioconversion activity for red ginseng extract as determined by thin-layer chromatography (TLC) analysis. The selected bioconversion strain was identified as Candida allociferrii JNO301 based on the nucleotide sequence analysis of the 18S rRNA gene. The optimum temperature and pH for the cell growth were $20{\sim}30^{\circ}C$ and pH 5~8, respectively. TLC analysis confirmed that C. allociferrii JNO301 converted ginsenoside Rb1 into Rd and then into F2, Rb2 into compound O, Rc into compound Mc1, and Rf into Rh1. Quantitative analysis using high-performance liquid chromatography showed that bioconversion of red ginseng extract resulted in an increase of 2.73, 3.32, 33.87, 16, and 5.48 fold in the concentration of Rd, F2, compound O, compound Mc1, and Rh1, respectively.

Controlled Synthesis of Hexagonal Boron Nitride on Cu Foil Using Chemical Vapor Deposition

  • Han, Jaehyun;Lee, Jun-Young;Kwon, Heemin;Yeo, Jong-Souk
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.02a
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    • pp.630-630
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    • 2013
  • Recently, atomically smooth hexagonal boron nitride(h-BN) known as a white graphene has drawn great attention since the discovery of graphene. h-BN is a III-V compound and has a honeycomb structure very similar to graphene with smaller lattice mismatch. Because of strong covalent sp2bonds like graphene, h-BN provides a high thermal conductivity and mechanical strength as well as chemical stability of h-BN superior to graphene. While graphene has a high electrical conductivity, h-BN has a highly dielectric property as an insulator with optical band gap up to 6eV. Similar to the graphene, h-BN can be applied to a variety of field, such as gate dielectric layers/substrate, ultraviolet emitter, transparent membrane, and protective coatings. However, up until recently, obtaining and controlling good quality monolayer h-BN layers have been too difficult and challenging. In this work, we investigate the controlled synthesis of h-BN layers according to the growth condition, time, temperature, and gas partial pressure. h-BN is obtained by using chemical vapor deposition on Cu foil with ammonia borane (BH3NH3) as a source for h-BN. Scanning Transmission Electron Microscopy (STEM, JEOL-JEM-ARM200F) is used for imaging and structural analysis of h-BN layer. Sample's surface morphology is characterized by Field emission scanning electron microscopy (SEM, JEOL JSM-7100F). h-BN is analyzed by Raman spectroscopy (HORIBA, ARAMIS) and its topographic variations by Atomic force microscopy (AFM, Park Systems XE-100).

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Efficient Expression, Purification, and Characterization of a Novel FAD-Dependent Glucose Dehydrogenase from Aspergillus terreus in Pichia pastoris

  • Yang, Yufeng;Huang, Lei;Wang, Jufang;Wang, Xiaoning;Xu, Zhinan
    • Journal of Microbiology and Biotechnology
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    • v.24 no.11
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    • pp.1516-1524
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    • 2014
  • Flavin adenine dinucleotide-dependent glucose dehydrogenase (FAD-GDH) can utilize a variety of external electron acceptors and also has stricter substrate specificity than any other glucose oxidoreductases, which makes it the ideal diagnostic enzyme in the field of glucose biosensors. A gene coding for a hypothetical protein, similar to glucose oxidase and derived from Aspergillus terreus NIH2624, was overexpressed in Pichia pastoris GS115 under the control of an AOX1 promoter with a level of 260,000 U/l in the culture supernatant after fed-batch cultivation for 84 h. After a three-step purification protocol that included isopropanol precipitation, affinity chromatography, and a second isopropanol precipitation, recombinant FAD-GDH was purified with a recovery of 65%. This is the first time that isopropanol precipitation has been used to concentrate a fermentation supernatant and exchange buffers after affinity chromatography purification. The purified FAD-GDH exhibited a broad and diffuse band between 83 and 150 kDa. The recombinant FAD-GDH was stable across a wide pH range (3.5 to 9.0) with maximum activity at pH 7.5 and $55^{\circ}C$. In addition, it displayed very high thermal stability, with a half-life of 82 min at $60^{\circ}C$. These characteristics indicate that FAD-GDH will be useful in the field of glucose biosensors.

Anti-Corrosion Characteristics of Steel Structures with Polyaniline Anti-Corrosive Coatings (강 구조물에 대한 폴리아닐린 함유도료의 방청특성)

  • Song, Min-Kyung;Kong, Seung-Dae;Oh, Eun-Ha;Yoon, Hun-Cheol;Kim, Yoon-Shin;Im, Ho-Sub
    • Journal of Environmental Health Sciences
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    • v.36 no.3
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    • pp.236-246
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    • 2010
  • In preparative anti-corrosive coating experiments, polyaniline was obtained by reacting an oxidizing agent with the monomer aniline. Further, the primer coating was prepared using a variety of widely-used materials such as urethane resin. For the top coating, epoxy resin and acrylic urethane resin were used. Characteristics of the coatings were assessed according to KS and ASTM specifications, and the structure of the polyaniline was characterized using FT-IR and TGA. For analysis of anti-corrosive properties in salt-spray experiments, measurements of the oxidation state of iron and surface atomic analysis were conducted using XPS and SEM-EDX. Unlike general anti-corrosive coatings which exhibit anti-corrosive effects only as a primer coating, the anti-corrosive coatings using polyaniline as the anti-corrosive pigment showed a marked synergistic effect with the top coatings. In other words, the top coatings not only produce a fine view effect, but also increase, through interaction with the primer coatings, the resistance to diffusion of corrosive factors from the external environment. It was also found that, unlike the heavy metal oxide-forming layer of the passive barrier alone, the polyaniline anti-corrosive pigment oxidized iron at the interface with the iron substrate to form a passive barrier in the oxidic layer, and itself formed a potential barrier layer with anti-corrosive factors from the external environment. Although the passive layer was damaged, the damaged area did not become completely oxidized iron; on the contrary, it showed a tendency to reduction. This can be interpreted such that a passive layer is formed again on the damaged area, and that at the same time there is a tendency to self-healing.

Effects of Interleukin-1${\beta}$ and Tumor Necrosis $Factor-{\alpha}$ on the Release of Collagenase and Gelatinase from Osteoblasts

  • Eun, Jong-Gab;Baek, Dong-Heon;Kim, Se-Won
    • The Korean Journal of Physiology and Pharmacology
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    • v.6 no.5
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    • pp.269-274
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    • 2002
  • A large number of factors such as osteotropic hormones, cytokines, or growth factors are related to the bone remodeling which is characterized by the coupling of osteoclast-mediated bone resorption and osteoblast-mediated bone formation. Recent investigations have indicated that cytokines such as $interleukin-1{\beta}\;(IL-1{\beta})$ and tumor necrosis $factor-{\alpha}\;(TNF-{\alpha})$ play a potential role in the bone resorption associated with a variety of pathological conditions such as inflammatory osteolytic disease. Collagen is the most abundant protein of the extracellular matrix of bone, and the participation of collagenase in bone resorption has been widely investigated. In this study, effects of $IL-1{\beta}$ and $TNF-{\alpha}$ on the release of collagenase from osteoblastic cells were measured. The gelatinase activity was also measured by gel substrate analysis (zymography) after electrophoresis of conditioned media of osteoblastic cell culture. $IL-1{\beta}$ increased the collagenase activity in ROS17/2.8 and HOS cell culture. $TNF-{\alpha}$ also increased the collagenase activity of osteoblastic cells. When two kinds of cytokines were treated simultaneously in the culture of osteoblastic cells, synergistic increase of collagenase activity was seen in ROS17/2.8 cells. $IL-1{\beta}$ and $TNF-{\alpha}$ significantly increased the collagenase activity after 6 hour treatment in the osteoblastic cell culture, and there was no additional increase according to the culture period. Osteoblastic cells released the gelatinase and molecular weight of this enzyme was measured about 70 KDa as assessed by zymogram. $IL-1{\beta}$ and $TNF-{\alpha}$ showed increase of the gelatinase activity produced by ROS17/2.8 and HOS cells. Taken together, this study suggested that $IL-1{\beta}$ and $TNF-{\alpha}$ can modulate bone metabolism, at least in part, by increased release of collagenase and gelatinase from osteoblasts.

Electron Transport Carrier for the Free Radical Shethna Flavoprotein in Azotobacter vinelandii (Azotobacter vinelandii Shethna Flavoprotein 의 Free Radical 생성(生成)을 위(爲)한 전자전달물질(電子傳達物質)-)

  • Chun, Jae-Kun;Tollin, Gordon
    • Applied Biological Chemistry
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    • v.16 no.1
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    • pp.31-40
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    • 1973
  • Azotobacter vinelandii cell extracts and its variety of purified fractions with regard to their ability to form the redox state of the Shethna Flavoprotein (free radical form FPH.) were studied. A fluorescent flavoprotein (protein I) and a brown protein (protein II) were the most active proteins which were isolated in purified form. The free radical formation activity was substantially decreased during the purification and was completely lost upon storage in a week under nitrogen in a frozenstate. The presence of free flavin (FMN) with NADH enhanced the rate of free radical formation. The reaction of FMN and NADH was found to be catalysed by various cell fractions. A possible role of FMN as a substrate for free radical shethna flavoprotein was investigated. Slower reaction rate of $FMNH_2+Flavoprotein\;(FP){\to}FPH+FMN$ than $FMN+NADH{\to}FMNH_2$, accumulation of $FMNH_2$ ocurred which subsquently caused FPH.

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Hydrogen Absorption by Crystalline Semiconductors: Si(100), (110) and (111)

  • Jeong, Min-Bok;Jo, Sam-Geun
    • Proceedings of the Korean Vacuum Society Conference
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    • 2010.02a
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    • pp.383-383
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    • 2010
  • Gas-phase hydrogen atoms create a variety of chemical and physical phenomena on Si surfaces: adsorption, abstraction of pre-adsorbed H, Si etching, Si amorphization, and penetration into the bulk lattice. Thermal desorption/evolution analyses exhibited three distinct peaks, including one from the crystalline bulk. It was previously found that thermal-energy gaseous H(g) atoms penetrate into the Si(100) crystalline bulk within a narrow substrate temperature window(centered at ~460K) and remain trapped in the bulk lattice before evolving out at a temperature as high as ~900K. Developing and sustaining atomic-scale surface roughness, by H-induced silicon etching, is a prerequisite for H absorption and determines the $T_s$ windows. Issues on the H(g) absorption to be further clarified are: (1) the role of the detailed atomic surface structure, together with other experimental conditions, (2) the particular physical lattice sites occupied by, and (3) the chemical nature of, absorbed H(g) atoms. This work has investigated and compared the thermal H(g) atom absorptivity of Si(100), Si(111) and Si(110) samples in detail by using the temperature programmed desorption mass spectrometry (TPD-MS). Due to the differences in the atomic structures of, and in the facility of creating atom-scale etch pits on, Si(100), (100) and (110) surfaces, the H-absorption efficiency was found to be larger in the order of Si(100) > Si(111) > Si(110) with a relative ratio of 1 : 0.22 : 0.045. This intriguing result was interpreted in terms of the atomic-scale surface roughening and kinetic competition among H(g) adsorption, H(a)-by-H(g) abstraction, $SiH_3(a)$-by-H(g) etching, and H(g) penetraion into the crystalline silicon bulk.

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Optimized Decomposition of Ammonia Borane for Controlled Synthesis of Hexagonal Boron Nitride Using Chemical Vapor Deposition

  • Han, Jaehyu;Kwon, Heemin;Yeo, Jong-Souk
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.08a
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    • pp.285-285
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    • 2013
  • Recently, hexagonal boron nitride (h-BN), which is III-V compound of boron and nitride by strong covalent sp2 bonds has gained great interests as a 2 dimensional insulating material since it has honeycomb structure with like graphene with very small lattice mismatch (1.7%). Unlike graphene that is semi-metallic, h-BN has large band gap up to 6 eV while providing outstanding properties such as high thermal conductivity, mechanical strength, and good chemical stability. Because of these excellent properties, hBN can potentially be used for variety of applications such as dielectric layer, deep UV optoelectronic device, and protective transparent substrate. Low pressure and atmospheric pressure chemical vapor deposition (LPCVD and APCVD) methods have been investigated to synthesize h-BN by using ammonia borane as a precursor. Ammonia borane decomposes to polyiminoborane (BHNH), hydrogen, and borazine. The produced borazine gas is a key material that is a used for the synthesis of h-BN, therefore controlling the condition of decomposed products from ammonia borane is very important. In this paper, we optimize the decomposition of ammonia borane by investigating temperature, amount of precursor, and other parameters to fabricate high quality monolayer h-BN. Synthesized h-BN is characterized by Raman spectroscopy and its absorbance is measured with UV spectrophotometer. Topological variations of the samples are analyzed by atomic force microscopy. Scanning electron microscopy and Scanning transmission Electron microscopy are used for imaging and analysis of structures and surface morphologies.

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Analysis of lignocellulose degradation by Oak mushroom (Lentinula edodes) (원목재배용 표고(Lentinula edodes)의 목질섬유소 분해특성 비교)

  • Jeong, Sang-Wook;Jang, Eun-Gyeong;Jeong, Chan-Mun;Ko, Han-Gyu;Kwon, Hyuk-Woo;Ban, Seung-Eon
    • Journal of Mushroom
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    • v.16 no.4
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    • pp.272-278
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    • 2018
  • Lignin degrading enzymes from Lentinula edodes have broad substrate specificities, and therefore can degrade a variety of recalcitrant compounds. In this study, the lignolytic biodegradation was investigated in five different L. edodes fungi (Chunbaegko, Sanjo 303ho, Poongnyunko, Baekhwahyang, and Soohyangko). The fungi were evaluated for their ability to decolorize Remazol Brilliant Blue R (RBBR) in malt extract broth medium. Sanjo 303ho, Poongnyunko, Baekhwahyang, and Soohyangko rapidly decolorized RBBR within 7 days. The activities of manganese peroxidase (MnP) and laccase were determined in the absence and presence of lignin. Poongnyunko displayed the highest ligninolytic activity on day 7 of incubation (2,809 U/mg and 2,230 U/mg for MnP and laccase, respectively).

Preparation of Hypoallergenic Whey Protein Hydrolysate by a Mixture of Alcalase and Prozyme and Evaluation of Its Digestibility and Immunoregulatory Properties

  • Jiyeon Yang;Se Kyung Lee;Young Suk Kim;Hyung Joo Suh;Yejin Ahn
    • Food Science of Animal Resources
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    • v.43 no.4
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    • pp.594-611
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    • 2023
  • Whey protein (WP) has nutritional value, but the presence of β-lactoglobulin (β-LG) and α-lactalbumin (α-LA) cause allergic reactions. In this study, hypoallergenic whey protein hydrolyate (HWPH) was prepared by decomposing β-LG and α-LA of WP using exo- and endo-type proteases. The enzyme mixing ratio and reaction conditions were optimized using response surface methodology (RSM). Degradation of α-LA and β-LG was confirmed through gel electrophoresis, and digestion, and absorption rate, and immunostimulatory response were measured using in vitro and in vivo systems. Through RSM analysis, the optimal hydrolysis conditions for degradation of α-LA and β-LG included a 1:1 mixture of Alcalase and Prozyme reacted for 10 h at a 1.0% enzyme concentration relative to substrate. The molecular weight of HWPH was <5 kDa, and leucine was the prominent free amino acid. Both in vitro and in vivo tests showed that digestibility and intestinal permeability were higher in HWPH than in WP. In BALB/c mice, as compared to WP, HWPH reduced allergic reactions by inducing elevated Type 1/Type 2 helper T cell ratio in the blood, splenocytes, and small intestine. Thus, HWPH may be utilized in a variety of low allergenicity products intended for infants, adults, and the elderly.