• Title/Summary/Keyword: thermal and pH stability

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Purification of xylose reductase from Candida sp. BT001 and characterization of its properties (Candida sp. BT001의 xylose reductase의 정제 및 성질)

  • Hwang, In-Gyun;Lee, Sang-Hyub;Lee, Wang-Sik;Bang, Won-Gi
    • Applied Biological Chemistry
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    • v.36 no.3
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    • pp.178-183
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    • 1993
  • Xylose reductase (alditol: $NADP^+$ 1-oxidoreductase, EC 1.1.1.21) from the xylose-fermenting yeast, Candida sp. BT001, was purified via salt fractionation, ion-exchange, gel filtration and affinity chromatography, and its properties were characterized. The enzyme from the yeast was active with both NADPH and NADH as coenzyme. The xylose reductase activity with NADH was approximately 51% of that with NADPH and the specific activities of purified enzyme with NADPH and NADH were 11.78 U/mg and 6.01 U/mg, respectively. Molecular weight of the purified enzyme was 31,000 on SDS-PAGE and 61,000 on gel filtration. The Km for D-xylose, NADPH, and NADH was $94.2{\times}10^{-3}M,\;0.011{\times}10^{-3}M\;and \;0.032{\times}10^{-3}M$, respectively. The purified xylose reductase had relatively higher substrate affinity for L-arabinose than other aldoses tested. The optimal pH was 6.2 and the optimal reaction temperature was $45^{\circ}C$. The thermal stability of the enzyme was for 20 minutes at $30^{\circ}C$.

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Characterization of Transparent TiO2 Power and Thin Films through Sol-Gel Process (졸-겔법을 이용한 투과성 TiO2 분말 및 박막의 특성 연구)

  • Jung, Mie-Won;Lee, Zee-Young;Son, Hyun-Jin
    • Journal of the Korean Ceramic Society
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    • v.39 no.3
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    • pp.252-258
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    • 2002
  • Transparent $TiO_2$ thin films prepared by sol-gel process using the modification of titanium(IV) alkoxide showed improved thermal stability and high refraction index. Compared to the pure $TiO_2$ film, the modified $TiO_2$ films show the increased index of refraction under proper condition at pH 2.5. Transparency of these $TiO_2$ thin films were more than 80% in the visible region. It has been demonstrated that the reaction occurs in the amorphous phase: an exchange of phase results in anatase before and after 400$^{\circ}C$, in rutile over 700$^{\circ}$C form the XRD results. The particle sizes, shapes and structures of these nanoclusters in the $TiO_2$ films have been identified through a SEM and XRD. The physical properties and structures of their powders have also been studied through a SEM, XRD, TGA and DSC. The thickness and index of refraction have been determined by the analysis of ellipsometric spectra.

The effect of antagonists produced by Paenibacillus polymyxa CK-1 on the growth of Trichoderma sp. (Paenibacillus polymyxa CK-1이 생산한 길항물질이 Trichoderma sp. 생육에 미치는 영향)

  • Lee, Sang-Won;Choi, Jin-Sang;Kim, Chul-Ho
    • Journal of Mushroom
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    • v.12 no.3
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    • pp.201-208
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    • 2014
  • The separation of the bacteria inhibiting Trichoderma sp. mold, the strain causing blue mold disease that occurs frequently when cultivating mushroom while carrying out the efficient fermentation of mushroom medium, from the growth was done. In about 200 strains isolated primarily from fungus garden samples, 6 strains were secondly isolated, which had fast growth rates and a clear zone on the plate medium of SM, AM, and CM. Among the 6 strains isolated, the C-1 strain showed high enzymatic activity of cellulase, amylase, and protease, and strong antibacterial activity for the T. virens and T. harzianum, selected finally. The selected C-1 strain was identified as Paenibacillus polymyxaby the result of the identification by Bergey's Manual of Systematic Bacteriology and the analysis of the nucleotide sequence of 16S rRNA, and named as P. polymyxa CK-1. In reviewing the growth conditions of the P. polymyxa CK-1 strain, the optimum cultivation temperature was $45^{\circ}C$, and the optimum pH for growth was in the range of 6.0~7.0. Appropriate incubation time of P. polymyxa CK-1 for the growth inhibition of the fungus T. virens and T. harzianum was 22 to 36 hours. And the fungal growth was not observed, even when leaving two molds inoculated on each petri dishes, which were treated with 24 hour culture solution of P. polymyxa CK-1 strain for 10 days. As a result of studying the thermal stability of the antagonists produced by the P. polymyxa CK-1 strain, no mycelial growth of the two fungi was observed in the test group treated for 20 minutes at $60^{\circ}C$ and $100^{\circ}C$, but mycelial growth was slightly observed in the test group treated for 20 minutes at $121^{\circ}C$. As aresult of reviewing the impact of the P. polymyxa CK-1 culture medium on mushroom mycelial growth, it showed no effect on a variety of mushroom mycelial growth including enoki mushroom and shiitake mushroom.

Preparation of Protein-coated Cationic Liposomes Containing Doxorubicin and Their Binding Property of Blood Plasma Protein (독소루비신을 함유하고 단백질로 수식된 양이온성 리포솜의 제조 및 혈장 단백흡착 특성)

  • Kim, Sung-Kyu;Jung, Soon-Hwa;Jung, Suk-Hyun;Seong, Ha-Soo;Chi, Sang-Cheol;Cho, Sun-Hang;Shin, Byung-Cheol
    • Journal of the Korean Chemical Society
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    • v.52 no.1
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    • pp.57-65
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    • 2008
  • are nanometer or micrometer scale vesicles that can be used as drug delivery carriers. However, plain liposomes are plagued by rapid opsonization, making their circulation time in bloodstream be shortened. In this study, model protein, bovine serum albumin (BSA)-coated liposomes were prepared by coating cationic liposomes with BSA molecules at higher pH than isoelectric point of BSA. The BSA molecules coated on the liposomal surface were denatured by thermal treatment at above 60oC. While both plain and cationic liposomes had about mean particle diameter of 1041 nm, BSA-coated cationic liposomes (BCL) had mean particle diameter of 1091 nm. Encapsulation of model drug, doxorubicin (DOX), in liposomes were carried out by using remote loading method and the loading efficiency of DOX to liposomes was about 90%. The mean particle diameter of BCL did not increase in blood plasma and adsorption of plasma protein was much less than plain or cationic liposomes. These results suggest that BCL can be used as a long-circulating liposomes in bloodstream.

Prevention of Discoloration and Storage Stability in Canned Ark Shell (새고막 통조림 변색방지 및 저장중 품질변화)

  • 배태진;김귀식
    • The Korean Journal of Food And Nutrition
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    • v.11 no.2
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    • pp.243-248
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    • 1998
  • Ark shell was known as shellfish that had hemoglobin as blood pigment and the action of mecidine, was consumed the great part of it as raw material, though it was produced about 13,000 M/T per year. Ark shell was processed the infinitesimal quantity as conned product, bout canned ark shell had problem that occurrenced discoloration after heat treatment during processing and storage. This discoloration mechanism during processing and storage was not cleared. This study was carried out to understand characteristics of the hemoglobin as blood pigment and carotenoid as meat pigment in ark shell and management of proper processing conditions for prevention of oxidation and discoloration by thermal treatment. When treated by digestion of 0.1% BHA, 0.1% Tenox-II, 0.5% Na2EDTA, 0.05% NDGA and 3% salt soln., 0.1% BHA solution was most suitable for stability of carotenoid that the retention ratio of carotenoids were 63.1% after heating to 116$^{\circ}C$ for 120 minutes. In preparation of canned ark shell and storage at 37$\pm$1$^{\circ}C$ for 60 days, the chemical composition, pH and salinity ere stable. And contents of total carotenoid were decreased slightly from 0.83mg% to 0.727mg%. The viable cell count were 6.92$\times$103 cfu/ml at raw ark shell, after processed and storage were not detected. The predominant amino acids in the raw ark shell were glutamic acid(19.7%), arginine(16.0%), glycine(12.6%), alanine(12.2%) and aspartic acid(7.6%). When 60 days stored, the contents of amino acid were stable. And the predominant nuclotide and their related compounds in the raw ark shell were hypoxanthine(2.14$\mu$mol/g), IMP(1.94$\mu$mol/g) and ATP(0.87$\mu$mol/g), and storage at 37$\pm$1$^{\circ}C$ for 60 days, the quantity order were same as raw material.

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Improving the Chitinolytic Activity of Bacillus pumilus SG2 by Random Mutagenesis

  • Vahed, Majid;Motalebi, Ebrahim;Rigi, Garshasb;Noghabi, Kambiz Akbari;Soudi, Mohammad Reza;Sadeghi, Mehdi;Ahmadian, Gholamreza
    • Journal of Microbiology and Biotechnology
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    • v.23 no.11
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    • pp.1519-1528
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    • 2013
  • Bacillus pumilus SG2, a halotolerant strain, expresses two major chitinases designated ChiS and ChiL that were induced by chitin and secreted into the supernatant. The present work aimed to obtain a mutant with higher chitinolytic activity through mutagenesis of Bacillus pumilus SG2 using a combination of UV irradiation and nitrous acid treatment. Following mutagenesis and screening on chitin agar and subsequent formation of halos, the mutated strains were examined for degradation of chitin under different conditions. A mutant designated AV2-9 was selected owing to its higher chitinase activity. To search for possible mutations in the whole operon including ChiS and ChiL, the entire chitinase operon, including the intergenic region, promoter, and two areas corresponding to the ChiS and ChiL ORF, was suquenced. Nucleotide sequence analysis of the complete chitinase operon from the SG2 and AV2-9 strains showed the presence of a mutation in the catalytic domain (GH18) of chitinase (ChiL). The results demonstrated that a single base change had occurred in the ChiL sequence in AV2-9. The wild-type chitinase, ChiL, and the mutant (designated ChiLm) were cloned, expressed, and purified in E. coli. Both enzymes showed similar profiles of activity at different ranges of pH, NaCl concentration, and temperature, but the mutant enzyme showed approximately 30% higher catalytic activity under all the conditions tested. The results obtained in this study showed that the thermal stability of chitinase increased in the mutant strain. Bioinformatics analysis was performed to predict changes in the stability of proteins caused by mutation.

Combustion Characteristic Study of LNG Flame in an Oxygen Enriched Environment (산소부화 조건에 따른 LNG 연소특성 연구)

  • Kim, Hey-Suk;Shin, Mi-Soo;Jang, Dong-Soon;Lee, Dae-Geun
    • Journal of Korean Society of Environmental Engineers
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    • v.29 no.1
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    • pp.23-30
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    • 2007
  • The ultimate objective of this study is to develop oxygen-enriched combustion techniques applicable to the system of practical industrial boiler. To this end the combustion characteristics of lab-scale LNG combustor were investigated as a first step using the method of numerical simulation by analyzing the flame characteristics and pollutant emission behaviour as a function of oxygen enrichment level. Several useful conclusions could be drawn based on this study. First of all, the increase of oxygen enrichment level instead of air caused long and thin flame called laminar flame feature. This was in good agreement with experimental results appeared in open literature and explained by the effect of the decrease of turbulent mixing due to the decrease of absolute amount of oxidizer flow rate by the absence of the nitrogen species. Further, as expected, oxygen enrichment increased the flame temperatures to a significant level together with concentrations of $CO_2$ and $H_2O$ species because of the elimination of the heat sink and dilution effects by the presence of $N_2$ inert gas. However, the increased flame temperature with $O_2$ enriched air showed the high possibility of the generation of thermal $NO_x$ if nitrogen species were present. In order to remedy the problem caused by the oxygen-enriched combustion, the appropriate amount of recirculation $CO_2$ gas was desirable to enhance the turbulent mixing and thereby flame stability and further optimum determination of operational conditions were necessary. For example, the adjustment of burner with swirl angle of $30\sim45^{\circ}$ increased the combustion efficiency of LNG fuel and simultaneously dropped the $NO_x$ formation.

Studies on the Processings of Sterilized Salt-Fermented Anchovy Sauces (멸치액젓의 레토르트 식품화에 관한 연구)

  • Oh, Kwang-Soo
    • Korean Journal of Food Science and Technology
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    • v.28 no.6
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    • pp.1038-1044
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    • 1996
  • The salt-fermented anchovy sauce (AS) was packed in round No. 307-1 can, and thermally processed at $121.1^{\circ}C$ to obtain Fo values of 3, 5 and 10. The changes of food components and qualifies by thermal processing of sterilized AS (RAS) were examined. The compositions of AS were as followed; pH 6.81, VBN 394.7 mg/100g, total nitrogen 2,195.5 mg%, amino-nitrogen 1,010.5 mg%, and acidity 10.5 ml. Viable cell counts of AS on 0%, 5%, 20% NaCl-medium were $2.9{\times}10^3,\;9.1{\times}10^3$ and not detected, respectively. And viable cell counts of RAS were not detected. Acidity, total nitrogen and amino nitrogen contents of AS decreased slightly with the severeness of heat treatments, whereas pH and VBN content were increased. Total free amino acid contents of raw AS and RAS were 12,802.5 mg% and $11,212.6{\sim}12,105.4\;mg%$, and major amino acids were alanine, glutamic acid, leucine, isoleucine, valine and lysine. Also contents of IMP, hypoxanthine, TMAO and TMA in AS and RAS were 42.1 mg% and $35.5{\sim}40.9\;mg%$, 103.7 mg% and $103.1{\sim}105.5\;mg%$, 78.8 mg% and $58.2{\sim}71.6\;mg%$, 55.8 mg% and $58.9{\sim}68.5\;mg%$ respectively. And a little changes were observed in whole volatile components of AS with severeness of heat treatments by GC chromatogram patterns. Judging from the chemical and sensory evaluations, the Fo 3 heat treatment sample was not inferior to raw AS, and maintained good quality for 1 year storage.

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Cloning, Heterologous Expression, and Characterization of Novel Protease-Resistant ${\alpha}$-Galactosidase from New Sphingomonas Strain

  • Zhou, Junpei;Dong, Yanyan;Li, Junjun;Zhang, Rui;Tang, Xianghua;Mu, Yuelin;Xu, Bo;Wu, Qian;Huang, Zunxi
    • Journal of Microbiology and Biotechnology
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    • v.22 no.11
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    • pp.1532-1539
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    • 2012
  • The ${\alpha}$-galactosidase-coding gene agaAJB13 was cloned from Sphingomonas sp. JB13 showing 16S rDNA (1,343 bp) identities of ${\leq}97.2%$ with other identified Sphingomonas strains. agaAJB13 (2,217 bp; 64.9% GC content) encodes a 738-residue polypeptide (AgaAJB13) with a calculated mass of 82.3 kDa. AgaAJB13 showed the highest identity of 61.4% with the putative glycosyl hydrolase family 36 ${\alpha}$-galactosidase from Granulicella mallensis MP5ACTX8 (EFI56085). AgaAJB13 also showed <37% identities with reported protease-resistant or Sphingomonas ${\alpha}$-galactosidases. A sequence analysis revealed different catalytic motifs between reported Sphingomonas ${\alpha}$-galactosidases (KXD and RXXXD) and AgaAJB13 (KWD and SDXXDXXXR). Recombinant AgaAJB13 (rAgaAJB13) was expressed in Escherichia coli BL21 (DE3). The purified rAgaAJB13 was characterized using p-nitrophenyl-${\alpha}$-D-galactopyranoside as the substrate and showed an apparent optimum at pH 5.0 and $60^{\circ}C$ and strong resistance to trypsin and proteinase K digestion. Compared with reported proteaseresistant ${\alpha}$-galactosidases showing thermolability at $50^{\circ}C$ or $60^{\circ}C$ and specific activities of <71 U/mg with or without protease treatments, rAgaAJB13 exhibited a better thermal stability (half-life of >60 min at $60^{\circ}C$) and higher specific activities (225.0-256.5 U/mg). These sequence and enzymatic properties suggest AgaAJB13 is the first identified and characterized Sphingomonas ${\alpha}$-galactosidase, and shows novel protease resistance with a potential value for basic research and industrial applications.

Partial Oxidation of Methane to $H_2$ Over Pd/Ti-SPK and Pd/Zr-SPK Catalysts and Characterization (Pd/Ti-SPK과 Pd/Zr-SPK 촉매상에서 수소 생산을 위한 메탄의 부분산화반응과 촉매의 특성화)

  • Seo, Ho-Joon;Kang, Ung-Il
    • Applied Chemistry for Engineering
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    • v.21 no.6
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    • pp.648-652
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    • 2010
  • Catalytic activities of the partial oxidation of methane (POM) to hydrogen were investigated over Pd(5)/Ti-SPK and Pd(5)/Zr-SPK in a fixed bed flow reactor (FBFR) under atmosphere, and the catalysts were characterized by BET, XPS, XRD. The BET surface areas, pore volume and pore width of Horvath-Kawaze, micro pore area and volume of t-plot of Pd(5)/Ti-SPK and Pd(5)/Zr-SPK were $284m^2/g$, $0.233cm^3/g$, 3.9 nm, $30m^2/g$, $0.015cm^3/g$ and $396m^2/g$, $0.324cm^3/g$, 3.7nm, $119m^2/g$, $0.055cm^3/g$, repectively. The nitrogen adsorption isotherms were type IV with hysteresis. XPS showed that Si 2p and O 1s core electronlevels of Ti-SPK and Zr-SPK substituted Ti and Zr shifted to slightly lower binding energies than SPK. The oxidation states of Pd on the surface of catalysts were $Pd^0$ and $Pd^{+2}$. XRD patterns showed that crystal structures of fresh catalyst changed amorphous into crystal phase after reaction. The conversion and selectivity of POM to hydrogen over Pd(5)/Ti-SPK and Pd(5)/Zr-SPK were 77, 84% and 78, 72%, respectively, at 973 K, $CH_4/O_2$ = 2, GHSV = $8.4{\times}10^4mL/g_{cat}{\cdot}h$ and were kept constant even after 3 days in stream. These results confirm superior activity, thermal stability, and physicochemical properties of catalyst in POM to hydrogen.