• Title/Summary/Keyword: acid-fusion

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Cloning and Prokaryotic Expression of C-type Lysozyme Gene from Agrius convolvuli

  • Kim, Jong-Wan;Yoe, Sung-Moon
    • Animal cells and systems
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    • v.12 no.3
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    • pp.149-155
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    • 2008
  • We have isolated and characterized Agrius convolvuli cDNA encoding a c-type lysozyme. The cDNA sequence encodes a processed protein of 139 amino acid residues with 19 amino acid residues amino-terminal signal sequence and 120 amino acid residues mature sequence. The amino acid residues responsible for the catalytic activity and the binding of the substrate are conserved. Agrius lysozyme has a high identity to Manduca sexta. Recombinant A. convolvuli lysozyme was expressed in Escherichia coli BL21(DE3) pLysS cells for pGEX 4T-1 expression vector. Their optimal conditions for the fusion protein expression and purification were screened. Lysozyme gene amplified with primers ACLyz BamHI and ACLyz XhoI was ligated into the pGEX 4T-1 vector, which contained the glutathione S-transferase(GST) gene for fusion partner. The fusion protein was induced by IPTG and identified by SDS-PAGE analysis. Molecular weight of the fusion protein was estimated to be about 45 kDa. Recombinant lysozyme, fused to GST, was purified by glutathion-Sepharose 4B affinity chromatography. Western blot analysis of this protein revealed an immunoreactivity with the anti-Agrius lysozyme.

Studies of the Fusibility of Coal Ashes in Oxidizing and Reducing Conditions (산화성 및 환원성분위기에서 석탄회분의 용융성)

  • Park, Chu-Sik;Lee, Shi-Hun;Choi, Sang-Il;Yang, Hyun-Soo
    • Applied Chemistry for Engineering
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    • v.8 no.2
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    • pp.179-190
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    • 1997
  • To study the effects of chemical composition on the fusion temperatures of coal ashes, the chemical composition, mineral matter, and fusion temperature were studied with 54 kinds of coal ash samples including Korean anthracite coals. CaO, MgO and $Fe_2O_3$ were observed to be major fluxing elements in reducing and oxidizing atmosphere. The fluxing effect of $Fe_2O_3$ was increased more in reducing atmosphere. In a base/acid ratio, the fusion temperature decreased with increasing amounts of basic components. Nevertheless, the correlation between a fusion temperature and base/acid ratio was not shown well in a higher ratio of $Fe_2O_3/CaO$. The differences of fusion temperatures between oxidizing and reducing atmosphere showed close relationship with $SiO_2/Al_2O_3$ ratio rather than with $Fe_2O_3$ contents. Multiple regression was used to predict the fusion temperature of coal ashes, and it was established that the major predictors in oxidizing atmosphere were Base/Acid, $Fe_2O_3/CaO$, $SiO_2/Al_2O_3$, and $(SiO_2/A1_2O_3){\cdot}(Base/Acid)$ and Base/Acid, $Fe_2O_3/CaO$, $SiO_2$, and $TiO_2$ were major ones in reducing atmosphere.

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Regeneration of Intervertebral Disc Using Poly(lactic-co-glycolic acid) Scaffolds Included Demineralized Bone Particle In Vivo (In vivo 상에서 탈미네랄화된 골분이 함유된 PLGA 지지체를 이용한 추간판 디스크 재생)

  • Jang, Ji Eun;Kim, Hye Yoon;Song, Jeong Eun;Lee, Dongwon;Kwon, Soon Yong;Chung, Jin Wha;Khang, Gilson
    • Polymer(Korea)
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    • v.37 no.6
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    • pp.669-676
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    • 2013
  • Demineralized bone particle (DBP) is a biomaterial used widely in the field of tissue engineering. In this study, in order to study the effect of DBP/poly(lactic-co-glycolic acid) (PLGA) scaffold on disc regeneration in vivo environment, we prepared the porous DBP/PLGA hybrid scaffold. Disc defect was induced by removing the nucleus pulposus tissue after incision the annulus fibrosus tissue in half and scaffolds were transplanted. After 1, 2 and 3 months later, the extracted discs were confirmed by collagen synthesis and glycosaminoglycan (sGAG). We conducted histology (H&E, Safranin-O, Alcian blue, Type I Collagen, Type II Collagen). From the results, it was confirmed that collagen and sGAG content were high in DBP/PLGA scaffold, and the regeneration of intervertebral disc was possible.

Protoplast Fusion Between Aspergillus oryzae and Aspergillus shirousamii (Aspergillus oryzae와 Aspergillus shirousamii간의 원형질체의 융합)

  • Shin, Dong-Bun;Ryu, Beung-Ho;Jin, Seung-Heun
    • Korean Journal of Food Science and Technology
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    • v.25 no.4
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    • pp.366-372
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    • 1993
  • This study mainly designed to high quality of mirin production by using protopast fusion. In order to enhance the acid carboxypeptidase (ACPase) activity by the method of protoplast fusion. In order to enhance the acid carboxypeptidase (ACPase) activity by the method of protopalst fusion, the mutants, Aspergillus oryzae 9-12 and Aspergillus shirosamii IFO 6082-60 were selected by mutation among various mutants. Protoplast of Aspergillus oryzae 9-12 and Aspergillus shirousamii IFO 6082-60 were formed effectively by incubation of the mixtures of chitinase (10mg/ml), cellulase (10mg/ml) and zymolase 20T (5mg/ml). For protopalst fusion, the mixture of two mutant were fused to effective under the optimum conditions by solutions containing 30% PEG 6,000, 0.01M $CaCl_2\;2H_2O$, 0.6M KCl and 0.05M glycine. Fusion frequency was 0.71% and fusant, F-50 appeared ACPase activity of 20,800 unit/g which has 1.5 times higher than that of each mutants.

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Effects of comonomer with carboxylic group on stabilization of high molecular weight polyacrylonitrile nanofibrous copolymers

  • Lei, Danyun;Devarayan, Kesavan;Li, Xiang-Dan;Choi, Woong-Ki;Seo, Min-Kang;Kim, Byoung-Suhk
    • Carbon letters
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    • v.15 no.4
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    • pp.290-294
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    • 2014
  • New precursors, poly(acrylonitrile-co-crotonic acid) (poly(AN-CA)) and poly(acrylonitrile-co-itaconic acid-co-crotonic acid) (poly(AN-IA-CA)) copolymers, for the preparation of carbon fibers, were explored in this study. The effects of comonomers with acidic groups, such as crotonic acid (CA) and/or itaconic acid (IA), on the stabilization of nanofibrous polyacrylonitrile (PAN) copolymers were studied. The extent of stabilization, evaluated by Fourier transform infrared spectroscopy, revealed that the CA comonomer could retard/control the stabilization rate of PAN, in contrast to the IA comonomer, which accelerated the stabilization process. Moreover, the synthesized PAN copolymers containing CA possessed higher Mv than those of the IA copolymers and also showed outstanding dimension stability of nanofibers during the stabilization, which may be a useful property for improving the dimensional stability of polymer composites during manufacturing.

Effects of Demineralized Bone Particle Loaded Poly(lactic-co-glycolic acid) Scaffolds on the Attachment and Proliferation of Costal Cartilage Cells (탈미네랄화된 골분/PLGA 지지체에서 늑연골 세포의 부착과 성장에 미치는 영향)

  • Cho, Sun Ah;Song, Jeong Eun;Kim, Kyoung Hee;Ko, Hyun Ah;Lee, Dongwon;Kwon, Soon Yong;Chung, Jin Wha;Khang, Gilson
    • Polymer(Korea)
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    • v.37 no.5
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    • pp.632-637
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    • 2013
  • It has been widely accepted that costal cartilage cells (CCs) have more excellent initial proliferation capacity than articular cartilage cells as well as the easiness for isolation and collection. This study demonstrated that CCs might be one of the substitutes for articular cartilage cells by tissue engineered cartilage. Poly(lactic-co-glycolic acid) (PLGA) has been extensively tested and used as scaffold material but it was limited by the low attachment of cells and the induction of inflammatory cells. Base on previous our studies, we confirmed demineralized bone particle (DBP) had the power of the reduction of inflammatory reaction and the stimulation proliferation of cells. We fabricated PLGA scaffold loaded with 10, 20, 40 and 80 wt% DBP and then tested the possibility of the regeneration of cartilage using CCs. Assays of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and scanning electron microscope (SEM) carried out to evaluate the attachment and proliferation of CCs in DBP/PLGA scaffolds. Glycosaminoglycan (sGAG) and collagen contents assay were conducted to confirm the effects of DBP on formation of extracellular matrix. This study demonstrated that DBP/PLGA scaffolds showed significant positive effects on cell growth and proliferation due to the vitality of DBP as well as the possibility of the application of CCs for tissue engineered cartilage.

Chemometric Approach to Fatty Acid Profiles in Soybean Cultivars by Principal Component Analysis (PCA)

  • Shin, Eui-Cheol;Hwang, Chung-Eun;Lee, Byong-Won;Kim, Hyun-Tae;Ko, Jong-Min;Baek, In-Youl;Lee, Yang-Bong;Choi, Jin-Sang;Cho, Eun-Ju;Seo, Weon-Taek;Cho, Kye-Man
    • Preventive Nutrition and Food Science
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    • v.17 no.3
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    • pp.184-191
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    • 2012
  • The purpose of this study was to investigate the fatty acid profiles in 18 soybean cultivars grown in Korea. A total of eleven fatty acids were identified in the sample set, which was comprised of myristic (C14:0), palmitic (C16:0), palmitoleic (C16:1, ${\omega}7$), stearic (C18:0), oleic (C18:1, ${\omega}9$), linoleic (C18:2, ${\omega}6$), linolenic (C18:3, ${\omega}3$), arachidic (C20:0), gondoic (C20:1, ${\omega}9$), behenic (C22:0), and lignoceric (C24:0) acids by gas-liquid chromatography with flame ionization detector (GC-FID). Based on their color, yellow-, black-, brown-, and green-colored cultivars were denoted. Correlation coefficients (r) between the nine major fatty acids identified (two trace fatty acids, myristic and palmitoleic, were not included in the study) were generated and revealed an inverse association between oleic and linoleic acids (r=-0.94, p<0.05), while stearic acid was positively correlated to arachidic acid (r=0.72, p<0.05). Principal component analysis (PCA) of the fatty acid data yielded four significant principal components (PCs; i.e., eigenvalues>1), which together account for 81.49% of the total variance in the data set; with PC1 contributing 28.16% of the total. Eigen analysis of the correlation matrix loadings of the four significant PCs revealed that PC1 was mainly contributed to by oleic, linoleic, and gondoic acids, PC2 by stearic, linolenic and arachidic acids, PC3 by behenic and lignoceric acids, and PC4 by palmitic acid. The score plots generated between PC1-PC2 and PC3-PC4 segregated soybean cultivars based on fatty acid composition.

Intraspecific Protoplast Fusion of Citric Acid Producer, Candida lipolytica (구연산 생성 Candida lipolytica의 원형질체 융합)

  • 성낙계;심기환;전효곤;강신권;박석규
    • Microbiology and Biotechnology Letters
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    • v.13 no.4
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    • pp.391-395
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    • 1985
  • In order to develope a protoplast fusion system for citric acid and SCP producing Candida lipolytica, the optimal conditions for the formation and regeneration of protoplast were examined and the protoplast fusion was performed. At the optimal conditions of growth phase and Zymolyase treatment, frequencies of protoplast formation were 98%. Approximately 20-30% of protoplasts were regenerated on the regeneration minimal medium containing 3% agar and 30mM $CaCl_2$ with the overlay of the same medium. The fusion frequencies, 4-5${\pm}$10$^{-4}$, were accomplished by the treatment of two nutritionally complementary auxotrophic protoplasts, L-14 ($lys^-$) and T-24 (X$30^-$), with 30% PEG 6000 containing 100mM $CaCl_2$ at $30^{\circ}C$ for 20 minutes.

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