• Title/Summary/Keyword: hybrid enzyme

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Antihypertensive Angiotensin I-Converting Enzyme Inhibitory Activity and Antioxidant Activity of Vitis hybrid-Vitis coignetiae Red Wine Made with Saccharomyces cerevisiae

  • Jang, Jeong-Hoon;Lee, Jong-Soo
    • Mycobiology
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    • v.39 no.2
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    • pp.137-139
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    • 2011
  • A Vitis hybrid-Vitis coignetiae red wine was vinified by fermentation of a mixture of a Vitis hybrid.Vitis coignetiae must with Saccharomyces cerevisiae KCTC 7904 at $25^{\circ}C$ for 10 days. The Vitis hybrid-Vitis coignetiae red wine showed high antihypertensive angiotensin I-converting enzyme (ACE) inhibitory activity (67.8%) and antioxidant activity (76.7%). The antihypertensive ACE inhibitor in the Vitis hybrid-Vitis coignetiae red wine was partially purified by solid phase extraction chromatography, and its ACE inhibitory activity yielded an $IC_{50}$ of 1.8 mg/mL. Six kinds of oligopeptides, including five new kinds, were contained in the partially purified ACE inhibitor fraction from the red wine after 10 days of fermentation. Antioxidant activity decreased significantly from 76.7% to 40.5% when the post-fermentation period was prolonged to 30 days.

Evaluation of the Estrogenic Activity by Yeast Two-hybrid Assay and Enzyme-linked Immunosorbent Assay in Sewage Treatment Plant (하수처리장의 내분비계장애물질에 대한 Yeast Two-hybrid Assay와 Enzyme-linked Immunosorbent Assay에 의한 에스트로겐활성도 평가)

  • Lee, Byoung-Cheun;Ra, Jin-Sung;Kim, Sang-Don;Kawai, Hukiko;Lee, Chul-Hee
    • Journal of Korean Society of Environmental Engineers
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    • v.29 no.7
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    • pp.771-777
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    • 2007
  • Several endocrine disrupting chemicals(EDCs) were monitored to evaluate the estrogenic activities and the concentrations by yeast two-hybrid assay and enzyme-linked immunosorbent assay(ELISA) in sewage treatment plant(STP) which consist of industrial and domestic line. In the influent of domestic line, estrone, 17$\beta$-estradiol, 17$\alpha$-ethinylestradiol and alkylphenolethoxylate(APE) were detected up to 167.1, 39.7, 7.3 and 145.4 ng/L, respectively. The average removal efficiency of 17$\beta$-estradiol after the activated sludge process was 77.5% and further removed to 80.8% after the sand filtration-ozonation step. These results suggests that the activated sludge process has limited potential to remove the estrogenic activity effectively. The contributions of the estrogenic chemicals to the estrogenic activities were 70.7, 23.3, 3.7 and 2.3% for estrone, 17$\beta$-estradiol 17$\alpha$-ethinylestradiol and APE, respectively, in the domestic line effluents. Therefore, 17$\beta$-estradiol and estrone contributed most of the estrogenic activity in the domestic line effluents.

Assessment of the Ability of Extracellular Enzyme Production in Hybrid Strains of Lentinula edodes by Chromogenic Reaction-based Plate Assay (발색반응 분석법을 이용한 표고 교배균주의 세포외효소 분비 능력 평가)

  • Kwon, Hyuk-Woo;Kim, Jun-Young;Ko, Han-Gyu;Park, Heung-Soo;Kim, Seong-Hwan
    • The Korean Journal of Mycology
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    • v.39 no.2
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    • pp.99-104
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    • 2011
  • Shiitake breeding requires the procedures of mating of two different parental strains and selection of hybrid strains that have good traits for the mushroom production. In this study, we tested the possibility of the use of chromogenic plate-based assay for extracellular enzyme production in order to assess and find good biochemical properties-possessed hybrid strains that were generated from genetic cross of the monokaryotic strains derived from two different parental strains of Lentinula edodes Sanjo-101ho and Sanjo-108ho. We observed that there was difference in the ability of producing ${\beta}$-glucosidase, avicelase, CM-cellulase, amylase, pectinase, xylanase, and protease among the monokaryotic strains. We could also comparatively assess that the ability of the seven extracellular enzymes production in the hybrid strains depended on the mating combination of the monokaryotic strains. Our results demonstrate that the assessment method for extracellular enzyme production using chromogenic plate assay could be usefully applied to the assessment of the hybrid strains derived from the breeding procedure of L. edodes.

Chemical Synthesis and Cloning of Panax ginseng Peptide Gene

  • Zhang, Hong-Ying;Chen, Dong-Song;Zhang, Jin
    • Proceedings of the Ginseng society Conference
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    • 1990.06a
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    • pp.65-67
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    • 1990
  • The sequence of ginseng peptide gene was designed and synthesized by the solid phase plasmid pUC19. Escherichia coli JM101 cells were transformed with above hybrid plasmids. Ampicillin resistant transformants were screened and identified by in situ colony hybridization and Southern blot techinques. Finally the gene sequencing was done by the Sanger dideoxy method using primer extension.

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Chemical Synthesis and Cloning of Panax ginseng Peptide Gene (인삼펩티드 유전자의 합성 및 클로닝)

  • Zhang, Hong-Ying;Chen, Dong-Song;Zhang, Jin
    • Journal of Ginseng Research
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    • v.14 no.2
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    • pp.207-209
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    • 1990
  • The sequence of ginseng peptide gene was designed and synthesized by the solid phase phosphoramidiate method. Synthetic segments were isolated, pllrified and joined to the plasmid pUC19. E.icherichiu coli JM101 cells were transformed with above hybrid plasmids. AmpiciIBin resistant transformants were screened and identified by in situ colony hybridization and Southern blot techniques. Finally the gene sequencing was done by the Sanger dideoxy method using primer extension.

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An organofunctionalized MgO∙SiO2 hybrid support and its performance in the immobilization of lipase from Candida rugosa

  • Kolodziejczak-Radzimska, Agnieszka;Zdarta, Jakub;Ciesielczyk, Filip;Jesionowski, Teofil
    • Korean Journal of Chemical Engineering
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    • v.35 no.11
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    • pp.2220-2231
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    • 2018
  • Lipase from Candida rugosa was immobilized on $MgO{\cdot}SiO_2$ hybrid grafted with amine, thiol, cyano, phenyl, epoxy and carbonyl groups. The products were analyzed using Fourier transform infrared spectroscopy, nuclear magnetic resonance, low-temperature $N_2$ sorption and elemental analysis. Additionally, the degree of coverage of the oxide material surface with different functional groups and the number of surface functional groups were estimated. The Bradford method was used to determine the quantity of immobilized enzyme. The largest quantity of enzyme (25-28 mg/g) was immobilized on the hybrid functionalized with amine and carbonyl groups. On the basis of hydrolysis reaction of p-nitrophenyl palmitate to p-nitrophenol, it was determined how the catalytic activity of the obtained biocatalysts is affected by pH, temperature, storage time, and repeated reaction cycles. The best results for catalytic activity were obtained for the lipase immobilized on $MgO{\cdot}SiO_2$ hybrids with amine and carbonyl groups. The biocatalytic system demonstrated activity above 40% in the pH range 4-10 and in the temperature range $30-70^{\circ}C$. Lipase immobilized on the $MgO{\cdot}SiO_2$ systems with amine and epoxy groups retains, respectively, around 80% and 60% of its initial activity after 30 days of storage, and approximately 60-70% after 10 reaction cycles.

Purification of carbosymethyl cellulase from hybrid between aspergillus niger and penicillium verruculosum

  • Yang, Young-Ki;Lee, Jung-Sup;Park, Hyung-Nam;Moon, Myung-Nim;Kim, Hong-Sub;Kim, Jong-Se;Lim, Chae-Young;Rhee, Young-Ha
    • Journal of Microbiology
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    • v.34 no.1
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    • pp.90-94
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    • 1996
  • The carboxymethyl cellulase (CMCase) was purified from the induced culture filtrate of hybrid TAPW15703 between Aspergillus niger and penicillium verruculosum made by nuclear transfer. The enzyme was purified 80 fold with an overall yield 17% from the culture medium by ammonium sulfate fractionation, Sephadex G-75 gel permeation chromatography, and DEAE-ion exchange column chromatography. The molecular weight of the CMCase has estimated to be 32,000 daltons on SDS-polyacrylamide gel electrophoresis and Sephadex G-150 gel permeation chromatography. The purified enzyme functions optimally at pH 4.0 and 4$0^{\circ}C$ The Km value for carbosymethyl cellulose was 68 mM. The enzyme activity was increased by the presence of $Mg^{2+}$and Mn$^{2+}$.

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Immobilization of Xylanase Using a Protein-Inorganic Hybrid System

  • Kumar, Ashok;Patel, Sanjay K.S.;Mardan, Bharat;Pagolu, Raviteja;Lestari, Rowina;Jeong, Seong-Hoon;Kim, Taedoo;Haw, Jung Rim;Kim, Sang-Yong;Kim, In-Won;Lee, Jung-Kul
    • Journal of Microbiology and Biotechnology
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    • v.28 no.4
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    • pp.638-644
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    • 2018
  • In this study, the immobilization of xylanase using a protein-inorganic hybrid nanoflower system was assessed to improve the enzyme properties. The synthesis of hybrid xylanase nanoflowers was very effective at $4^{\circ}C$ for 72 h, using 0.25 mg/ml protein, and efficient immobilization of xylanase was observed, with a maximum encapsulation yield and relative activity of 78.5% and 148%, respectively. Immobilized xylanase showed high residual activity at broad pH and temperature ranges. Using birchwood xylan as a substrate, the $V_{max}$ and $K_m$ values of xylanase nanoflowers were 1.60 mg/ml and $455{\mu}mol/min/mg$ protein, compared with 1.42 mg/ml and $300{\mu}mol/min/mg$ protein, respectively, for the free enzyme. After 5 and 10 cycles of reuse, the xylanase nanoflowers retained 87.5% and 75.8% residual activity, respectively. These results demonstrate that xylanase immobilization using a proteininorganic hybrid nanoflower system is an effective approach for its potential biotechnological applications.

Changes of Angiotensin I-Converting Enzyme Inhibitory Activity, Fibrinolytic Activity and $\beta$-Secretase Inhibitory Activity of Red Wines During Fermentation and Post-Fermentation (적포도주들의 발효와 후 발효 중 심혈관 관련 Angiotensin I 전환효소 저해활성과 혈전용해활성 및 $\beta$-secretase 저해 활성의 변화)

  • No, Jae-Duck;Lee, Eun-Na;Seo, Dong-Soo;Chun, Jong-Pil;Choi, Shin-Yang;Lee, Jong-Soo
    • Microbiology and Biotechnology Letters
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    • v.36 no.4
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    • pp.291-298
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    • 2008
  • The cardiovascular angiotensin I-converting enzyme inhibitory activity, fibrinolytic activity and bbb-secretase inhibitory activity of four kinds of red wine were investigated during fermentation and post-fermentation. After 10 days of fermentation, the antihypertensive angiotensin I-converting enzyme (ACE) inhibitory activities of all the red wines ranged from 38.6% to 58.8%. However, the ACE inhibitory activities increased with the prolongation of the post-fermentation period; moreover, in the Vitis hybrid red wine, the ACE inhibitory activity reached its highest value, 76.9%, after 120 days of post-fermentation. During the fermentation and post-fermentation of all the red wines, fibrinolytic activity was weak or not detected. After 10 days of fermentation, Vitis labrusca B red wine exhibited the greatest antidementia $\beta$-secretase inhibitory activity of 54.8%, though $\beta$-secretase inhibitory activity decreased significantly to less than 10% during 120 days of post-fermentation. In conclusion, we obtained a highly valuable Vitis hybrid red wine that was fermented for 10 days at $25^{\circ}C$ with Vitis hybrid and S. cerevisiae K-7 and then post-fermentation for 120 days at $4^{\circ}C$.