• Title/Summary/Keyword: enzymatic characterization

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Mycoflora and Enzymatic Characterization of Fungal Isolates in Commercial Meju, Starter for a Korean Traditional Fermented Soybean Product

  • Baek, Jin-Ho;So, Kum-Kang;Ko, Yo-Han;Kim, Jung-Mi;Kim, Dae-Hyuk
    • Mycobiology
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    • v.42 no.3
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    • pp.291-295
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    • 2014
  • Mycoflora was assessed in the commercial meju from four well-separated geographic origins. A total of 112 fungal isolates were identified by phenotypic characteristics and molecular taxonomy using sequencing the internal transcribed spacer of the rDNA and revealed 19 species from 13 genera. Enzymatic characteristics of protease and amylase, and mycotoxin production were analyzed.

Enzymatic sccharification of lignocellulosic biomass by enzyme system of brown-rot fungi (갈색부후균의 효소시스템을 이용한 목질계 바이오매스의 효소당화)

  • Yoon, Jeong-Jun;Cha, Chang-Jun;Kim, Yeong-Suk;Kim, Young-Kyoon
    • 한국신재생에너지학회:학술대회논문집
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    • 2006.11a
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    • pp.529-532
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    • 2006
  • Recently the production of ethanol from lignocecllulosics has received much attention due to immense potential for conversion of renewable biometerials into biofuels and chemicals. Fomitopsis palustris causes a typycal brown-rot and is unusual in that it rapidly depolymerize the cellulose in wood without removing the surrounding lignin that normally prevents microbial attack. This study demonstrated that the brown rot basidiomycete F. palustris was able to degrade crystalline cellulose. This fungus could also produce the three major cellulases (BGL, EXG and EG) when the cells were grown on 2.0% Avicel. The fungus was able to degrade both the crystalline and amorphous forms of cellulose from woody biomasses. Moreover, we found that this fungus has the processive EG like CBH which are able to degrade the crystalline region of cellulose. To establish the cellulase system in relation with degradation of woody biomass, we performed that purification, characterization and molecular cloning of a BGL, EGs and GLA from F. palustris grown on Avicel.

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Biochemical Characterization of Human Foamy Virus Integrase (인간 포미바이러스 인테그라제의 생화학적 특성)

  • 강승이;오수아;이학성;한성태;서진욱;신차균
    • YAKHAK HOEJI
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    • v.48 no.1
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    • pp.13-19
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    • 2004
  • A bacterial expression vector for the human foamy virus (HFV) integrase was constructed and expressed in Escherichia coli. By two-step purification using a nickel-chelated column and a SP-sepharose chromatography; the HFV into-grase protein of 43 kDa was purified to near homogeneity, and used to investigate biochemical characteristics of the enzymatic activities, such as endonucleolytic and disintegration activities. Oligonucleotide substrates were specifically and efficiently cleaved by the purifed HFV integrase in the presence of Mn $^{+2}$, but not in the presence of Mg $^{+2}$, indicating that the HFV integrase is not able to use Mg $^{+2}$ as a cofactor Endonucleolytic reaction was almost completed in 60 min at 37 $^{\circ}C$. In addition, the maximum enzymatic activities were observed at 5 mM Mn $^{+2}$ in the buffer of which pH was from 7.0 to 9.0. The endonucleolytic activities were dose-dependently blocked in the addition of baicalein or chicolic acid which is a well-known inhibitor of human immunodeficiency virus integrase.

Fungal Distribution in Traditional Meju and Characterization of Isolated Strains

  • Ye-Eun Son;Ye-Jin Kang;Sun-Young Choi;Yoon-Kyung Choi;Ju-Eun Lee;Junyoung Kim;Hee-Soo Park
    • The Korean Journal of Mycology
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    • v.51 no.3
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    • pp.219-227
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    • 2023
  • This study was conducted to analyze the distribution and characteristics of fungal species in meju using the traditional method. Fungal distribution in meju was investigated using metagenomic and morphological analyses, based on which Aspergillus flavus/oryzae strains were identified as the dominant fungi in all meju samples, followed by Pichia, Rhizopus and Lichtheimia spp. As A. flavus/oryzae was dominant, we further evaluated the aflatoxin production ability and enzymatic activity of the isolates. Thin-layer chromatography and polymerase chain reaction revealed that the A. flavus/oryzae strains isolated from meju are non-aflatoxigenic fungi. Based on the analyses of amylase and protease activities, strains with high activities of amylase or protease were identified, which are proposed to be used as starters for meju fermentation.

New Insights in Arachidonate Cascade: Biochemical Characterization and Biological Significance of Three Distinct Prostaglandin E Synthases

  • Kudo, Ichiro
    • Proceedings of the PSK Conference
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    • 2003.04a
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    • pp.111-113
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    • 2003
  • Biosynthsis of prostaglandin E2 (PGE2), the most common prostanoid with potent and diverse bio-activities, is regulated by three sequential enzymatic steps composed of phospholipase A2, cyclooxygenase (COX), and prostaglandin E synthase (PGES). Recently, three distinct PGESs have been identified; two of them are membrane-bound enzymes, mPGES-1 and mPGES-2, and the third one is a cytosolic enzyme, cPGES. (omitted)

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A method of isolation and characterization of canine endometrial-derived mesenchymal stem cells

  • Mi Kyung Park;Kun Ho Song
    • Korean Journal of Veterinary Service
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    • v.46 no.2
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    • pp.157-160
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    • 2023
  • Endometrial tissue is a known source of mesenchymal stem cells (MSCs). We isolated canine endometrial stem cells from canine endometrial tissues using an enzymatic method and confirmed the immunophenotype of mesenchymal stem cells and multilineage differentiation. Canine endometrial tissues were obtained from canine ovariohysterectomy surgery and isolated using 0.2% collagenase type I. We measured the immunophenotype of stem cells using flow cytometry. To confirm the differentiation ability, a trilineage differentiation assay was conducted. In this study, canine endometrialderived MSCs (cEM-MSCs) were isolated by enzyme treatment and showed a spindle-shaped morphology under a microscope. Moreover, cEM-MSCs showed a trilineage differentiation ability. In this study, the canine endometrium was a good source of MSCs.

Determination of Antigenicity and Characterization of Proteinase from Tissue Invading Nematode Larvae (조직기생 선충류 유충에서 분리한 단백 분해 효소의 특성 및 항원성 검토)

  • Rim, Han-Jong;Joo, Kyeong-Hwan;Choi, Sung-A;Lee, Hye-Jeong;Joo, Chong-Yoon;Chung, Myung-Sook
    • Journal of agricultural medicine and community health
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    • v.22 no.1
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    • pp.61-74
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    • 1997
  • In case of tissue invading nematode, proteolytic enzyme was required at their parasitic life. Proteinases obtained from these parasites(Toxocara canis, Ansakis spp. and Trichinella spiralis) were extracted, isolated and further purified. And then the analysis for activity and inhibitory effect of proteinases were performed by appropriate substrate. Determination of protein as a circulating antigen was done in use of infected animal serum with above parasites, respectively. For above experimental objects, following procedures were performed. First, enzymatic activity was measured in use of azocasein and inhibitory effect of porteinase were studied by various inhibitors. Second, partially purified proteins containing enzymatic activity were obtained by ion exchange chromatography, ultrafiltration and electrophoretic elution. Third, role of the partially purified protein as a circulating antigen. The results obtained were as follows : 1. Enzymatic activity of each nematode proteinase was varied according to pH. Optimal pH of Toxocara canis, Ansakis spp. and Trichinella spiralis were pH 6.0, pH 5.5 and pH 6.5, respectively. The optimal molarity of buffer was 0.1M phosphate buffer. Although little difference between these proteinases was observed, temperature stability was at least maintained at $4^{\circ}C$ until 5 days. 2. In case of Ansakis spp. and Toxocara canis, enzymatic activity of these proteinases was considerably inhibited by Leupeptin and EDTA. For maximum enzymatic activity of above proteinases, it was required that cysteine residue of enzyme should be protected. And it was suggested that metallo type was contained in enzyme active site. Proteinase of Trichinella spiralis contained metallo type also. 3. Although partial purification was performed in Ansakis spp. and Toxocara canis, proteins maintaining enzymatic activity were identified as a circulating antigen. From SDS-PAGE and immunoblot, 25 kDa was presented in Ansakis spp.. Specific antigen of Toxocara cains was 110 kDa protein fraction. 55 and 42 kDa proteins were reacted with normal serum. Trichinella spiralis 60 kDa protein fraction was successfully purified from excretory materials in culture. As a result of immune-reaction with Trichinella spiralis infected serum, highly purified 60 kDa protein was maintained antigenicity until final purification step.

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Enzymatic Isolation of Husk from Barley Kernel (효소적 방법에 의한 보리피층의 분리)

  • Ahn, Byung-Yoon;Ahn, Kyung-Hay;Jo, Jae-Sun;Kwon, Tai-Wan;Cheigh, Hong-Sik
    • Korean Journal of Food Science and Technology
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    • v.16 no.2
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    • pp.254-256
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    • 1984
  • The amounts of husk materials from barley kernel were determined by an enzymatic method and compared with the values determined by conventional methods involving acid or alkaline treatments. The enzymatic method consists of boiling in distilled water and pressing to help squeeze out the gelatinized starch from the husk matrix, and enzymatic removal of starch by ${\alpha}-amylase$ and weighing the residual husk materials after washing 3 times with hot water and then drying at $95^{\circ}C$. Husk materials amounted about 15 of the covered barley (Gangbori and Olbori) and 10-12% of naked variety (Backdong and Sedohadaga) and the values were always somewhat higher than those obtained by the conventional methods. The husk materials prepared by the enzymatic procedure contained protein 4-8%, lipid 5-10%, ash 0.2-0.6% and crude fiber 20-40%. Although it took longer time, the enzymatic procedures can provide more intack husk materials for further characterization of the materials.

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Characterization of Undaria pinnatifida Root Enzymatic Extracts Using Crude Enzyme from Shewanella oneidensis PKA 1008 and Its Anti-Inflammatory Effect

  • Xu, Xiaotong;Jeong, So-Mi;Lee, Ji-Eun;Kang, Woo-Sin;Ryu, Si-Hyeong;Kim, Kwangwook;Byun, Eui-Hong;Cho, Young-Je;Ahn, Dong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.30 no.1
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    • pp.79-84
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    • 2020
  • This study investigated the characterization and functionality of Undaria pinnatifida root (UPT) extracts, degraded using a crude enzyme from Shewanella oneidensis PKA1008. To obtain the optimum degrading conditions, the UPT was mixed with alginate degrading enzymes from S. oneidensis PKA 1008 and was incubated at 30℃ for 0, 3, 6, 12, 24, and 48 h. The alginate degrading ability of these enzymes was then evaluated by measuring the reducing sugar, viscosity, pH and chromaticity. Enzymatic extract at 24 h revealed the highest alginate degrading ability and the lowest pH value. As the incubation time increased, the lightness (L ) also decreased and was measured at its lowest value, 39.84, at 12 hours. The redness and yellowness increased gradually to 10.27 at 6 h and to 63.95 at 3 h, respectively. Moreover, the alginate oligosaccharides exhibited significant anti-inflammatory activity. These results indicate that a crude enzyme from S. oneidensis PKA 1008 can be used to enhance the polysaccharide degradation of UPT and the alginate oligosaccharides may also enhance the anti-inflammatory effect.