• Title/Summary/Keyword: partial identification

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Isolation and Enzyme Production of a Mannanase-producing Strain, Bacillus sp. WL-3. (Mannanase를 생산하는 Bacillus sp. WL-3 균주의 분리와 효소 생산성)

  • 오영필;이정민;조기행;윤기홍
    • Microbiology and Biotechnology Letters
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    • v.30 no.3
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    • pp.247-252
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    • 2002
  • A bacterium producing the extracellular mannanase was isolated from Korean formented food and has been identified as a member of the genus Bacillus from the result of the phylogenic analysis based on partial 165 rRNA sequences. The isolate, named Bacillus sp. WL-3, was shown to be similar to B. subtilis strain on the basis of its biochemical properties. The mannanase of culture supematant was the most active at $55^{\circ}C$ and pH 6.0. The additional carbohydrates including u-cellulose, avicel, oat spelt xylan, guar gum and locust bean gum (LBG) increased the mannanase productivity. Especially, the maximum mannanase productivity was reached 65.5 U/ml in LB medium supplemented with 0.5% (w/v) LBG, which was 131-folds more than that in LB medium. It was sug-gested that the increase of mannanase production was owing to induction of mannanase biosynthesis by LBG hydrolysates transported following initial hydrolysis by extracellular mannanase during the cell growth. The molec-ular weight of WL-3 mannanase was estimated to approximately 38.0 kDa by zymogram on SDS-PAGE.

Purification and Characterization of an Extracellular ${\beta}$-Glucosidase Produced by Phoma sp. KCTC11825BP Isolated from Rotten Mandarin Peel

  • Choi, Jung-Youn;Park, Ah-Reum;Kim, Yong-Jin;Kim, Jae-Jin;Cha, Chang-Jun;Yoon, Jeong-Jun
    • Journal of Microbiology and Biotechnology
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    • v.21 no.5
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    • pp.503-508
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    • 2011
  • A ${\beta}$-glucosidase from Phoma sp. KCTC11825BP isolated from rotten mandarin peel was purified 8.5-fold with a specific activity of 84.5 U/mg protein. The purified enzyme had a molecular mass of 440 kDa with a subunit of 110 kDa. The partial amino acid sequence of the purified ${\beta}$-glucosidase evidenced high homology with the fungal ${\beta}$- glucosidases belonging to glycosyl hydrolase family 3. Its optimal activity was detected at pH 4.5 and $60^{\circ}C$, and the enzyme had a half-life of 53 h at $60^{\circ}C$. The $K_m$ values for p-nitrophenyl-${\beta}$-D-glucopyranoside and cellobiose were 0.3 mM and 3.2 mM, respectively. The enzyme was competitively inhibited by both glucose ($K_i$=1.7 mM) and glucono-${\delta}$-lactone ($K_i$=0.1 mM) when pNPG was used as the substrate. Its activity was inhibited by 41% by 10 mM $Cu^{2+}$ and stimulated by 20% by 10 mM $Mg^{2+}$.

Analysis of oligosaccharides from Panax ginseng by using solid-phase permethylation method combined with ultra-high-performance liquid chromatography-Q-Orbitrap/mass spectrometry

  • Li, Lele;Ma, Li;Guo, Yunlong;Liu, Wenlong;Wang, Yang;Liu, Shuying
    • Journal of Ginseng Research
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    • v.44 no.6
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    • pp.775-783
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    • 2020
  • Background: The reports about valuable oligosaccharides in ginseng are quite limited. There is an urgent need to develop a practical procedure to detect and analyze ginseng oligosaccharides. Methods: The oligosaccharide extracts from ginseng were permethylated by solid-phase methylation method and then were analyzed by ultra-high-performance liquid chromatography-Q-Orbitrap/MS. The sequence, linkage, and configuration information of oligosaccharides were determined by using accurate m/z value and tandem mass information. Several standard references were used to further confirm the identification. The oligosaccharide composition in white ginseng and red ginseng was compared using a multivariate statistical analysis method. Results: The nonreducing oligosaccharide erlose among 12 oligosaccharides identified was reported for the first time in ginseng. In the comparison of the oligosaccharide extracts from white ginseng and red ginseng, a clear separation was observed in the partial least squares-discriminate analysis score plot, indicating the sugar differences in these two kinds of ginseng samples. The glycans with variable importance in the projection value large than 1.0 were considered to contribute most to the classification. The contents of oligosaccharides in red ginseng were lower than those in white ginseng, and the contents of maltose, maltotriose, maltotetraose, maltopentaose, maltohexaose, maltoheptaose, maltooctaose, maltononaose, sucrose, and erlose decreased significantly (p < 0.05) in red ginseng. Conclusion: A solid-phase methylation method combined with liquid chromatography-tandem mass spectrometry was successfully applied to analyze the oligosaccharides in ginseng extracts, which provides the possibility for holistic evaluation of ginseng oligosaccharides. The comparison of oligosaccharide composition of white ginseng and red ginseng could help understand the differences in pharmacological activities between these two kinds of ginseng samples from the perspective of glycans.

Proteomics Analysis of Gastric Epithelial AGS Cells Infected with Epstein-Barr Virus

  • Ding, Yong;Li, Xiao-Rong;Yang, Kai-Yan;Huang, Li-Hua;Hu, Gui;Gao, Kai
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.1
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    • pp.367-372
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    • 2013
  • Effects of the Epstein-Barr virus (EBV) on cellular protein expression are essential for viral pathogenesis. To characterize the cellular response to EBV infection, differential proteomes of gastric epithelial AGS cells were analyzed with two-dimensional gel electrophoresis (2-DE) followed by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) and liquid chromatography electrospray/ionization ion trap (LC-ESI-IT) mass spectrometry identification. Mass spectrometry identified 9 altered cellular proteins, including 5 up-regulated and 4 down-regulated proteins after EBV infection. Notably 2-DE analysis revealed that EBV infection induced increased expression of heat shock cognate 71 kDa protein, actin cytoplasmic 1, pyridoxine-5'-phosphate oxidase, caspase 9, and t-complex protein 1 subunit alpha. In addition, EBV infection considerably suppressed those cellular proteins of zinc finger protein 2, cyclin-dependent kinase 2, macrophage-capping protein, and growth/differentiation factor 11. Furthermore, the differential expressional levels of partial proteins (cyclin-dependent kinase 2 and caspase 9) were confirmed by Western blot analysis.Thus, this work effectively provided useful protein-related information to facilitate further investigation of the mechanisms underlying EBV infection and pathogenesis.

Identification of Design Attributes of the Affective Expressions for Movie Making (영화의 감성만족도 측정을 위한 시.청각적 영향 요인의 체계적 도출)

  • Kim, In-Ki;Kim, Ji-Ho;Chang, Woo-Jin;Lee, Cheol;Yun, Myung-Hwan
    • 한국HCI학회:학술대회논문집
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    • 2007.02a
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    • pp.143-149
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    • 2007
  • 영상은 동적인 시각 이미지와 청각의 결합에 의해 감성적인 반응을 유도한다. 다양한 영상 기법을 통하여 감성적 반응의 극대화를 추구하는 영화는 영상의 시청각적 요소들을 감성의 관점에서 효과적으로 설계하는데 본보기가 된다. 그러나, 제품의 설계속성들에 대한 감성적 평가결과를 모형화하는 감성공학적 관점에서 볼 때 영화는 시청각적 자극의 수준이 극히 다양하고 동적인 경험재로 모형화의 어려움이 있다. 본 연구에서는 영화의 감성 모형을 구축하기 위한 사전연구의 단계로 영화에서의 시청각적 요인들을 문헌조사를 통해 수집, 정리, 선별하고 이러한 시청각적 요인들 중에 영화를 관람하는 관객의 감성적, 인지적 반응에 영향을 주는 유효한 요인들을 객관적이고 체계적으로 탐색하고자 하였다. 이를 위해, 감성 및 인지적 반응의 변화를 생체신호를 통해 측정하는 한편, 생체신호의 측정 시 사용된 영화의 시청각적 자극요인을 Video/Audio Processing방법에 의해 연속적인 수치로 정량화하였다. 생체신호와 정량화된 시청각적 자극요인을 동기화하고 통계적으로 분석함으로써, 생체신호의 반응과 시청각적 자극요인과의 인과관계를 통계적으로 신뢰성있는 수준에서 검증하고자 하였다. 생체신호를 종속변수로, 시청각적 자극요인을 독립변수로 하는 896개의 부분선형회귀모형(Partial Linear Regression Model)들 중 통계적으로 유의한 선형관계에 있는 경우의 빈도분석에 의하면, 시각적 요인들 중에는 밝기(Brightness), 대비(Contrast), 색상(Color), 움직임(Motion), 장면전환속도(Shot change Rate), 주요대상의 상대적 크기가, 청각적 요인들 중에는 Peak주파수, Peak주파수의 음량, 평균음량, 소음비(Sound-to-Noise Ratio)가 생체신호의 변화에 통계적으로 유의한 영향을 주는 것으로 나타났다. 이는, 위의 시청각적 자극 요인들은 특히 관객의 감성 및 인지적인 반응에 유의한 영향을 주는 요소로 작용할 수 있음을 시사하고 있다. 이를 토대로, 위의 시청각적 자극 요인들이 가지는 다양한 조합들을 설명변수로 하는 통계적인 영화의 감성 모형을 구축할 수 있을 것으로 기대한다.

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Screening and Identification of Monascus Strain Producing Monacolin K (Monacolin K 대량생산 Monascus 균주의 탐색 및 동정)

  • 곽은정;이효민;임성일
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.1
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    • pp.164-169
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    • 2004
  • We had screened the Monascus strain capable of producing monacolin K dominantly among 29 Monascus strains. Red yeast rice was prepared by culturing each Monascus sp. with 200 g of steamed rice (12$0^{\circ}C$, 20 min) at 3$0^{\circ}C$ for 10 days and drying at 8$0^{\circ}C$ for 20 min. As a result, red yeast rice cultured by M. purpureus ATCC16457, M. purpureu IFO 32316, M. purpureus IFO 32228, M. kaoliang ATCC 46595 and M. kaoliang ATCC 46596 produced lots of red pigment and monacolin K. An unidentified Monascus sp. showed the highest productivityof red pigment and monacolin K among 29 Monascus strains. Its production of red pigment and monacolin K was 1.3∼39 times and 2.4∼8 times higher than other strains, respectively. Although the morphological characteristics of unidentified Monascus strain were a little different from the typical M. purpureus, it was identified as M. purpureus CBS 281.34 from the result of sequencing of ITS (Internal transcribed spacer) and 28S ribosomal RNA (partial).

cDNA Microarray Analysis of the Differential Gene Expression in the Neuropathic Pain and Electroacupuncture Treatment Models

  • Ko, Je-Sang;Na, Doe-Sun;Lee, Young-Han;Shin, Soon-Young;Kim, Ji-Hoon;Hwang, Byung-Gil;Min, Byung-Il;Park, Dong-Suk
    • BMB Reports
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    • v.35 no.4
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    • pp.420-427
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    • 2002
  • Partial nerve injury is the main cause of neuropathic pain disorders in humans. Acupuncture has long been used to relieve pain. It is known to relieve pain by controlling the activities of the autonomic nervous system. Although the mechanism of neuropathic pain and analgesic effects of electroacupuncture (EA) have been studied in a rat model system, its detailed mechanism at the molecular level remains unclear. To identify genes that might serve as either markers or explain these distinct biological functions, a cDNA microarray analysis was used to compare the expression of 8,400 genes among three sample groups. Messenger RNAs that were pooled from the spinal nerves of 7 normal. 7 neuropathic pain, and 7 EA treatment rat models were compared. Sixty-eight genes were differentially expressed more than 2-fold in the neuropathic rat model when compared to the normal, and restored to the normal expression level after the EA treatment. These genes are involved in a number of biological processes, including the signal transduction, gene expression, and nociceptive pathways. Confirmation of the differential gene expression was performed by a dot-blot analysis. Dot-blotting results showed that the opioid receptor sigma was among those genes. This indicates that opioid-signaling events are involved in neuropathic pain and the analgesic effects of EA. The potential application of these data include the identification and characterization of signaling pathways that are involved in the EA treatment, studies on the role of the opioid receptor in neuropathic pain, and further exploration on the role of selected identified genes in animal models.

Molecular Genetic Findings of Spirometra decipiens and S. ranarum in Korea

  • Jeon, Hyeong-Kyu;Huh, Sun;Sohn, Woon-Mok;Chai, Jong-Yil;Eom, Keeseon S.
    • Parasites, Hosts and Diseases
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    • v.56 no.4
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    • pp.359-364
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    • 2018
  • The taxonomy of Spirometra species has been controversial despite the medical and veterinary importance. Currently, only a few Spirometra species are considered valid species in the genus Spirometra. In the present study, the distribution of Spirometra species obtained from animals in Korea were identified by molecular analysis of the mitochondrial cytochrome c oxidase I (cox1) gene. A total of 28 Spirometra species specimens were analyzed. These were all collected between 1973 and 2008 in the Republic of Korea. Mitochondrial cox1 sequences were examined for a total of 28 specimens comprising 14 S. decipiens and 14 S. ranarum. The difference in partial cox1 sequences (316 bp) between S. erinaceieuropaei (KJ599680) and S. ranarum (this study) was 9.3%, while that between S. decipiens (KJ599679) and S. ranarum (this study) was 2.2%. Genetic analyses identified 2 Spirometra species in animals such as cat, leopard cat, dog, duck and snake in Korea as S. decipiens and S. ranarum. S. decipiens and S. ranarum were present in Gyeongnam Province (P), Jeonnam P, Gangwon P, Chungbuk P, and Seoul. S. decipiens was found in tadpoles, snakes, ducks, cats, leopard cats and dogs, while S. ranarum was found in cats and dogs. The ratio of S. decipiens:S. ranarum calculated from the molecular data was 14:14 (or 1:1). These results indicate that S. decipiens and S. ranarum are sympatrically distributed in Korea.

Isolation and Identification of Endophytic Bacteria with Plant Growth Promoting Activity and Biocontrol Potential from Wild Pistachio Trees

  • Etminani, Faegheh;Harighi, Behrouz
    • The Plant Pathology Journal
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    • v.34 no.3
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    • pp.208-217
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    • 2018
  • In this study, samples were collected from the leaves and stems of healthy wild Pistachio trees (Pistacia atlantica L.) from various locations of Baneh and Marivan regions, Iran. In total, 61 endophytic bacteria were isolated and grouped according to phenotypic properties. Ten selected isolates from each group were further identified by partial sequencing of the 16S rRNA gene. Based on the results, isolates were identified as bacteria belonging to Pseudomonas, Stenotrophomonas, Bacillus, Pantoea and Serratia genus. The ability of these isolates was evaluated to phytohormone production such as auxin and gibberellin, siderophore production, phosphate solubilization, atmospheric nitrogen fixation, protease and hydrogen cyanide production. All strains were able to produce the plant growth hormone auxin and gibberellin in different amounts. The majority of strains were able to solubilize phosphate. The results of atmospheric nitrogen fixation ability, protease and siderophore production were varied among strains. Only Ba66 could produce a low amount of hydrogen cyanide. The results of biocontrol assay showed that Pb78 and Sp15 strains had the highest and lowest inhibition effects on bacterial plant pathogens, Pseudomonas syringae pv. syringae Pss20 and Pseudomonas tolaasii Pt18 under in vitro condition. Pb3, Pb24 and Pb71 strains significantly promote root formation on carrot slices. To our knowledge this is the first report of the isolation of endophytic bacterial strains belonging to Pantoea, Bacillus, Pseudomonas, Serratia and Stenotrophomonas genus from wild pistachio trees with plant growth promoting potential and biocontrol activity.

A CONSTRUCTION METHOD OF MULTIPLE CONTROL SYSTEMS USING PARTIAL KNOWLEDGE UPON SYSTEM DYNAMICS

  • Yoshisara, Ikuo;Indaba, Masaaki;Aoyama, Tomoo;Yasunaga, Moritoshi
    • 제어로봇시스템학회:학술대회논문집
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    • 1999.10a
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    • pp.73-78
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    • 1999
  • This paper presents an effective construction method of adaptive multiple control systems utilizing some knowledge upon the plants. The adaptive multiple control system operates plants un-der widely changing environmental conditions. The adaptive multiple control system is composed of a family of candidate controllers together with a supervisor. The system does not require any identification schemes of environmental conditions. Monitoring outputs of the plant, the supervisor switches from one candidate controller to another, The basic ideas of adaptation are as follows: (1)each candidate controller is prepared for each environmental condition in advance; (2)the supervise. applies a sequence of speculative controls to the plant with candidate controllers just after the start of control or just after the detection of a change in the environmental condition. Each candidate controller can keep the system stable during one-step period of the speculative control and the most appropriate candidate controller for the environmental condition to which the system is exposed can be selected before the last trial of speculative control step comes to an end. We proposed a construction method of adaptive multiple control system without any knowledge of plant dynamics and applied the method to a cart-pole balancing problem and a vehicle anti skid braking system. In real applications, as we can often easily obtain a piece of knowledge upon plant dynamics beforehand, we intend to extend the method such that multiple control systems can be efficiently designed using the knowledge. We apply the new idea to the cart-pole balancing problem with variable length of the pole. The simulation experiments lead us to the conclusion that the new attempt can reduce the manpower to design the candidate controllers for adaptive multiple control systems.

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