• Title/Summary/Keyword: Modification

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Sesquiterpenoids Bioconversion Analysis by Wood Rot Fungi

  • Lee, Su-Yeon;Ryu, Sun-Hwa;Choi, In-Gyu;Kim, Myungkil
    • 한국균학회소식:학술대회논문집
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    • 2016.05a
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    • pp.19-20
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    • 2016
  • Sesquiterpenoids are defined as $C_{15}$ compounds derived from farnesyl pyrophosphate (FPP), and their complex structures are found in the tissue of many diverse plants (Degenhardt et al. 2009). FPP's long chain length and additional double bond enables its conversion to a huge range of mono-, di-, and tri-cyclic structures. A number of cyclic sesquiterpenes with alcohol, aldehyde, and ketone derivatives have key biological and medicinal properties (Fraga 1999). Fungi, such as the wood-rotting Polyporus brumalis, are excellent sources of pharmaceutically interesting natural products such as sesquiterpenoids. In this study, we investigated the biosynthesis of P. brumalis sesquiterpenoids on modified medium. Fungal suspensions of 11 white rot species were inoculated in modified medium containing $C_6H_{12}O_6$, $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ for 20 days. Cultivation was stopped by solvent extraction via separation of the mycelium. The metabolites were identified as follows: propionic acid (1), mevalonic acid lactone (2), ${\beta}$-eudesmane (3), and ${\beta}$-eudesmol (4), respectively (Figure 1). The main peaks of ${\beta}$-eudesmane and ${\beta}$-eudesmol, which were indicative of sesquiterpene structures, were consistently detected for 5, 7, 12, and 15 days These results demonstrated the existence of terpene metabolism in the mycelium of P. brumalis. Polyporus spp. are known to generate flavor components such as methyl 2,4-dihydroxy-3,6-dimethyl benzoate; 2-hydroxy-4-methoxy-6-methyl benzoic acid; 3-hydroxy-5-methyl phenol; and 3-methoxy-2,5-dimethyl phenol in submerged cultures (Hoffmann and Esser 1978). Drimanes of sesquiterpenes were reported as metabolites from P. arcularius and shown to exhibit antimicrobial activity against Gram-positive bacteria such as Staphylococcus aureus (Fleck et al. 1996). The main metabolites of P. brumalis, ${\beta}$-Eudesmol and ${\beta}$-eudesmane, were categorized as eudesmane-type sesquiterpene structures. The eudesmane skeleton could be biosynthesized from FPP-derived IPP, and approximately 1,000 structures have been identified in plants as essential oils. The biosynthesis of eudesmol from P. brumalis may thus be an important tool for the production of useful natural compounds as presumed from its identified potent bioactivity in plants. Essential oils comprising eudesmane-type sesquiterpenoids have been previously and extensively researched (Wu et al. 2006). ${\beta}$-Eudesmol is a well-known and important eudesmane alcohol with an anticholinergic effect in the vascular endothelium (Tsuneki et al. 2005). Additionally, recent studies demonstrated that ${\beta}$-eudesmol acts as a channel blocker for nicotinic acetylcholine receptors at the neuromuscular junction, and it can inhibit angiogenesis in vitro and in vivo by blocking the mitogen-activated protein kinase (MAPK) signaling pathway (Seo et al. 2011). Variation of nutrients was conducted to determine an optimum condition for the biosynthesis of sesquiterpenes by P. brumalis. Genes encoding terpene synthases, which are crucial to the terpene synthesis pathway, generally respond to environmental factors such as pH, temperature, and available nutrients (Hoffmeister and Keller 2007, Yu and Keller 2005). Calvo et al. described the effect of major nutrients, carbon and nitrogen, on the synthesis of secondary metabolites (Calvo et al. 2002). P. brumalis did not prefer to synthesize sesquiterpenes under all growth conditions. Results of differences in metabolites observed in P. brumalis grown in PDB and modified medium highlighted the potential effect inorganic sources such as $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ on sesquiterpene synthesis. ${\beta}$-eudesmol was apparent during cultivation except for when P. brumalis was grown on $MgSO_4$-free medium. These results demonstrated that $MgSO_4$ can specifically control the biosynthesis of ${\beta}$-eudesmol. Magnesium has been reported as a cofactor that binds to sesquiterpene synthase (Agger et al. 2008). Specifically, the $Mg^{2+}$ ions bind to two conserved metal-binding motifs. These metal ions complex to the substrate pyrophosphate, thereby promoting the ionization of the leaving groups of FPP and resulting in the generation of a highly reactive allylic cation. Effect of magnesium source on the sesquiterpene biosynthesis was also identified via analysis of the concentration of total carbohydrates. Our current study offered further insight that fungal sesquiterpene biosynthesis can be controlled by nutrients. To profile the metabolites of P. brumalis, the cultures were extracted based on the growth curve. Despite metabolites produced during mycelia growth, there was difficulty in detecting significant changes in metabolite production, especially those at low concentrations. These compounds may be of interest in understanding their synthetic mechanisms in P. brumalis. The synthesis of terpene compounds began during the growth phase at day 9. Sesquiterpene synthesis occurred after growth was complete. At day 9, drimenol, farnesol, and mevalonic lactone (or mevalonic acid lactone) were identified. Mevalonic acid lactone is the precursor of the mevalonic pathway, and particularly, it is a precursor for a number of biologically important lipids, including cholesterol hormones (Buckley et al. 2002). Farnesol is the precursor of sesquiterpenoids. Drimenol compounds, bi-cyclic-sesquiterpene alcohols, can be synthesized from trans-trans farnesol via cyclization and rearrangement (Polovinka et al. 1994). They have also been identified in the basidiomycota Lentinus lepideus as secondary metabolites. After 12 days in the growth phase, ${\beta}$-elemene caryophyllene, ${\delta}$-cadiene, and eudesmane were detected with ${\beta}$-eudesmol. The data showed the synthesis of sesquiterpene hydrocarbons with bi-cyclic structures. These compounds can be synthesized from FPP by cyclization. Cyclic terpenoids are synthesized through the formation of a carbon skeleton from linear precursors by terpene cyclase, which is followed by chemical modification by oxidation, reduction, methylation, etc. Sesquiterpene cyclase is a key branch-point enzyme that catalyzes the complex intermolecular cyclization of the linear prenyl diphosphate into cyclic hydrocarbons (Toyomasu et al. 2007). After 20 days in stationary phase, the oxygenated structures eudesmol, elemol, and caryophyllene oxide were detected. Thus, after growth, sesquiterpenes were identified. Per these results, we showed that terpene metabolism in wood-rotting fungi occurs in the stationary phase. We also showed that such metabolism can be controlled by magnesium supplementation in the growth medium. In conclusion, we identified P. brumalis as a wood-rotting fungus that can produce sesquiterpenes. To mechanistically understand eudesmane-type sesquiterpene biosynthesis in P. brumalis, further research into the genes regulating the dynamics of such biosynthesis is warranted.

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Perceptional Change of a New Product, DMB Phone

  • Kim, Ju-Young;Ko, Deok-Im
    • Journal of Global Scholars of Marketing Science
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    • v.18 no.3
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    • pp.59-88
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    • 2008
  • Digital Convergence means integration between industry, technology, and contents, and in marketing, it usually comes with creation of new types of product and service under the base of digital technology as digitalization progress in electro-communication industries including telecommunication, home appliance, and computer industries. One can see digital convergence not only in instruments such as PC, AV appliances, cellular phone, but also in contents, network, service that are required in production, modification, distribution, re-production of information. Convergence in contents started around 1990. Convergence in network and service begins as broadcasting and telecommunication integrates and DMB(digital multimedia broadcasting), born in May, 2005 is the symbolic icon in this trend. There are some positive and negative expectations about DMB. The reason why two opposite expectations exist is that DMB does not come out from customer's need but from technology development. Therefore, customers might have hard time to interpret the real meaning of DMB. Time is quite critical to a high tech product, like DMB because another product with same function from different technology can replace the existing product within short period of time. If DMB does not positioning well to customer's mind quickly, another products like Wibro, IPTV, or HSPDA could replace it before it even spreads out. Therefore, positioning strategy is critical for success of DMB product. To make correct positioning strategy, one needs to understand how consumer interprets DMB and how consumer's interpretation can be changed via communication strategy. In this study, we try to investigate how consumer perceives a new product, like DMB and how AD strategy change consumer's perception. More specifically, the paper segment consumers into sub-groups based on their DMB perceptions and compare their characteristics in order to understand how they perceive DMB. And, expose them different printed ADs that have messages guiding consumer think DMB in specific ways, either cellular phone or personal TV. Research Question 1: Segment consumers according to perceptions about DMB and compare characteristics of segmentations. Research Question 2: Compare perceptions about DMB after AD that induces categorization of DMB in direction for each segment. If one understand and predict a direction in which consumer perceive a new product, firm can select target customers easily. We segment consumers according to their perception and analyze characteristics in order to find some variables that can influence perceptions, like prior experience, usage, or habit. And then, marketing people can use this variables to identify target customers and predict their perceptions. If one knows how customer's perception is changed via AD message, communication strategy could be constructed properly. Specially, information from segmented customers helps to develop efficient AD strategy for segment who has prior perception. Research framework consists of two measurements and one treatment, O1 X O2. First observation is for collecting information about consumer's perception and their characteristics. Based on first observation, the paper segment consumers into two groups, one group perceives DMB similar to Cellular phone and the other group perceives DMB similar to TV. And compare characteristics of two segments in order to find reason why they perceive DMB differently. Next, we expose two kinds of AD to subjects. One AD describes DMB as Cellular phone and the other Ad describes DMB as personal TV. When two ADs are exposed to subjects, consumers don't know their prior perception of DMB, in other words, which subject belongs 'similar-to-Cellular phone' segment or 'similar-to-TV' segment? However, we analyze the AD's effect differently for each segment. In research design, final observation is for investigating AD effect. Perception before AD is compared with perception after AD. Comparisons are made for each segment and for each AD. For the segment who perceives DMB similar to TV, AD that describes DMB as cellular phone could change the prior perception. And AD that describes DMB as personal TV, could enforce the prior perception. For data collection, subjects are selected from undergraduate students because they have basic knowledge about most digital equipments and have open attitude about a new product and media. Total number of subjects is 240. In order to measure perception about DMB, we use indirect measurement, comparison with other similar digital products. To select similar digital products, we pre-survey students and then finally select PDA, Car-TV, Cellular Phone, MP3 player, TV, and PSP. Quasi experiment is done at several classes under instructor's allowance. After brief introduction, prior knowledge, awareness, and usage about DMB as well as other digital instruments is asked and their similarities and perceived characteristics are measured. And then, two kinds of manipulated color-printed AD are distributed and similarities and perceived characteristics for DMB are re-measured. Finally purchase intension, AD attitude, manipulation check, and demographic variables are asked. Subjects are given small gift for participation. Stimuli are color-printed advertising. Their actual size is A4 and made after several pre-test from AD professionals and students. As results, consumers are segmented into two subgroups based on their perceptions of DMB. Similarity measure between DMB and cellular phone and similarity measure between DMB and TV are used to classify consumers. If subject whose first measure is less than the second measure, she is classified into segment A and segment A is characterized as they perceive DMB like TV. Otherwise, they are classified as segment B, who perceives DMB like cellular phone. Discriminant analysis on these groups with their characteristics of usage and attitude shows that Segment A knows much about DMB and uses a lot of digital instrument. Segment B, who thinks DMB as cellular phone doesn't know well about DMB and not familiar with other digital instruments. So, consumers with higher knowledge perceive DMB similar to TV because launching DMB advertising lead consumer think DMB as TV. Consumers with less interest on digital products don't know well about DMB AD and then think DMB as cellular phone. In order to investigate perceptions of DMB as well as other digital instruments, we apply Proxscal analysis, Multidimensional Scaling technique at SPSS statistical package. At first step, subjects are presented 21 pairs of 7 digital instruments and evaluate similarity judgments on 7 point scale. And for each segment, their similarity judgments are averaged and similarity matrix is made. Secondly, Proxscal analysis of segment A and B are done. At third stage, get similarity judgment between DMB and other digital instruments after AD exposure. Lastly, similarity judgments of group A-1, A-2, B-1, and B-2 are named as 'after DMB' and put them into matrix made at the first stage. Then apply Proxscal analysis on these matrixes and check the positional difference of DMB and after DMB. The results show that map of segment A, who perceives DMB similar as TV, shows that DMB position closer to TV than to Cellular phone as expected. Map of segment B, who perceive DMB similar as cellular phone shows that DMB position closer to Cellular phone than to TV as expected. Stress value and R-square is acceptable. And, change results after stimuli, manipulated Advertising show that AD makes DMB perception bent toward Cellular phone when Cellular phone-like AD is exposed, and that DMB positioning move towards Car-TV which is more personalized one when TV-like AD is exposed. It is true for both segment, A and B, consistently. Furthermore, the paper apply correspondence analysis to the same data and find almost the same results. The paper answers two main research questions. The first one is that perception about a new product is made mainly from prior experience. And the second one is that AD is effective in changing and enforcing perception. In addition to above, we extend perception change to purchase intention. Purchase intention is high when AD enforces original perception. AD that shows DMB like TV makes worst intention. This paper has limitations and issues to be pursed in near future. Methodologically, current methodology can't provide statistical test on the perceptual change, since classical MDS models, like Proxscal and correspondence analysis are not probability models. So, a new probability MDS model for testing hypothesis about configuration needs to be developed. Next, advertising message needs to be developed more rigorously from theoretical and managerial perspective. Also experimental procedure could be improved for more realistic data collection. For example, web-based experiment and real product stimuli and multimedia presentation could be employed. Or, one can display products together in simulated shop. In addition, demand and social desirability threats of internal validity could influence on the results. In order to handle the threats, results of the model-intended advertising and other "pseudo" advertising could be compared. Furthermore, one can try various level of innovativeness in order to check whether it make any different results (cf. Moon 2006). In addition, if one can create hypothetical product that is really innovative and new for research, it helps to make a vacant impression status and then to study how to form impression in more rigorous way.

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Stratigraphic response to tectonic evolution of sedimentary basins in the Yellow Sea and adjacent areas (황해 및 인접 지역 퇴적분지들의 구조적 진화에 따른 층서)

  • Ryo In Chang;Kim Boo Yang;Kwak won Jun;Kim Gi Hyoun;Park Se Jin
    • The Korean Journal of Petroleum Geology
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    • v.8 no.1_2 s.9
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    • pp.1-43
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    • 2000
  • A comparison study for understanding a stratigraphic response to tectonic evolution of sedimentary basins in the Yellow Sea and adjacent areas was carried out by using an integrated stratigraphic technology. As an interim result, we propose a stratigraphic framework that allows temporal and spatial correlation of the sedimentary successions in the basins. This stratigraphic framework will use as a new stratigraphic paradigm for hydrocarbon exploration in the Yellow Sea and adjacent areas. Integrated stratigraphic analysis in conjunction with sequence-keyed biostratigraphy allows us to define nine stratigraphic units in the basins: Cambro-Ordovician, Carboniferous-Triassic, early to middle Jurassic, late Jurassic-early Cretaceous, late Cretaceous, Paleocene-Eocene, Oligocene, early Miocene, and middle Miocene-Pliocene. They are tectono-stratigraphic units that provide time-sliced information on basin-forming tectonics, sedimentation, and basin-modifying tectonics of sedimentary basins in the Yellow Sea and adjacent area. In the Paleozoic, the South Yellow Sea basin was initiated as a marginal sag basin in the northern margin of the South China Block. Siliciclastic and carbonate sediments were deposited in the basin, showing cyclic fashions due to relative sea-level fluctuations. During the Devonian, however, the basin was once uplifted and deformed due to the Caledonian Orogeny, which resulted in an unconformity between the Cambro-Ordovician and the Carboniferous-Triassic units. The second orogenic event, Indosinian Orogeny, occurred in the late Permian-late Triassic, when the North China block began to collide with the South China block. Collision of the North and South China blocks produced the Qinling-Dabie-Sulu-Imjin foldbelts and led to the uplift and deformation of the Paleozoic strata. Subsequent rapid subsidence of the foreland parallel to the foldbelts formed the Bohai and the West Korean Bay basins where infilled with the early to middle Jurassic molasse sediments. Also Piggyback basins locally developed along the thrust. The later intensive Yanshanian (first) Orogeny modified these foreland and Piggyback basins in the late Jurassic. The South Yellow Sea basin, however, was likely to be a continental interior sag basin during the early to middle Jurassic. The early to middle Jurassic unit in the South Yellow Sea basin is characterized by fluvial to lacustrine sandstone and shale with a thick basal quartz conglomerate that contains well-sorted and well-rounded gravels. Meanwhile, the Tan-Lu fault system underwent a sinistrai strike-slip wrench movement in the late Triassic and continued into the Jurassic and Cretaceous until the early Tertiary. In the late Jurassic, development of second- or third-order wrench faults along the Tan-Lu fault system probably initiated a series of small-scale strike-slip extensional basins. Continued sinistral movement of the Tan-Lu fault until the late Eocene caused a megashear in the South Yellow Sea basin, forming a large-scale pull-apart basin. However, the Bohai basin was uplifted and severely modified during this period. h pronounced Yanshanian Orogeny (second and third) was marked by the unconformity between the early Cretaceous and late Eocene in the Bohai basin. In the late Eocene, the Indian Plate began to collide with the Eurasian Plate, forming a megasuture zone. This orogenic event, namely the Himalayan Orogeny, was probably responsible for the change of motion of the Tan-Lu fault system from left-lateral to right-lateral. The right-lateral strike-slip movement of the Tan-Lu fault caused the tectonic inversion of the South Yellow Sea basin and the pull-apart opening of the Bohai basin. Thus, the Oligocene was the main period of sedimentation in the Bohai basin as well as severe tectonic modification of the South Yellow Sea basin. After the Oligocene, the Yellow Sea and Bohai basins have maintained thermal subsidence up to the present with short periods of marine transgressions extending into the land part of the present basins.

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