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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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The Impact of Market Environments on Optimal Channel Strategy Involving an Internet Channel: A Game Theoretic Approach (시장 환경이 인터넷 경로를 포함한 다중 경로 관리에 미치는 영향에 관한 연구: 게임 이론적 접근방법)

  • Yoo, Weon-Sang
    • Journal of Distribution Research
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    • v.16 no.2
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    • pp.119-138
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    • 2011
  • Internet commerce has been growing at a rapid pace for the last decade. Many firms try to reach wider consumer markets by adding the Internet channel to the existing traditional channels. Despite the various benefits of the Internet channel, a significant number of firms failed in managing the new type of channel. Previous studies could not cleary explain these conflicting results associated with the Internet channel. One of the major reasons is most of the previous studies conducted analyses under a specific market condition and claimed that as the impact of Internet channel introduction. Therefore, their results are strongly influenced by the specific market settings. However, firms face various market conditions in the real worlddensity and disutility of using the Internet. The purpose of this study is to investigate the impact of various market environments on a firm's optimal channel strategy by employing a flexible game theory model. We capture various market conditions with consumer density and disutility of using the Internet.

    shows the channel structures analyzed in this study. Before the Internet channel is introduced, a monopoly manufacturer sells its products through an independent physical store. From this structure, the manufacturer could introduce its own Internet channel (MI). The independent physical store could also introduce its own Internet channel and coordinate it with the existing physical store (RI). An independent Internet retailer such as Amazon could enter this market (II). In this case, two types of independent retailers compete with each other. In this model, consumers are uniformly distributed on the two dimensional space. Consumer heterogeneity is captured by a consumer's geographical location (ci) and his disutility of using the Internet channel (${\delta}_{N_i}$).
    shows various market conditions captured by the two consumer heterogeneities.
    (a) illustrates a market with symmetric consumer distributions. The model captures explicitly the asymmetric distributions of consumer disutility in a market as well. In a market like that is represented in
    (c), the average consumer disutility of using an Internet store is relatively smaller than that of using a physical store. For example, this case represents the market in which 1) the product is suitable for Internet transactions (e.g., books) or 2) the level of E-Commerce readiness is high such as in Denmark or Finland. On the other hand, the average consumer disutility when using an Internet store is relatively greater than that of using a physical store in a market like (b). Countries like Ukraine and Bulgaria, or the market for "experience goods" such as shoes, could be examples of this market condition. summarizes the various scenarios of consumer distributions analyzed in this study. The range for disutility of using the Internet (${\delta}_{N_i}$) is held constant, while the range of consumer distribution (${\chi}_i$) varies from -25 to 25, from -50 to 50, from -100 to 100, from -150 to 150, and from -200 to 200.
    summarizes the analysis results. As the average travel cost in a market decreases while the average disutility of Internet use remains the same, average retail price, total quantity sold, physical store profit, monopoly manufacturer profit, and thus, total channel profit increase. On the other hand, the quantity sold through the Internet and the profit of the Internet store decrease with a decreasing average travel cost relative to the average disutility of Internet use. We find that a channel that has an advantage over the other kind of channel serves a larger portion of the market. In a market with a high average travel cost, in which the Internet store has a relative advantage over the physical store, for example, the Internet store becomes a mass-retailer serving a larger portion of the market. This result implies that the Internet becomes a more significant distribution channel in those markets characterized by greater geographical dispersion of buyers, or as consumers become more proficient in Internet usage. The results indicate that the degree of price discrimination also varies depending on the distribution of consumer disutility in a market. The manufacturer in a market in which the average travel cost is higher than the average disutility of using the Internet has a stronger incentive for price discrimination than the manufacturer in a market where the average travel cost is relatively lower. We also find that the manufacturer has a stronger incentive to maintain a high price level when the average travel cost in a market is relatively low. Additionally, the retail competition effect due to Internet channel introduction strengthens as average travel cost in a market decreases. This result indicates that a manufacturer's channel power relative to that of the independent physical retailer becomes stronger with a decreasing average travel cost. This implication is counter-intuitive, because it is widely believed that the negative impact of Internet channel introduction on a competing physical retailer is more significant in a market like Russia, where consumers are more geographically dispersed, than in a market like Hong Kong, that has a condensed geographic distribution of consumers.
    illustrates how this happens. When mangers consider the overall impact of the Internet channel, however, they should consider not only channel power, but also sales volume. When both are considered, the introduction of the Internet channel is revealed as more harmful to a physical retailer in Russia than one in Hong Kong, because the sales volume decrease for a physical store due to Internet channel competition is much greater in Russia than in Hong Kong. The results show that manufacturer is always better off with any type of Internet store introduction. The independent physical store benefits from opening its own Internet store when the average travel cost is higher relative to the disutility of using the Internet. Under an opposite market condition, however, the independent physical retailer could be worse off when it opens its own Internet outlet and coordinates both outlets (RI). This is because the low average travel cost significantly reduces the channel power of the independent physical retailer, further aggravating the already weak channel power caused by myopic inter-channel price coordination. The results implies that channel members and policy makers should explicitly consider the factors determining the relative distributions of both kinds of consumer disutility, when they make a channel decision involving an Internet channel. These factors include the suitability of a product for Internet shopping, the level of E-Commerce readiness of a market, and the degree of geographic dispersion of consumers in a market. Despite the academic contributions and managerial implications, this study is limited in the following ways. First, a series of numerical analyses were conducted to derive equilibrium solutions due to the complex forms of demand functions. In the process, we set up V=100, ${\lambda}$=1, and ${\beta}$=0.01. Future research may change this parameter value set to check the generalizability of this study. Second, the five different scenarios for market conditions were analyzed. Future research could try different sets of parameter ranges. Finally, the model setting allows only one monopoly manufacturer in the market. Accommodating competing multiple manufacturers (brands) would generate more realistic results.

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  • Effect of Non-Agricultural Facilities on Water Quality and Contamination in Rural Area (농촌용수 수질관리를 위한 비농업시설의 영향 연구)

    • Lee, Byung-Sun;Um, Jae-Yeon;Kim, Yang-Bin;Woo, Nam-Chil;Nam, Kyoung-Phile;Lee, Jong-Min
      • Journal of Soil and Groundwater Environment
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      • v.14 no.2
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      • pp.1-9
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      • 2009
    • This study was objected to identify the effect on water quality and contamination by non-agricultural facilities in 'A' reservoir watershed located in OO city, Kyounggi-do, Korea. Ground- and stream water samples showed (Na+K)-Cl, Ca(Cl, SO$_4$) and Ca-Cl type in an illegally discharging area of sewage and a densely industrial area indicating water contamination. Stream water of an illegally discharging area of sewage had high COD, T-N and T-P. In this area, direct incoming of sewage into stream water was induced ground water system by well pumping, and it made a progress of ground water contaminations with those components. Groundwater of a densely industrial area showed high concentrations of T-N, NO$_3$N. From a nitrogen isotope analysis, stream water of an illegally discharging area of sewage has ${\delta}^{15}N-NO_3$values of 0.7%0 was strongly affected by nitrogen originated from agrochemicals, and a densely industrial area of 19.7%0 from septic system. Ground- and stream water of a livestock fanning area were contaminated with NH$_3$-N and Mn, which was affected by intensive livestock facilities. SAR-conductivity plot indicates the water does not pose either alkalinity or salinity hazard for irrigation. COD, T-N, T-P, NO$3$-N, NH$_3$N and Mn concentrations from contaminated areas were diminished by mixing with 'A' reservoir water. There were no water contaminations in silver towns, vacationlands around reservoir and golf links. Consequently, it should be made a plan of systematic managements for past and- present possible contaminants and sewage systems in preventing water contamination by non-agricultural facilities.

    Study of Surfactant Enhanced Remediation Methods for Organic Pollutant(NAPL) Distributed over the Heterogeneous Medium (계면활성제를 이용한 불균질 매질에서 유기오염물(NAPL)의 정화효율에 관한 실험)

    • 서형기;이민희;정상용
      • Journal of Soil and Groundwater Environment
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      • v.6 no.4
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      • pp.51-59
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      • 2001
    • Column and box tests were performed to investigate the removal efficiency of NAPL using the surfactant enhanced flushing In heterogeneous medium. Homogeneous Ottawa sand and heterogeneous soil were used to verify the increase of remediation efficiency for the surfactant enhanced flushing in column test. Box tests with two different heterogeneous sub-structure were performed to quantify the capability of the surfactant enhanced flushing as a remediation method to remove NAPL from the heterogeneous medium. Two different grain size sand layers were repeated in the box to simulate the heterogeneous layer formation and the modified fault structure was built to simulate the fault system in the box. O-xylene as a LNAPL and PCE as a DNAPL were used and oleamide as a non-ionic surfactant. The maximum NAPL effluent concentration with 1% oleamide flushing in the homogeneous column test increased about 460 times compared to that with only water flushing and about 250 times increased in the real soil column test. In heterogeneous medium, the maximum effluent concentration increased about 150 times in 1% oleamide flushing and most of NAPL were removed from the box within 8 pore volume flushing, suggesting that the removal efficiency increased very much compared to in only water flushing. Results investigated the capability of the surfactant enhanced remediation method to remove NAPL even in heterogeneous medium.

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    Genetic Environments of Hydrothermal Vein Deposits in the Pacitan District, East Java, Indonesia (인도네시아 동부자바 빠찌딴(Pacitan) 광화대 열수 맥상 광상의 성인 연구)

    • Choi, Seon-Gyu;So, Chil-Sup;Choi, Sang-Hoon;Han, Jin-Kyun
      • Economic and Environmental Geology
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      • v.28 no.2
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      • pp.109-121
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      • 1995
    • The hydrothermal vein type deposits which comprise the Kasihan, Jompong and Gempol mineralized areas are primarily copper and zinc deposits, but they are also associated with lead and/or gold mineralization. The deposits occur within the Tertiary sedimentary and volcanic rocks in the Southern Mountain zone of the eastern Java island, Indonesia. Mineralization can be separated into two or three distinct stages (pre-and/or post- ore mineralization stages and main ore mineralization stage) which took place mainly along pre-existing fault breccia zones. The main phase of mineralization (the main ore stage) can be usually classified into three substages (early, middle and late) according to ore mineral assemblages, paragenesis, textures and their chemical compositions. Ore mineralogy and paragenesis of the three areas in the district are different from each other. Pyrite, pyrrhotite (/arsenopyrite), iron-rich (up to 20.5 mole % FeS) sphalerite and (Cu-)Pb-Bi sulfosalts are characteristic of the deposits in the Kasihan (/Jompong) area. On the other hand, pyrite + hematite + magnetite + iron-poor (2.7 to 3.6 mole % FeS) sphalerite assemblage is restricted to the Gempol area. Fluid inclusion data suggest that fluids of the main ore stage evolved from initial high temperatures (near $350^{\circ}C$) to later lower temperatures (near $200^{\circ}C$) with salinities ranging from 0.8 to 10.1 equiv. wt. percent NaCl. Each area represents a separate hydrothermal system: the mineralization at Kasihan and Jompong were largely due to early fluid boiling coupled with later cooling and dilution, whereas the mineralization at Gempol was mainly resulted from cooling and dilution by an influx of cooler meteoric waters. Fluid inclusion evidence of boiling indicates that pressures of ${\geq}95$ to 255 bars (${\geq}95$ bars for the Gempol area: $\approx$ 120 to 170 bars for the Jompong area: $\approx$ 140 to 255 bars for the Kasihan area) during portions of main ore stage mineralization. Equilibrium thermodynamic interpretation indicates that the evolution trends of the temperature versus fS2 variation of ore stage fluids in the Pacitan district follow two fashions: ore fluids at Kasihan and Jompong changed from the pyrite-pyrrhotite sulfidation stage towards pyritehematite- magnetite state, whereas those at Gempol evolved nearly along pyrite-hematite-magnetite reaction curve with decreasing temperature. The sulfur isotope compositions of sulfide minerals are consistent with an igneous source of sulfur with a ${\delta}^{34}S_{{\Sigma}s}$ value of about 3.3 per mil. The oxygen and hydrogen isotopic compositions of the fluids in each area indicate a progressive shift from the dominance of highly exchanged meteoric water at early hydrothermal systems towards an un- or less-exchanged meteoric water at later hydrothermal systems.

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    Quality Characteristics during Storage of Ginseng Washed by Different Methods (세척방법에 따른 인삼의 저장 중 품질특성)

    • Lee, Hyun-Seok;Cha, Hwan-Soo;Kim, Byeong-Sam;Kwon, Ki-Hyun
      • Food Science and Preservation
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      • v.16 no.3
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      • pp.342-347
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      • 2009
    • We sought to improve the methods for washing fresh raw ginseng. The quality of ginseng surface-washed by different methods was evaluated during storage at 10C and $20^{\circ}C$. The raw ginseng surface-washing method was a full-cone spray-type procedure using water and air. The water for decontamination had an electrolysis value of 80 ppm, also known as electrolysis water $2^{\circ}C$ water and water containing 5 ppm chlorine dioxide, were also used for decontamination. The Hunter color (${\Delta}E$) of ginseng washed with water withan electrolysis value of 80 ppm, or water with 5 ppm chlorine dioxide, was greater than that seen after other washing methods were used. The weight loss after washing with 5 ppm chlorine dioxide water was similar to that seen after washing with $2^{\circ}C$ water or 80 ppm electrolysis water. Reductions in total microorganism levels, and counts of yeasts and molds, assayed 10 days after washing with 5 ppm chlorine dioxide water were greater than seen after use of other sterilization methods. Quality maintenance on storage, at both 10C and 20C, after washing with 80 ppm electrolysis water, was better than that noted after other sterilization methods. The moisture content of washed ginseng was similar under all storage conditions tested.

    Studies on Processing ana Keeping Quality of Retort Pouched Foods (3) Preparation and Keeping Quality of Retort Pouched Fried Mackerel Meat Paste (레토르트파우치식품의 가공 및 품질안정성에 관한 연구 (3) 레토르트파우치 튀김어묵의 제조 및 저장중의 품질안정성)

    • LEE Eung-Ho;OH Kwang-Soo;KOO Jae-Geun;PARK Hyang-Suk;CHO Soon-Yeong;CHA Yong-Jun
      • Korean Journal of Fisheries and Aquatic Sciences
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      • v.17 no.5
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      • pp.373-382
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      • 1984
    • Processing conditions of retort pouched fried mackerel fish meat paste and quality stability during storage were investigated. The reasonable amounts of added ingredients to the frozen mackerel meat paste were $10\%$ of corn starch, $1\%$ of soybean protein, $1.5\%$ of sodium chloride, $0.6\%$ of monosodium glutamate, $0.3\%$ of alcoholic extract of red pepper, and $0.1\%$ of sodium erythorbate as an antioxidant and also added water corresponding to $10\%$ of the frozen mackerel meat paste. After grinding the defrosted mackerel fish meat paste with ingredients, the meat paste was molded in bar type and fried in soybean oil at $170-180^{\circ}C$ for 3 minutes. The fried mackerel meat paste was cooled, vacuum-packed in laminated plastic film bag (polyester/polyvinylidene chloride/unoriented polypropylene : $12{\mu}m/15{\mu}/50{\mu}m,\;14{\times}19cm$) and finally sterilized at $120^{\circ}C$ for 20 minutes in a hot water circulating retort. The pH, volatile basic nitrogen, moisture content, water activity, color, thiobarbituric acid value, peroxide value, texture and viable bacterial count of products were examined during 100 days of storage at $25{\pm}3^{\circ}C\;and\;5^{\circ}C$. The results showed that products could be preserved in good condition for 100 days at $25{\pm}3^{\circ}C$. Judging from sensory evaluation, the quality of products was not inferior to that of market products.

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    Physicochemical Properties of Diverse Rice Species (품종별 쌀의 이화학적 특성)

    • Choi, Ok-Ja;Kim, Yong-Doo;Shim, Jae-Han;Noh, Myeong-Hee;Shim, Ki-Hoon
      • Food Science and Preservation
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      • v.19 no.4
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      • pp.532-538
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      • 2012
    • Seven rice varieties (Dasan, Keunseon, Goami, Baekjinju, Seolgaeng, Hangangchal and Heukseol) were used to study the physicochemical properties of dry milled (200-mesh) rice flour species. The moisture and crude protein contents of rice were 15.00-15.10% and 6.09-8.21%, respectively. The crude lipid and crude ash of rice were 0.21-1.02% and 0.37-1.62%, respectively. As for the Hunter's color value, the L value was highest in the Dasan flour (96.47); the a value was highest in the Heukseol flour (5.03); and the b value was highest in the Baekjinju flour (3.36). The water aborption index was highest in the Goami flour (1.45), and the water solubility index was highest in the Hangangchal flour (9.16%). The amylose contents of the rice flour species were highest in the Goami (26.42%) rice flour, followed by the Dasan (19.39%), Seolgaeng (19.24%), Keunseon (18.06%), Heukseol (15.52%), Baekjinju (9.16%), and Hangangchal (0.84%) rice flour. In the X-raydiffractin patterns of the diverse species, seven tice varieties showed A-type crystallinity. As for the amylogram properties, the initial pasting temperature was 58.00-$69.03^{\circ}C$. The maximum viscosity was highest in the Dasan flour. The Heukseol flour had the lowest maximum viscosity, breakdown, and setback. In terms of the thermal properties of the differential scanning calorimeter (DSC), the onset temperature was 59.03-$66.84^{\circ}C$; the peak temperature, 66-70-$72.82^{\circ}C$; and the end temperature, 74.06-$78.66^{\circ}C$. The enthalpy (${\Delta}H$) was lowest in the Heukseol flour (7.59 J/g) and highest in the Seolgaeng flour (11.36J/g).