• Title/Summary/Keyword: Chemical extraction

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Conservation and Scientific Analysis of Human Bone Excavated in Sabi Period of Baekje from Eungpyeong-ri, Buyeo (부여 응평리 출토 백제 사비기 인골 보존처리 및 과학적 분석)

  • KIM, Mijeong;LEE, Yunseop;CHO, Eunmin;PARK, Sujin;MOON, Minseong
    • Korean Journal of Heritage: History & Science
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    • v.55 no.1
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    • pp.305-321
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    • 2022
  • The stone chamber tomb in Eungpyeong-ri, Buyeo, is a joint tomb that contains the bodies of two individuals. This paper investigates the relationship between the buried persons and the characteristics of the stone chamber tomb. Based on the geographical location, relics, and the excavated human bones, it was determined that the tomb was built during the Sabi Period of the Baekje Dynasty and that the buried individuals were most probably residents of high stature or government officials. To study the excavated bones, the remains were carefully collected and conservation was carried out. Before collecting samples from the human bones for the analytical research, the results of near-infrared analysis were used to collect the samples for the isotope analysis and DNA analysis. The most important issue when handling the excavation site was the reinforcing agent and the concentration of the agent used. In situations like this, Paraloid B-72 is the most suitable agent. When the shape of human bones was difficult to distinguish from the soil, conservation was performed using X-ray and CT imaging data. The same chemical used for the reinforcement of the site was used to complete a minimum level of conservation to the surface areas where the conservation treatment of removing foreign substances, the reinforcement areas, and bonded areas were carried out. The collagen yield from the sample obtained at selected position was 3.8% to 6.1%. The results of analyzing the stable isotopes of carbon and nitrogen found in the extracted collagen showed that the stable isotope ratios came out to δ13C -18.3‰±0.1‰, -19.0‰±0.1‰ for EBW and δ15N 10.7‰±0.5‰, 10.6‰±0.1‰ for EBE. It is believed the two individuals consumed small amounts of minor cereals, mainly from C3 plants, and protein was obtained from eating terrestrial animals. What's more, the deviations in data obtained from the two individuals were so small that it could be inferred that the individuals ate similar foods. Considering the preservation state of the sample, amplifying DNA for the DNA analysis would have been very difficult since the amount of surviving DNA was so deficient. For DNA analysis, it is anticipated that the results could be derived by applying improved extraction methods that will be developed in the future. In this research, any association between scientific analysis(DNA and stable isotope ratio) and near-infrared spectroscopy was difficult to establish. Further research is needed on the utilization of near-infrared analysis for gathering samples from human bones.

Studies on the Physical and Chemical Denatures of Cocoon Bave Sericin throughout Silk Filature Processes (제사과정 전후에서의 견사세리신의 물리화학적 성질변화에 관한 연구)

  • 남중희
    • Journal of Sericultural and Entomological Science
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    • v.16 no.1
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    • pp.21-48
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    • 1974
  • The studies were carried out to disclose the physical and chemical properties of sericin fraction obtained from silk cocoon shells and its characteristics of swelling and solubility. The following results were obtained. 1. The physical and chemical properties of sericin fraction. 1) In contrast to the easy water soluble sericin, the hard soluble sericin contains fewer amino acids include of polar side radical while the hard soluble amino acid sach as alanine and leucine were detected. 2) The easy soluble amino acids were found mainly on the outer part of the fibroin, but the hard soluble amino acids were located in the near parts to the fibroin. 3) The swelling and solubility of the sericin could be hardly assayed by the analysis of the amino acid composition, and could be considered to tee closely related to the compound of the sericin crystal and secondary structure. 4) The X-ray patterns of the cocoon filament were ring shape, but they disappeared by the degumming treatment. 5) The sericin of tussah silkworm (A. pernyi), showed stronger circular patterns in the meridian than the regular silkworm (Bombyx mori). 6) There was no pattern difference between Fraction A and B. 7) X-ray diffraction patterns of the Sericin 1, ll and 111 were similar except interference of 8.85A (side chain spacing). 8) The amino acids above 150 in molecular weight such as Cys. Tyr. Phe. His. and Arg. were not found quantitatively by the 60 minutes-hydrolysis (6N-HCI). 9) The X-ray Pattern of 4.6A had a tendency to disappear with hot-water, ether, and alcohol treatment. 10) The partial hydrolysis of sericin showed a cirucular interference (2A) on the meridian. 11) The sericin pellet after hydrolysis was considered to be peptides composed with specific amino acids. 12) The decomposing temperature of Sericin 111 was higher than that of Sericin I and II. 13) Thermogram of the inner portioned sericin of the cocoon shell had double endothermic peaks at 165$^{\circ}C$, and 245$^{\circ}C$, and its decomposing temperature was higher than that of other portioned sericin. 14) The infrared spectroscopic properties among sericin I, II, III and sericin extracted from each layer portion of the cocoon shell were similar. II. The characteristics of seriein swelling and solubility related with silk processing. 1) Fifteen minutes was required to dehydrate the free moisture of cocoon shells with centrifugal force controlled at 13${\times}$10$^4$ dyne/g at 3,000 R.P.M. B) It took 30 minutes for the sericin to show positive reaction with the Folin-Ciocaltue reagent at room temperature. 3) The measurable wave length of the visible radiation was 500-750m${\mu}$, and the highest absorbance was observed at the wave length of 650m${\mu}$. 4) The colorimetric analysis should be conducted at 650mu for low concentration (10$\mu\textrm{g}$/$m\ell$), and at 500m${\mu}$ for the higher concentration to obtain an exact analysis. 5) The absorbing curves of sericin and egg albumin at different wave lengths were similar, but the absorbance of the former was slightly higher than that of the latter. 6) The quantity of the sericin measured by the colorimetric analysis, turned out to be less than by the Kjeldahl method. 7) Both temperature and duration in the cocoon cooking process has much effect on the swelling and solubility of the cocoon shells, but the temperature was more influential than the duration of the treatment. 8) The factorial relation between the temperature and the duration of treatment of the cocoon cooking to check for siricin swelling and solubility showed that the treatment duration should be gradually increased to reach optimum swelling and solubility of sericin with low temperature(70$^{\circ}C$) . High temperature, however, showed more sharp increase. 9) The more increased temperature in the drying of fresh cocoons, the less the sericin swelling and solubility were obtained. 10) In a specific cooking duration, the heavier the cocoon shell is, the less the swelling and solubility were obtained. 11) It was considered that there are differences in swelling or solubility between the filaments of each cocoon layer. 12) Sericin swelling or solubility in the cocoon filament was decreased by the wax extraction.. 13) The ionic surface active agent accelerated the swelling and solubility of the sericin at the range of pH 6-7. 14) In the same conditions as above, the cation agent was absorbed into the sericin. 15) In case of the increase of Ca ang Mg in the reeling water, its pH value drifted toward the acidity. 16) A buffering action was observed between the sericin and the water hardness constituents in the reeling water. 17) The effect of calcium on the swelling and solubility of the sericin was more moderate than that of magnecium. 18) The solute of the water hardness constituents increased the electric conductivity in the reeling water.

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Studies on the Pulping Characteristics of Larchwood (Larix leptolepis Gordon) by Alkaline Process with Additives (첨가제(添加劑) 알칼리 법(法)에 의한 일본 잎갈 나무의 펄프화(化) 특성(特性)에 관(關)한 연구(硏究))

  • Lim, Kie-Pyo;Shin, Dong-Sho
    • Journal of the Korean Wood Science and Technology
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    • v.7 no.2
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    • pp.3-30
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    • 1979
  • Larch ($\underline{Larix}$ $\underline{leptolepis}$ GORDON), one of the major afforestation species in Korea in view of its growing stock and rate of growth, is not favored as a raw material for pulp due to its low yield of pulp and difficulties with bleaching arising from the high content of extractives in wood, and the high heartwood ratio and the active phenolics, respectively. The purpose of this study is to investigate the characteristics of firstly pulping with various additives of cellulose protector for the yield of pulp, and secondly bleaching with oxygen for chlotination-alkali extraction of five stage-sequence to reduce chlorine compounds in bleaching effluents. The kraft cooking liquor for five age groups of larchwood was 18 percent active alkali with 25 percent sulfidity and 5 : 1 liquor-to-wood ratio, and each soda liquor for sap-and heart-wood of the 15-year-old larchwood was 18 percent alkali having one of the following cellulose protectors as the additive; magnesium sulfate ($MgSO_4$, 2.5%), zinc sulfate ($ZnSO_4$, 2.5%), aluminium sulfate ($Al_2(SO_4)_3$, 2.5%), potasium iodide (KI, 2.5%), hydroquinone (HQ, 2.5%), anthraquinone (AQ, 0.1%) and ethylene diamine (EDA, 2.5%). Then each anthraquinone-soda liquor for the determination of suitable cooking condition was the active alkali level of 15, 17 and 19 percent with 1.0, 0.5 and 0.1 percent anthraquinone, respectively. The cooking procedure for the pulps was scheduled to heat to 170$^{\circ}C$ in 90 minutes and to cook 90 minutes at the maximum temperature. The anthraquinone-soda pulps from both heartwood and sapwood of 15-year-old larchwood prepared with 0.5 percent anthraquinone and 18 percent active alkali were bleached in a four-stage sequency of OCED. (O: oxygen bleaching, D: chlorine dioxide bleaching and E: alkali extraction). In the first stage oxygen in atmospheric pressure was applied to a 30 percent consistency of pulp with 0.1 percent magnesium oxide (MgO) and 3, 6, and 9 percent sodium hydroxide on oven dry base, and the bleached results were compared pulps bleached under the conventional CEDED (C: chlorination). The results in the study were summarized as follows: 1. The screened yield of larch kraft pulp did not differ from particular ages to age group, but heartwood ratio, basic density, fiber length and water-extractives contents of wood and the tear factor of the pulp increased with increasing the tree age. The total yield of the pulp decreased. 2. The yield of soda pulp with various chemicals for cellulose protection of the 15-year-old larchwood increased slightly more than that of pure soda pulp and was slightly lower than that of kraft pulp. The influence of cellulose protectors was similar to the yield of pulps from both sapwood and heartwood. The effective protectors among seven additives were KI, $MgSO_4$ and AQ, for which the yields of screened pulp was as high as that of kraft pulp. Considering the additive level of protector, the AQ was the most effective in improving the yield and the quality of pulp. 3. When the amount of AQ increased in soda cooking, the yield and the quality of the pulp increased but rejects in total yield increased with decreasing the amount of active alkali from 19 to 15 percent. The best proportion of the AQ seemed to be 0.5 percent at 17 percent active alkali in anthraquinone-soda pulping. 4. On the bleaching of the AQ-soda pulp at 30 percent consistency with oxygen of atomospheric pressure in the first stage of the ODED sequence, the more caustic soda added, the brighter bleached pulp was obtained, but more lignin-selective bleaching reagent in proportion to the oxygen was necessary to maintain the increased yield with the addition of anthraquinone. 5. In conclusion, the suitable pulping condition for larchwood to improve the yield and quality of the chemical pulp to the level for kraft pulp from conventional process seemed to be. A) the selection of young larchwood to prevent decreasing in yield and quality due to the accumulation extractives in old wood, B) the application of 0.5 percent anthraquinone to the conventional soda cooking of 18 percent active alkali, and followed, C) the bleaching of oxygen in atmospheric pressure on high consistency (30%) with 0.1 percent magnesium oxide in the first stage of the ODED sequence to reduce the content of chlorine compounds in effluent.

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Studies on Fine Spirits Aging [Part I]-On the Aptitude of the Korean Oak Varieties as Barrels for Aging Apple Fine Spirits- (증류주(蒸溜酒) 숙성(熟成)에 관(關)한 연구(硏究) 제1보[第一報]-사과 증류주(蒸溜酒) 숙성(熟成)에 있어서 숙성통재(熟成桶材)로서 한국산(韓國産) 참나무 품종별(品種別) 이용적성(利用適性)에 관(關)하여-)

  • Lee, Ke-Ho
    • Applied Biological Chemistry
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    • v.20 no.1
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    • pp.66-80
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    • 1977
  • This research was carried as a part of the basic study, in which the aptitude of theKorean oak varieties as barrels for aging apple fine spirits was investigated, and thefollowing results were obtained. 1. Following was the result of the chemical analysis of the fruits which are now mass-produced and can be used as a substitute for raw materials for wine production. Apple (Malus pumila Miller var. domestica Schneider) : Total sugar. total acid, volatile acid and pectin of Jonathan (Hong-og) were 13.95%, 0.46%, 0.012%, 0.20% respectively. Total sugar, total acid, volatile acid and pectin of Ralls (Koog-kwang) were 13.35%, 0.43%, 0.011%, 0.45% respectively. 2. Because of low yield of apple juice due to cellulose, pectin, hemicellulose which are present besides sugars, acids in apples, the apple juice were treated with xylanase of Aspergillus niger SUAFM-430, cellulase and pectinase of Aspergillus niger SUAFM-6. This treatment increased the yield of apple juice. And the apple juice was sterilized by adding potassium metabisulfite $(K_2S_20_5)$ and Saccharomyces cerevisae var. ellipsoideus Rasse Johannisberg II (SUAFM-1018) as a cultivation yeast, which has a strong fermentation power was used to ferment. The yield of apple wine based on raw material was 86-87%. The amount of ethanol, extract and methanol obtained from Jonathan and Ralls were 13.5%, 5.4%, 0.04-0.05% respectively. 3. Wines were distilled for two times by the pot still method to make fine spirits. The yield of fine spirits from apple wine mash was 86.6%, and the pH of fine spirits from Jonathan and Ralls were 4.1, 4.2 respectively. 4. The oak chips made of inner part or outer part of 24 Korean oak varieties were used to select the barrel for aging fine spirits. Two oak chips (one oak chip: $1{\times}1{\times}5cm$) of the inner part or of the outer part of each oak variety were dipped into 300 ml of fine spirits, which was bottled in 640ml beer bottle, and followed aging. The colors, flavors and tastes of the fine spirits were checked during 6 months. A. As a criterion for the first screening of oak barrels for aging fine spirits, the rate five of color extraction was determined. The oak chips showed good results in their order as follows and the best 5 varieties were selected. Gal-cham: Quercus aliena Blume (Inner part), Gul-cham: Quercus variabilis Blume (Outer part), Gal-chain: Quercus aliena Blume (Outer part), Jol-cham: Quercus serrata Thumb (Inner and Outer part). Sin-gal-cham: Quercus mongolica Fisher (Outer and Inner part) Sang-su-ri: Quercus acutissima Carruthers (Outer and Inner part) B. To find out the influence of aging temperature on aging, apple fine spirits were aged by dipping each oak chip at room temperature $(24-25^{\circ}C)$) and $45^{\circ}C$. Aging at $45^{\circ}C$ gave the best result followed aging at $30^{\circ}C$ and then at room temperature. C. Apple fine spirits was aged for six months by dipping oak chips in Erlenmeyer flasks and was irradiated with U.V light. The U.V irradiation enhanced the aging effect by nearly two times, compared with the aging without U.V irradiation. D. In aging apple fine spirits by dipping two oak chips, it was observed that the extent of the extraction of most components of oak chips were strongly dependent upon the pH of fine spirits. E. Oak chips of five selected oak varieties and a Limousin white oak from France as a control were used. Each apple fine spitits was dipped by two oak chips, and was aged at room temperature $(24-25^{\circ}C)$, $30^{\circ}C$, $45^{\circ}C$, and with the U.V irradiation at room temperature shaking every week. After six months of aging, the panel test of these aged fine spirits (Young Brandy) showed the following result. Young brandy of apples aged at $45^{\circ}C$ by dipping oak chips of Gal-chain was almost as the fine spirits which were aged at room temperature by dipping Limousin white oak chips from France. Young brandy of with U.V. irradiation at room temperature which were aged by dipping oak chips of Gal-chain was a little worse than that from the fine spirits aged at room temperature by dipping Limousin white oak chips from France. And so, Korean oak varieties are thought to be able to be used for aging every apple fine spirit which was here investigated.

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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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