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Studies on the Microflora and Enzymes Influencing on Korea Native Kochuzang (Red Pepper Soybean Paste) Aging (재래식(在來式) 고추장 숙성(熟成)에 미치는 미생물(微生物) 및 그 효소(酵素)에 관(關)한 연구(硏究))

  • Lee, Ke-Ho;Lee, Myo-Sook;Park, Sung-O
    • Applied Biological Chemistry
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    • v.19 no.2
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    • pp.82-92
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    • 1976
  • The study was carried out to investigate the changes of the various chemical components and the microflora during the aging period of Korean navive Kochuzang. (Red pepper soybean paste) Korean native maeju loaves were separated into surface and inner parts. Three kinds of Korean native Kochuzang were prepared from surface part, inner part, and ordinary of maeju. The selection and the indentification of the high enzyme producing strains from the microflora and characteristics of their enzymes were studied. I. The changes of the various chemical components during the aging period of Kochuzang. 1) The changes of pH in the 3 kinds of Kochuzang displayed rapid decrease for the first 10 days after preparing and gradual curve of decrease until 60 days, but slight increase for the next 30 days. The pH of the surface part Kochuzang was lower than that of inner part or ordinary Kochuzang. 2) The total acid contents in the 3 kinds of Kochuzang showed gradual increase until the 60 days but it slowly reduced after this time. 3) The total nitrogen contents in the 3 kind of Kochuzang showed gradual inerease up to the 60 days, but slight decrease after this time. 4) The changes of trichloroacetic acid soluble nitrogen in the 3 kinds of Kochuzang showed a remarkable increase for the first 10 days, however gradual increase after this time. 5) The increase of amino nitrogen contents in the 3 kinds of Kochuzang seemed to be remarkable until the first 30 days, however to be less remarkable after this time. 6) The contents of reducing sugar in the 3 kinds of Kochuzang showed remarkable increase until the first 50 days and it slowly reduced after this time. II. The changes of microflora during the aging period of Kochuzang. 1) Aerobic, anaerobic bacteria and mold in the 3 kinds of Kochuzang were increased until the first 30 to 40 days, but they were reduced after this time. 2) No yeast in the three kinds of Kochuzang appeared until the first 20 days. Yeast were proved to grow, when the pH value was decreased below 5.4 after the 30 days. Yeasts in the surface part and ordinary Kochuzang were gradually increased and those in the inner part Kochuzang were decreased as aging. III. The selection and identification of high amylase and protease producing strains from the microflora during the aging period of Kochuzang. 1) The amylase and protease highly producing strains from microflora were identified as Bacillus subtilis-P, Bacillus subtilis-G, Bacillus licheniformis-K, Aspergillus oryzae-B. 2) Amylase activity of Aspergillus oryzae-B was highest among the strains and the strains in order of the higher activity to the lower one were Bacillus subtilis-P Bacillus licheniformis-K, Bacillus subtilis-G. Protease activities of Aspergillus oryzae-B and Bacillus subtilis-P were about the same and the strains in order of the higher activity to the lower one were Bacillus licheniformis-K, Bacillus subtilis-G. 3) Amylase activity was inhibited more than protease activity was with NaCl concentration. Amylase activity was inhibited by 45 to 65 percent and protease activity by 40 to 46 percent at the concentration of 15 percent NaCl, which was the average concentration of NaCl in Kochuzang.

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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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Effects on School Lunch Service Programme of Elementary School in Rural Area (농촌지역(農村地域) 국민학교(國民學校) 급식아동(給食兒童)과 성장발달(成長發達)과 식생활(食生活) 습관(習慣))

  • Park, Jin Wook;Lee, Sung Kook
    • Journal of the Korean Society of School Health
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    • v.5 no.2
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    • pp.74-90
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    • 1992
  • The purpose of this study is to know the effects on school lunch service programme of elementary school in rural area, by using the group consisting of the sixth year students in the schools that have provided them with the lunch for six years or longer(male student:312, & female student:324), while using the comparing group consisting of the sixth year students in the schools that have not provided them with the school lunch under their similar living condition(male student: 306 & female student:322). In addition, this study was carried out by examining all continued information about their height and weight shown in the developmetal record for six years from the 1st to 6th year, and by checking their eating habits on the basis of questionnaires. The result of this study is summarized as follows. As the result of comparing the values of their height and weight grown for 6 years, it was shown that the height of the male group provided with school lunch is 27.8 cm while the male group without lunch is 27.1 cm. And the female group provided with school lunch indicated the growing value of 29.9 cm while the group without lunch did 28.4 cm. Then, it appeared that both male and female groups provided with school lunch show higher growing values of 0.7 cm, respectively, and 1.5 cm than these groups without lunch. Also, the weight of the group without lunch was 14.8 kg. Moreover, the weight of the female group provided with school lunch was 16.9 kg while the group without lunch was 17.2 kg. Then, it was shown that the male group provided with school lunch indicates heavier growing value of 0.9 kg than the group without lunch while the female group without lunch does heavier value of 0.3 kg than the group provided with school lunch. It's figure showed that although this distribution according to percentile in the 1st year is similar to the standard regular curve it is positioned in the upper group(more thatn 70%) divided centering around 50% in the 6th year, of which distribution of children provided with school lunch was higher. When comparing the values of physical status in the 6th year, it was also shown that male children with school lunch are better than these children without lunch in jumping, throwing, chinning and lifting while female children are better than these children without lunch only in jumping, which were a significant difference. In addition, the group provided with lunch showed distribution of the higher physical grade. The result of analysis on their breakfast indicated that the children with every morning breakfast account for 67.6% of the group provided with school lunch while the group without lunch for 57.8%. Regarding the reason that they do not have the breakfast, the group with school lunch answered "Because of habits"(50.7%) while the group without lunch did "Because they have no appetite"(58.9%). When comparing the degree of preference for hot or salty food, it was apparent that these children with school lunch generally tend to prefer less hot or sailty food. With respect to the frequency and place of their eating between meals, it was shown that about 70.0% of both groups has the eating between meals, more than one time a day. Then, the group with school lunch had the eating between meals at home(45.2%) while the group without lunch did it in the process of returning to home(48.4%). Regarding the degree of their preference for a certain food, it was shown that more children of the group with school lunch do not prefer a food to others. Also, their eating attitude indicated that such children as eating the food with chat after completely swallowing food and with T.V watching are larger and lower among the group with school lunch, which showed a remarkable defference from the group without lunch. With respect to their sanitary habits such as hand washing and toothing, these children who always wash their hand before eating, accounted for 84.4 % of the group provided with school lunch while the group without lunch did for 63.6%, of which the female group with school lunch indicated a remarkable difference. The actual condition of their nutrition education showed that these children who answered "Received this education" accounted for 78.0% of the group with school lunch while the group without lunch accounted for 57.5%.

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A Study on the Compositional Characteristics of Water Systems and Landscapes in Traditional Chinese Seowons (중국 전통서원의 수체계와 수경관의 구성적 특성)

  • MA, Shuxiao;RHO, Jaehyun
    • Korean Journal of Heritage: History & Science
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    • v.55 no.3
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    • pp.74-100
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    • 2022
  • The purpose of this study was to investigate the characteristics of Chinese seowons and to obtain data based on the characteristics of waterscapes unique to Korean seowons. The conclusion of this study from the results of investigation and analysis of the location, water system, and design characteristics of 10 representative traditional seowons in China including Yuelu Seowon(嶽麓書院) conducted based on literature research and field observation is as follows. The water system of Chinese seowons is dualized into an inner and an outer water system, and in general, two and a maximum of three water bodies are superimposed on the outside. The locations of seowons are classified into five types: Four double-sided round water type sites, three converted face water type sites, one three-sided round water type site, a four-sided round water type, and a waterproofing type(依山傍水型). Therefore, compared to the typical Korean seowon facing water in the front and a mountain in the back(背山面水型), the Chinese seowons showed a highly hydrophilic property. The water shapes of the external water system were meandering(46.0%), mooring(36.0%), and broad and irregular(9.0%). In addition, water conception(水態) were streams(31.8%), rivers(27.3%), springs(13.6%), falls(9.1%), lakes(4.5%) and ponds(4.5%), in that order. As for waterscapes of the water system inside the seowon, there were seven in Akrok Seowon and four in Mansong Seowon, indicating a comparatively higher number of waterscapes. Since the 27 detailed waterscapes in 10 seowons that were the subject of the study were classified into six types including ponds and half-moon ponds, they appeared to be more diverse than the Korean seowon. It is noteworthy that in the interior waterscape of the traditional Chinese seowon, the ritualistic order, where at least one half-moon pond or square pond(方池) was arranged, is well displayed. In particular, the half-moon pond(伴池), which is difficult to find in Korean seowon, was found to be a representative waterscape element, accounting for 42.8%. If the square pond of Nanxi Seowon based on Zhu Xi's poem 「Gwanseoyugam(觀書有感)」 is also treated as a square-shaped half-moon pond, the proportion of half-moon ponds in the waterscape will be as high as 50%. The pond shapes consisted of 28% square, 24% each for free curve and round shape, 20% for semi-moon shape, and 3.8% for mountain stream type. This seems to differ greatly from the square-shaped Korean seowon. On the other hand, there were a total of 10 types of structures related to the waterscape inside the Chinese seowon: 11(26.8%) pavilion and bridge sites, five gate room sites(牌坊: 16.5%), four gate and tower sites(樓, 1.4%), two Jae sites(齋, 6.2%), and one site each(3.1%) of Heon(軒), Sa(祠), Dae(臺), and Gak(閣). In particular, the pavilions inside seowon were classified into three types: landscape pavilion(景觀亭 27.2%), tombstone pavilion(碑亭, 18.2%), and banquet pavilion(宴集亭, 54.5%). In general, it was confirmed that the half-moon pond with a pedestal bridge, and the pavilion were the major components with a high degree of connection that dominate the waterscape inside the Chinese seowon.

A Study on the Excavated Sab(a funeral fan) from Lime-filled Tomb and Lime-layered Tomb during the Joseon Dynasty (조선시대 회격·회곽묘 출토 삽(翣)에 대한 고찰)

  • Yi, Seung Hae;An, Bo Yeon
    • Korean Journal of Heritage: History & Science
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    • v.41 no.2
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    • pp.43-59
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    • 2008
  • Sap(?, a funeral fan) is a funeral ceremonial object used in association with a Confucian ceremonial custom, which was crafted by making a wooden frame, attaching a white cloth or a thick paper onto it, drawing pictures on it, and making a holder for a handle. According to Liji(Records of Rites), Sap was used since the Zhou Dynasty, and these Chinese Sap examples are no big different than the Korean Sap examples, which were described in Joseon Wangjo Sillok(Annals of the Joseon Dynasty), Gukjo Oryeui(the Five Rites of the State), and Sarye Pyeollam(Handbook on Four Rituals). This study explored Sap excavated in lime-filled tombs and lime-layered tombs of aristocrats dating back to Joseon, as well as their historical records to examine Sap's characteristics according to their examples, manufacturing methods, and use time. The number and designs of Sap varied according to the deceased' social status aristocrats used mainly one pair of 亞-shaped Bulsap, and a pair of Hwasap with a cloud design depicted on it. A Sap was wrapped twice with Chojuji paper or Jeojuji paper, and for the third time with Yeonchangji paper. Then, it was covered with a white ramie, a hemp, a cotton, a silk satin, etc. Bobul(an axe shape and 亞-shape design) was drawn on both sides of Sap, and a rising current of cloud was drawn at the peripheral area mainly with red or scarlet pigments. Sap, which were excavated from aristocrats'lime-filled and lime-layered tombs, are the type of Sap which were separated from its handle. These excavated Sap are those whose long handles were burnt during the death carriage procession, leaving Sap, which later were erected on both sides of the coffin. The manufacturing process of excavated relics can be inferred by examining them. The excavated relics are classified into those with three points and those with two points according to the number of point. Of the three-point type(Type I), there is the kind of relic that was woven into something like a basket by using a whole wood plate or cutting bamboo into flat shapes. The three-point Sap was concentrated comparatively in the early half of Joseon, and was manufactured with various methods compared with its rather unified overall shape. In the meantime, the two-point Sap was manufactured with a relatively formatted method; its body was manufactured in the form of a rectangle or a reverse trapezoid, and then its upper parts with two points hanging from them were connected, and the top surface was made into a curve(Type II) or a straight line(Type III) differentiating it from the three-point type. This manufacturing method, compared with that of the three-point type, is simple, but is not greatly different from the three-point type manufacturing method. In particular, the method of crafting the top surface into a straight line has been used until today. Of the examined 30 Sap examples, those whose production years were made known from the buried persons'death years inscribed on the tomb stones, were reexamined, indicating that type I was concentrated in the first half of the $16^{th}$ century. Type II spanned from the second half of the $16^{th}$ century to the second half of the $17^{th}$ century, and type III spanned from the first half of the $17^{th}$ century to the first half of the $18^{th}$ century. The shape of Sap is deemed to have changed from type I to type II and again from type II to type III In the $17^{th}$ century, which was a time of change, types II and III coexisted. Of the three types of Sap, types II and III re similar because they have two points; thus a noteworthy transit time is thought to have been the middle of the $16^{th}$ century. Type I compared with types II and III is thought to have required more efforts and skills in the production process, and as time passed, the shape and manufacturing methods of Sap are presumed to have been further simplified according to the principle of economy. The simplification of funeral ceremonies is presumed to have been furthered after Imjinwaeran(Japanese invasion of Joseon, 1592~1598), given that as shown in the Annals of King Seonjo, state funerals were suspended several times. In the case of Sap, simplification began from the second half of the $16^{th}$ century, and even in the $18^{th}$ century, rather than separately crafting Sap, Sap was directly drawn on the coffin cover and the coffin. However, in this simplification of form, regulations on the use of Sap specified in Liji were observed, and thus the ceremony was rationally simplified.