• Title/Summary/Keyword: separation properties.

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Purification and Biological Activity of Ecdysterone from Korean Achyranthes radix (韓國産 牛膝의 Ecdysterone 抽出과 그 生理活性에 관한 硏究)

  • Kim, Jeong-Il;Lee, Jae-Yong;Kim, Chun-Su;Park, Kwang-E.
    • Journal of Sericultural and Entomological Science
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    • v.25 no.1
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    • pp.1-20
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    • 1983
  • It has been known that the insect molting hormone and its analogues exist also in plant kingdom and their concentration has been found to be about 0.1~2.0% of dry matter, which is equivalent to $10^3{\sim}10^5$ times of those in insects. This study was carried out; 1) to isolate the phytoecdysones from Korean Achyranthes radix and characterize their physico-chemical properties. 2) to investigate the biological activity of this phytoecdysone on Bombyx mori larvae. The resuls were summarized as follows; 1. The extraction method of phytoecdysones was optimized by three consecutive reflux for 1hr using 200g of dried and milled radix per 1l methanol. 2. The purification from the crude extract was made by a series of steps such as precipitation of gum-type polymer with n-Butyl acetate, adsorption on technical grade silica and chromatography with neutral alumina. The conditions of each step were optimized and the resulting crude crystal was about 500mg per kg dry radix. 3. The crude crystal from the cultivated Achyranthes(Achyranthes japonia) contained ecdysterone (20-hydroxyecdysone) and inokosterone in the proportion of one to one. In order to separate these, a series of processes such as acetylation, separation by alumina column chromatography deacetylation by alcoholysis, deionization and crystallization were introduced and optimized 125mg of ecdysterone and 18mg of inokosterone per kg dry radix were thus obtained. 4. The wild Achyranthes (Achyranthes obtusifolia) radix was found to contain the ecdysterone only. A 285mg of ecdysterone was crystallized per kg dry radix. 5. Isolated ecdysterone, inodosterone and acetylated compounds were characterized by IR., UV., NMR spectroscopy, mp, TLC and densitometry. 6. Ligation experiment was undertaken to confirm the biological activity of the purified ecdysterone; the ecdysterone could induce larval-pupal metamorphosis in the ligated abdomen of 4th instar larvae injecting 0.5~1.0${\mu}g$. 7. By ecdysterone feeding experiment using artificial diet, it was elucidated that the critical time of feeding would be the first half of each instar resulting in increased weight of silk layer. 8. The ecdysterone was fed to 5th instar silkworm at the level of 1, 2, 3, 5ppm of dry feed of artificial diet containing 5% mulberry leaves for 72hrs. At 2ppm of the concentration. body weight and silk layer weight were arrived at maximum. But at higher concentrations body weight and silk layer weight decreased than the control group. At 2ppm of the concentration, body weight was increased by 12.5%. 9. Feeding 2ppm of ecdysterone at the later half of 5th instar, the duration of larvae was shortened.

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Factors Affecting Wet-Paddy Threshing Performance (탈곡기의 제작동요인이 벼의생탈곡성능에 미치는 영향)

  • 남상일;정창주;류관희
    • Journal of Biosystems Engineering
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    • v.5 no.1
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    • pp.1-14
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    • 1980
  • Threshing operation may be one of the most important processes in the paddy post-production system as far as the grain loss and labor requirement are concerned . head-feeding type threshers commercially available now in Korea originally were developed for threshing dry paddy in the range of 15 to 17 % in wet basis. However, threshing wet-paddy with the grain moisture content above 20 % has been strongly recommended, especially for new high-yielding Indica -type varieties ; (1) to reduce high grain loss incurred due to the handling operations, and (2) to prevent the quantitative and qualitative loss of milled -rice when unthreshed grains are rewetted due to the rainfall. The objective of this study were to investigate the adaptability of both a head-feeding type thresher and a throw-in type thresher to wet-paddy , and to find out the possiblilities of improving the components of these threshers threshing. Four varieties, Suweon 264 and Milyang 24 as Tongil sister line varieties, minehikari and Jinhueng as Japonica-type varieties, were used at the different levels of the moisture content of grains. Both the feed rate and the cylinder speed were varied for each material and each machine. The thresher output quality , composition of tailing return, and separating loss were analyzed from the sampels taken at each treatment. A separate experiment for measurement opf the power requirement of the head-feeding type thresher was also performed. The results are summarized as follows : 1. There was a difference in the thresher output quality between rice varieties. In case of wet-paddy threshing at 550 rpm , grains with branchlet and torn heads for the Suweon 264 were 12 % and 7 % of the total output in weight, respectively, and for the Minehikari 4.5 % and 2 % respectively. In case of dry paddy threshing , those for the Suweon 264 were 8 % and 5% , and for the Minehikari 4% and 1% respectively. However, those for the Milyang 23 , which is highly susceptable to shattering, were much lower with 1 % and 0.5% respectively, regardless of the moisture content of the paddy. Therefore, it is desirable to breed rice varieties of the same physical properties as well as to improve a thresher adaptable to all the varieties. Torn heads, which increased with the moisture content of rall the varieties except the Milyang 23 , decreased as the cylinder speed increased, but grains with branchlet didnt decrease. The damaged kernels increased with the cylinder speed. 3. The thresher output quality was not affected much by the feed rate. But grains with branchlet and torn heads increased slightly with the feed rate for the head-feeding type thresher since higher resistance lowered at the cylinder speed. 4. In order to reduce grains with branchlet and torn heads in wet-paddy threshing , it is desirable to improve the head-feeding type thresher by developing a new type of cylinder which to not give excess impact on kernels or a concave which has differenct sizes of holes at different locations along the cylinder. 5. For the head-feeding type thresher, there was a difference in separating loss between the varieties. At the cylinder speed of 600 rpm the separating losses for the Minehikari and the Suweon 264 were 1.2% and 0.6% respectively. The separating loss of the head-feeding type thresher was not affected by the moisture content of paddy while that of the Mini-aged thresher increased with the moisture content. 6. From the analysis of the tailings return , to appeared that the tailings return mechanism didn't function properly because lots of single grains and rubbishes were unnecessarily returned. 7. Adding a vibrating sieve to the head-feeding type thresher could increase the efficiency of separation. Consequently , the tailing return mechanism would function properly since unnecessary return could be educed greatly. 8. The power required for the head-feeding type thresher was not affected by the moisture content of paddy, but the average power increased linearly with the feed rate. The power also increased with the cylinder speed.

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Sorption of PAHs by Soil Humins and Effect of Soil Inorganic Matrixs (PAHs의 토양휴민과의 흡착특성 및 토양 무기물의 영향 해석)

  • Lim, Dong-Min;Lee, Seung-Sik;Shin, Hyun-Sang
    • Journal of Korean Society of Environmental Engineers
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    • v.28 no.12
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    • pp.1337-1346
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    • 2006
  • Soil humin is the insoluble fraction of humic materials and play an important roles in the irreversible sorption of hydrophobic organic contaminants onto soil particles. However, there have been limited knowledge about the sorption and chemical properties of humin due to the difficulties in its separation from the inorganic matrix(mainly clays and oxides). In this study, de-ashed soil humins($Hu_1-Hu_6$) were isolated from a soil residues(Crude Hu) after removing alkali-soluble organic fractions followed by consecutive dissolution of the mineral matrix with 2%-HF for 2 hr. The humin samples were characterized by elemental analysis and $^{13}C$ NMR spectroscopic method and their sorption-desorption behavior for 1-naphthol were investigated from aqueous solution. The results were compared one another and that with peat humin. $^{13}C$ NMR spectra features indicate that the soil humin molecules are mainly made up of aliphatic carbons(>80% in total carbon) including carbohydrate, methylene chain. Freundlich sorption parameter, n was increased from 0.538 to 0.697 and organic carbon-normalized sorption coefficient(log $K_{OC}$) values also increased from 2.43 to 2.74 as inorganic matrix of the soil humin removed by HF de-ashing. The results suggest that inorganic phase in humin plays an important, indirect role in 1-naphthol sorption and the effects on the sorption non-linearity and intensity are analyzed by comparison between the results of soil humin and peat humin. Sorption-desorption hysteresis were also observed in all the humin samples and hysteresis index(HI) at low solute concentration($C_e$=0.1 mg/L) are in order of Peat humin(2.67)>De-ashed humin(0.74)>Crude Hu(0.59).

Development and Research on a Functional Hydrolyzed Whey Protein Powder Product with Sialic Acid as a Marker Compound - II. Repeated 90-day Oral Administration Toxicity Test using Rats Administered Whey Protein Powder containing Highly Concentrated Sialic Acid (23%) produced by Enzyme Separation and Solvent Enrichment Method - (Sialic Acid를 지표성분으로 하는 유청가수분해단백분말의 기능성식품 개발연구 - II. 효소분리 용매정제로 고농도 Sialic Acid가 함유된 유청가수분해단백분말(23%)의 랫드를 이용한 90일 반복경구투여 독성시험 평가 연구 -)

  • Noh, Hye-Ji;Cho, Hyang-Hyun;Kim, Hee-Kyong;Koh, Hong-Bum
    • Journal of Dairy Science and Biotechnology
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    • v.34 no.2
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    • pp.117-135
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    • 2016
  • The present study was performed to develop a functional raw food material from hydrolyzed whey protein powder (23%-GNANA) medication containing sialic acid as a marker compound that is naturally occurring at 7% concentration in GMP (glycomacropeptide). GMP is used worldwide in foodstuffs for babies and infants and is obtained from the milk protein as safe food. While the purpose of our detailed evaluation was aimed to assess preliminary NOAEL values for and above 2,000 mg/kg/day, a clinical dose allowance for 23%-GNANA (as per characteristic of a functional health product, a highly refined test substance of 23% (v/v) sialic acid combined in GMP), at the same time we also wanted to assess the safety of GMP hydrolyzate lacking sialic acid but with identical properties as GMP. Animal safety evaluation was conducted using 23%-GNANA as the test substance, produced from hydrolyzed whey protein powder (product name: HELICOBACTROL-23; provided by Medinutrol Inc. [Korea]; composed of 23% sialic acid and GMP protein) after isolating the sialic acid using enzymes approved as food additives, with GMP as a raw material, and subsequently increasing the content of xx up to 23% through 80% (v/v) ethanol soaking and concentrating, in accordance with GLP Guideline. The animal safety evaluation mentioned above was made on the basis of toxicity in SPF Sprague-Dawley female and male rats dosed with 10 mL of the test substance diluted to 0, 1,250, 2,500, and 5,000 mg/kg directly into their stomachs for 90 d. This was determined in terms of the general symptoms and animal viability, weight and amount of feed intake, eye examination, uracrasia tests, hematological and blood biochemical disorder tests, blood coagulation test, abnormal intestine weight, abnormalities during postmortem and histopathological examinations. Statistical significance was set at P<0.05. Based on the toxicity determination, a certain minor effect associated with the test substance was observed in male rats with no major effects of the tested substance, in comparison with the control group dosed with sterilized water. Nevertheless, the NOAEL value, evaluated as per toxicity criteria, was verified as 5,000 mg/kg/day (P<0.05). Similarly, for female rats, a certain minor effect associated with the test substance was observed in 5,000 mg/kg/day dosed group, with no major effect, yet the NOAEL value (as assessed as per toxicity criteria) was determined to be 5,000 mg/kg/day (P<0.05), which was the same as for male rats. Accordingly, the NOAEL values of the test substances for all female and male rats were finally verified as 5,000 mg/kg/day (P<0.05). In conclusion, it was determined that the 23%-GNANA test substance exceeds 2,000 mg/kg/day, the clinical allowance characteristic for functional health food, and was finally evaluated to cause no safety concerns when used as a raw material in functional health food production, which was the ultimate goal of the present study.

STUDIES ON THE EXTRACTION OF SEAWEED PROTEINS 1. Extraction of Water Soluble Proteins (해조단백질의 추출에 관한 연구 1. 수용성 단백질의 추출)

  • RYU Hong-Soo;LEE Kang-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.10 no.3
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    • pp.151-162
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    • 1977
  • Distribution of marine algae is diverse in Korea and the resource of edible algae is abundant marking 239,037 tons of yearly production in 1976. They have been known as a protein source and used as a supplement in Korean diet. It is necessary to estimate the potentiality and properties of usable algal proteins especially as food resources and studies of extraction and separation of the proteins, therefore, are basically required for this purpose. In this study, the influence of various factors including the sample treatment, extraction time and temperature, sample us extraction solvent ratio and pH upon the extractability of the water soluble protein was determined. And the effect of precipitation treatment for isolation of the algal protein from the extracts was also tested. Nine species of algae, the major ones in consumption as food namely Porphyra suborbiculata, Undaria pinnatifida, Hizikia fusiforme, Sargassum fulvellu, Enteromorpha linza, Codium fragile, Sargassum kjellmanianum and Ulva pertusa were collected as fresh from Kijang, Yangsan Gun, in the vicinity of Busan city. The content of crude protein $(N\times6.25)$ of the algae ranged from $9.46\%\;to\;24.14\% showing the highest value in Porphyra suborbiculata and the minimum in Hizikia fusiforme. In the effort of maceration of blending methods on the extractability, immersion freezing in dry ice-methanol solution appeared most effective yielding 1.5 to 2.5 times extractability than that of the mortar grinding method. The effect of the ratio of sample vs solvent on extractability differed from species. It was enhanced at the ratio of 1:20 (w/v) in Ulva pertusa and Enteromorpha linza while the ratio was 1:30 (w/v) for Cedium fragile, Undaria pinnatifida, Hizikia fusiferme, Sargassum fulvellum and Porphyra suborbiculata and 1:40 for Sargassum kjellmanianum respectively. The effect of extraction time and temperature was revealed differently from species which might be caused by differences in the constitution of algal tissues resulting in that the extraction for 1 hour at $50^{\circ}C$ gave the maximum extractabilily in Ulva pertusa and Enteromorpha linza, 2 hours in Porphyra suborbiculata, Hikikia fusiforme, Undaria pinnatifida, Sargassum kjellmanianum and 3 hours in Codium fragile. And the extractability was higher at $50^{\circ}C$ to $60^{\circ}C$ for the most of the tested samples except Hizikia fusiforme. The optimum pH for the extraction was 9 to 12. The recovery of extractable nitrogen to the total nitrogen was $63\%$ in average with the first extracts and $8.6\%$ with the second extracts respectively. Both extracts were prepared by 2 hour extraction at $50{\pm}1^{\circ}C$ with dry ice-methanol frozen and seasand macerated materials. And these conditions assumed to be an optimum for the extraction of water soluble algal proteins since the nitrogen content after the first extraction covered $90\%$ of the total water extractable nitrogen. In the precipitation of the extracted proteins, Barnstein method and methanol treatment seemed to be more efficient than other precipitation methods.

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Gas Permeation Properties of Sulfonated 6FDA-Based Polyimide Membranes Exchanged with Metal Ions (금속이온이 치환된 설폰화된 6FDA계 폴리이미드 막의 기체 투과 특성)

  • Im, Hyeon-Soo;Lee, Byung-Seong;Lee, Bo-Sung;Yoon, Seok-Won;Koh, Hyung-Chul;Lee, Choong-Sub;Ha, Seong-Yong;Cheong, Seong-Ihl;Rhim, Ji-Won
    • Polymer(Korea)
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    • v.33 no.6
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    • pp.555-560
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    • 2009
  • Sulfonic acid of the sulfonated 6FDA-based polyimides were exchanged with the monovalent ($Li^+$, $Na^+$, $K^+$) and divalent ($Mg^{2+}$, $Ca^{2+}$, $Ba^{2+}$) ions. The effect of metal cations exchanged sulfonated polyimides was investigated in terms of gas permeability and selectivity for $CO_2$, $O_2$ and $N_2$ gases. Thermogravimetric analysis showed that thermal stability of sulfonated polyimide was improved by exchanged metal cations. The permeabilities of monovalent cation-exchanged, sulfonated polyimide were reduced as the ion radius reduced [$Li^+$(0.059 nm)>$Na^+$(0.102 nm)>$K^+$(0.138 nm)], and those of divalent cations exchanged were determined by the ionic radii and electrostatic crosslinking between the polymer and metal cations, whereas the selectivities of all the metal cation-exchanged, sulfonated polyimides for $CO_2/N_2$ and $O_2/N_2$, were higher than those of sulfonated polyimide membranes. The sulfonated polyimide exchanged with the potassium cation showed the $O_2$ permeability of 89.98 Barrer [$1\times10^{-10}\;cm^3$(STP) $cm/cm^2{\cdot}s{\cdot}cmHg$] and the sulfonated polyimide exchanged with the lithium cation showed the $O_2/N_2$ selectivity of 12.9.

Separation of Reducing Sugars from Rape Stalk by Acid Hydrolysis and Fabrication of Fuel Pellets from its Residues (산가수분해한 유채대로부터 유리당의 분리 및 이의 잔사로부터 펠릿의 제조)

  • Yang, In;Ahn, Byoung Jun;Kim, Myeong-Yong;Oh, Sei Chang;Ahn, Sye Hee;Choi, In-Gyu;Kim, Yong-Hyun;Han, Gyu-Seong
    • Korean Journal of Plant Resources
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    • v.27 no.1
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    • pp.60-71
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    • 2014
  • This study was conducted to identify the potential of rape stalk as a raw material for biorefinery process of rape flower. At first, rape stalk (RS) was immersed in distilled water (DW), acetic acid (AA), oxalic acid (OA), sulfuric acid (SA) and sodium hydroxide (SH) solutions, and the content of reducing sugars liberated from immersed RS was analyzed. Glucose, xylose, arabinose and sucrose were detected varying with the immersion type. In particular, 1% AA-immersion of RS for 72 hr was the most effective conditions to liberate glucose from RS. Secondly, the RS residues were used for elementary analysis and fabrication of fuel pellets. In addition to the solution type, concentration of immersion solutions (0%, 1%, 2%) and immersion time (24, 72, 120 hr) were used as experimental factors. The contents of nitrogen, sulfur and chlorine reduced effectively through the immersion of RS in DW, AA and OA solutions. For properties of RS-based pellets, bulk density and higher heating value of RS-based pellets greatly increased with the immersion of RS, and the qualities were much higher than those of the A-grade pellet of the EN standards. Ash content decreased remarkably through the immersion of RS, and was satisfied with the A-grade pellet standard. Durability was negatively affected by the immersion of RS, and did not reached to B-grade of the EN standard. In conclusion, acid immersion of RS can be a pretreatment method for the production of fuel pellet and bioethanol, but use of the immersed RS for the production of high-quality pellets might be restricted due to low durability of immersed-RS pellets. Therefore, further studies, such as investigation of detailed immersion conditions, fabrication of mixed pellets with wooden materials and addition of binders, are needed to resolve the problems.

Evaluation of microplastic in the inflow of municipal wastewater treatment plant according to pretreatment methods (전처리 방법에 따른 하수처리장 유입수에서의 미세플라스틱 성상분석 평가)

  • Kim, Sungryul;Gil, Kyungik
    • Journal of Wetlands Research
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    • v.24 no.2
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    • pp.83-92
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    • 2022
  • The amount of the plastic waste has been increasing according to global demand for plastic. Microplastics are the most hazardous among all plastic pollutants due to their toxicity and unknown physicochemical properties. This study investigates the optimal methodology that can be applied to sewage samples for detecting microplastics before discussing reducing microplastics in MWTPs. In this study, the effect of different pretreatment methods while detecting microplastic analysis of MWTP influent samples was investigated; the samples were collected from the J sewage treatment plant. There are many pretreatment methods but two of them are widely used: Fenton digestion and hydrogen peroxide oxidation. Although there are many pretreatment methods that can be applied to investigate microplastics, the most widely used methods for sewage treatment plant samples are Fenton digestion and H2O2 oxidation. For each pretreatment method, there were factors that could cause an error in the measurement. To overcome this, in the case of the Fenton digestion pretreatment, it is recommended to proceed with the analysis by filtration instead of the density separation method. In the case of the H2O2 oxidation method, the process of washing with distilled water after the reaction is recommended. As a result of the analysis, the concentration of microplastics was measured to be 2.75ea/L for the sample using the H2O2 oxidation method and 3.2ea/L for the sample using the Fenton oxidation method, and most of them were present in the form of fibers. In addition, it is difficult to guarantee the reliability of measurement results from quantitative analysis performed via microscope with eyes. A calibration curve was created for prove the reliability. A total of three calibration curves were drawn, and as a result of analysis of the calibration curves, all R2 values were more than 0.9. This ensures high reliability for quantitative analysis. The qualitative analysis could determine the series of microplastics flowing into the MWTP, but could not confirm the chemical composition of each microplastic. This study can be used to confirm the chemical composition of microplastics introduced into MWTP in the future research.

A Study on the Resource Recovery of Fe-Clinker generated in the Recycling Process of Electric Arc Furnace Dust (전기로 제강분진의 재활용과정에서 발생되는 Fe-Clinker의 자원화에 관한 연구)

  • Jae-hong Yoon;Chi-hyun Yoon;Hirofumi Sugimoto;Akio Honjo
    • Resources Recycling
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    • v.32 no.1
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    • pp.50-59
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    • 2023
  • The amount of dust generated during the dissolution of scrap in an electric arc furnace is approximately 1.5% of the scrap metal input, and it is primarily collected in a bag filter. Electric arc furnace dust primarily consists of zinc and ion. The processing of zinc starts with its conversion into pellet form by the addition of a carbon-based reducing agent(coke, anthracite) and limestone (C/S control). These pellets then undergo reduction, volatilization, and re-oxidation in rotary kiln or RHF reactor to recover crude zinc oxide (60%w/w). Next, iron is discharged from the electric arc furnace dust as a solid called Fe clinker (secondary by-product of the Fe-base). Several methods are then used to treat the Fe clinker, which vary depending on the country, including landfilling and recycling (e.g., subbase course material, aggregate for concrete, Fe-source for cement manufacturing). However, landfilling has several drawbacks, including environmental pollution due to leaching, high landfill costs, and wastage of iron resources. To improve Fe recovery in the clinker, we pulverized it into optimal -sized particles and employed specific gravity and magnetic force selection methods to isolate this metal. A carbon-based reducing agent and a binding material were added to the separated coarse powder (>10㎛) to prepare briquette clinker. A small amount (1-3%w/w) of the briquette clinker was charged with the scrap in an electric arc furnace to evaluate its feasibility as an additives (carbonaceous material, heat-generating material, and Fe source).

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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