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Production of Medium-chain-length Poly (3-hydroxyalkanoates) by Pseudomonas sp. EML8 from Waste Frying Oil (Pseudomonas sp. EML8 균주를 이용한 폐식용류로부터 medium-chain-length poly(3-hydroxyalkanoates) 생합성)

  • Kim, Tae-Gyeong;Kim, Jong-Sik;Chung, Chung-Wook
    • Journal of Life Science
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    • v.31 no.1
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    • pp.90-99
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    • 2021
  • In this study, to reduce the production cost of poly(3-hydroxyalkanoates) (PHA), optimal cell growth and PHA biosynthesis conditions of the isolated strain Pseudomonas sp. EML8 were established using waste frying oil (WFO) as the cheap carbon source. Gas chromatography (GC) and GC mass spectrometry analysis of the medium-chain-length PHA (mcl-PHAWFO) obtained by Pseudomonas sp. EML8 of WFO indicated that it was composed of 7.28 mol% 3-hydrxoyhexanoate, 39.04 mol% 3-hydroxyoctanoate, 37.11 mol% 3-hydroxydecanoate, and 16.58 mol% 3-hydroxvdodecanoate monomers. When Pseudomonas sp. EML8 were culture in flask, the maximum dry cell weight (DCW) and the mcl-PHAWFO yield (g/l) were showed under WFO (20 g/l), (NH4)2SO4 (0.5 g/l), pH 7, and 25℃ culture conditions. Based on this, the highest DCW, mcl-PHAWFO content, and mcl-PHAWFO yield from 3-l-jar fermentation was obtained after 48 hr. Similar results were obtained using 20 g/l of fresh frying oil (FFO) as a control carbon source. In this case, the DCW, the mcl-PHAFFO content, and the mcl-PHAFFO yields were 2.7 g/l, 62 wt%, and 1.6 g/l, respectively. Gel permeation chromatography analysis confirmed the average molecular weight of the mcl-PHAWFO and mcl-PHAFFO to be between 165-175 kDa. Thermogravimetric analysis showed decomposition temperature values of 260℃ and 274.7℃ for mcl-PHAWFO and mcl-PHAFFO, respectively. In conclusion, Pseudomonas sp. EML8 and WFO could be suggested as a new candidate and substrate for the industrial production of PHA.

Nanoscale Pattern Formation of Li2CO3 for Lithium-Ion Battery Anode Material by Pattern Transfer Printing (패턴전사 프린팅을 활용한 리튬이온 배터리 양극 기초소재 Li2CO3의 나노스케일 패턴화 방법)

  • Kang, Young Lim;Park, Tae Wan;Park, Eun-Soo;Lee, Junghoon;Wang, Jei-Pil;Park, Woon Ik
    • Journal of the Microelectronics and Packaging Society
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    • v.27 no.4
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    • pp.83-89
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    • 2020
  • For the past few decades, as part of efforts to protect the environment where fossil fuels, which have been a key energy resource for mankind, are becoming increasingly depleted and pollution due to industrial development, ecofriendly secondary batteries, hydrogen generating energy devices, energy storage systems, and many other new energy technologies are being developed. Among them, the lithium-ion battery (LIB) is considered to be a next-generation energy device suitable for application as a large-capacity battery and capable of industrial application due to its high energy density and long lifespan. However, considering the growing battery market such as eco-friendly electric vehicles and drones, it is expected that a large amount of battery waste will spill out from some point due to the end of life. In order to prepare for this situation, development of a process for recovering lithium and various valuable metals from waste batteries is required, and at the same time, a plan to recycle them is socially required. In this study, we introduce a nanoscale pattern transfer printing (NTP) process of Li2CO3, a representative anode material for lithium ion batteries, one of the strategic materials for recycling waste batteries. First, Li2CO3 powder was formed by pressing in a vacuum, and a 3-inch sputter target for very pure Li2CO3 thin film deposition was successfully produced through high-temperature sintering. The target was mounted on a sputtering device, and a well-ordered Li2CO3 line pattern with a width of 250 nm was successfully obtained on the Si substrate using the NTP process. In addition, based on the nTP method, the periodic Li2CO3 line patterns were formed on the surfaces of metal, glass, flexible polymer substrates, and even curved goggles. These results are expected to be applied to the thin films of various functional materials used in battery devices in the future, and is also expected to be particularly helpful in improving the performance of lithium-ion battery devices on various substrates.

Development and Testing of a RIVPACS-type Model to Assess the Ecosystem Health in Korean Streams: A Preliminary Study (저서성 대형무척추동물을 이용한 RIVPACS 유형의 하천생태계 건강성 평가법 국내 하천 적용성)

  • Da-Yeong Lee;Dae-Seong Lee;Joong-Hyuk Min;Young-Seuk Park
    • Korean Journal of Ecology and Environment
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    • v.56 no.1
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    • pp.45-56
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    • 2023
  • In stream ecosystem assessment, RIVPACS, which makes a simple but clear evaluation based on macroinvertebrate community, is widely used. In this study, a preliminary study was conducted to develop a RIVPACS-type model suitable for Korean streams nationwide. Reference streams were classified into two types(upstream and downstream), and a prediction model for macroinvertebrates was developed based on each family. A model for upstream was divided into 7 (train): 3 (test), and that for downstream was made using a leave-one-out method. Variables for the models were selected by non-metric multidimensional scaling, and seven variables were chosen, including elevation, slope, annual average temperature, stream width, forest ratio in land use, riffle ratio in hydrological characteristics, and boulder ratio in substrate composition. Stream order classified 3,224 sites as upstream and downstream, and community compositions of sites were predicted. The prediction was conducted for 30 macroinvertebrate families. Expected (E) and observed fauna (O) were compared using an ASPT biotic index, which is computed by dividing the BMWPK score into the number of families in a community. EQR values (i.e. O/E) for ASPT were used to assess stream condition. Lastly, we compared EQR to BMI, an index that is commonly used in the assessment. In the results, the average observed ASPT was 4.82 (±2.04 SD) and the expected one was 6.30 (±0.79 SD), and the expected ASPT was higher than the observed one. In the comparison between EQR and BMI index, EQR generally showed a higher value than the BMI index.

Selection and appropriate culture conditions of antagonistic bacterium Bacillus altitudinis HC7 against button mushroom cobweb disease caused by Cladobotryum mycophilum (양송이버섯 솜털곰팡이병균(Cladobotryum mycophilum)에 대한 길항미생물 Bacillus altitudinis HC7의 선발 및 적정 배양조건)

  • Chan-Jung Lee;Hye-Sung Park;Seong-Yeon Jo;Gi-Hong An;Ja-Yun Kim;Kang-Hyo Lee
    • Journal of Mushroom
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    • v.22 no.2
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    • pp.60-66
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    • 2024
  • This study was conducted to selection and investigate appropriate conditions for mass production of antagonistic microbes to control cobweb disease caused by Cladobotryum mycophilum. A grampositive bacterium was isolated from spent substrate of Agaricus bisporus and showed significant antagonistic activity against Cladobotryum mycophilum. The bacterium was identified as Bacillus altitudinis. based on the cultural, biochemical and physiological characteristics, and 16S rRNA sequence. The isolate is saprophytic, but not parasitic nor pathogenic to cultivated mushroom whereas it showed strong inhibitory effects against C. mycophilum cells in vitro. The control efficacy of B. altitudinis HC7 against cobweb disease of C. mycophilum was up to 78.2% on Agaricus bisporus. The suppressive bacterium may be useful for the development of biocontrol system. To define the appropriate conditions for the mass production of the Bacillus altitudinis HC7, we have investigated appropriate culture conditions and effects of various nutrient source on the bacterial growth. The appropriate initial pH and temperature were determined as pH 6.0 and 30℃, respectively. The appropriate concentration of medium elements for the growth of pathogen inhibitor bacterium(Bacillus altitudinis HC7) was determined as follows: 3.0% soluble startch, 10% soytone, 1.0% (NH4)2HPO4, 1.0 mmol KCl, and 0.5% L-asparagine.

Assessment of Methane Production Rate Based on Factors of Contaminated Sediments (오염퇴적물의 주요 영향인자에 따른 메탄발생 생성률 평가)

  • Dong Hyun Kim;Hyung Jun Park;Young Jun Bang;Seung Oh Lee
    • Journal of Korean Society of Disaster and Security
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    • v.16 no.4
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    • pp.45-59
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    • 2023
  • The global focus on mitigating climate change has traditionally centered on carbon dioxide, but recent attention has shifted towards methane as a crucial factor in climate change adaptation. Natural settings, particularly aquatic environments such as wetlands, reservoirs, and lakes, play a significant role as sources of greenhouse gases. The accumulation of organic contaminants on the lake and reservoir beds can lead to the microbial decomposition of sedimentary material, generating greenhouse gases, notably methane, under anaerobic conditions. The escalation of methane emissions in freshwater is attributed to the growing impact of non-point sources, alterations in water bodies for diverse purposes, and the introduction of structures such as river crossings that disrupt natural flow patterns. Furthermore, the effects of climate change, including rising water temperatures and ensuing hydrological and water quality challenges, contribute to an acceleration in methane emissions into the atmosphere. Methane emissions occur through various pathways, with ebullition fluxes-where methane bubbles are formed and released from bed sediments-recognized as a major mechanism. This study employs Biochemical Methane Potential (BMP) tests to analyze and quantify the factors influencing methane gas emissions. Methane production rates are measured under diverse conditions, including temperature, substrate type (glucose), shear velocity, and sediment properties. Additionally, numerical simulations are conducted to analyze the relationship between fluid shear stress on the sand bed and methane ebullition rates. The findings reveal that biochemical factors significantly influence methane production, whereas shear velocity primarily affects methane ebullition. Sediment properties are identified as influential factors impacting both methane production and ebullition. Overall, this study establishes empirical relationships between bubble dynamics, the Weber number, and methane emissions, presenting a formula to estimate methane ebullition flux. Future research, incorporating specific conditions such as water depth, effective shear stress beneath the sediment's tensile strength, and organic matter, is expected to contribute to the development of biogeochemical and hydro-environmental impact assessment methods suitable for in-situ applications.

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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