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Enzymatic Hydrolysis of Rice Straw, a Lignocellulosic Biomass, by Extracellular Enzymes from Fomitopsis palustris (Fomitopsis palustris의 균체 외 효소에 의한 볏짚 당화에 관한 연구)

  • Kim, Yoon-Hee;Cho, Moon-Jung;Shin, Keum;Kim, Tae-Jong;Kim, Nam-Hun;Kim, Yeong-Suk
    • Journal of the Korean Wood Science and Technology
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    • v.38 no.3
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    • pp.262-273
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    • 2010
  • In the enzymatic hydrolysis of rice straw and wood meals using extra-cellular enzymes from Fomitopsis palustris, key factors which enhanced the sugar conversion yield were investigated in this work, such as enzyme production and enzyme reaction conditions, surfactant effects, and the surface structure of substrates. F. palustris cultured with softwood mixture produced 12.0 U/$m{\ell}$ for endo-${\beta}$-1,4-gulcanase (EG), 116.68 U/$m{\ell}$ for ${\beta}$-glucosidase (BGL), 18.82 U/$m{\ell}$ for cellobiohydrolase (CBH), and 13.33 U/$m{\ell}$ for ${\beta}$-xylosidase (BXL). These levels of BGL, CBH, and BXL activities were two to four folds more than enzyme activities of F. palustris cultured with rice straw. The optimum reaction conditions of cellulase-RS which produced by F. palustris with rice straw and cellulase-SW which produced by F. palustris with softwood mixture were pH 5.0 at $45^{\circ}C$ and pH 5.0 at $50^{\circ}C$, respectively. The sugar conversion yield of cellulase-SW had the highest value of $40.6{\pm}0.6%$ within 72 h when rice straw was used as substrate. By adding 0.1% Tween 20 (w/w-substrate), the sugar conversion yield of rice straw was increased to 44%, which was about four fifths sugar conversion yield of commercial enzyme, Celluclast 1.5L (Novozyme A/S). A low crystallinity and an intensive fibril surface observed by the scanning electron microscope may explain the high sugar conversion yield of rice straw.

Effects of Initial Shoot, Root Length, and Acclimating Substrates on Survival Rate of Plantlets Regenerated from Somatic Embryos of Larix kaempferi (일본잎갈나무 체세포배 유래 식물체의 초기 신초와 뿌리 길이, 순화용 기질이 생존율에 미치는 영향)

  • Lee, Na Nyum;Yun, A Young;Kim, Ji Ah;Kim, Tae Dong;Kim, Yong Wook;Han, Sim Hee
    • Journal of Korean Society of Forest Science
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    • v.109 no.4
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    • pp.413-420
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    • 2020
  • We analyzed the growth characteristics of each cell line and acclimating substrate of Larix kaempferi plantlets regenerated from somatic embryos, with the goal of increasing the survival rate during the acclimation phase. Somatic embryos from three embryogenic cell lines (L14-66, L16-18, and L17-B4) were used, and the acclimating substrates were commercial soils for Larix species (Larix-Soil) and horticultural corps (Hort-Soil), Elle-pot, and Peat-plug. The average initial shoot and root length was shortest in L14-66 and longest in L17-B4. The average survival rate by cell line was highest (87.0%) in L17-B4 and lowest (64.3%) for L14-66. Survival rates by substrate were highest in Elle-pot (88.5%) and Peat-plug (88.9%). The initial shoot length of the L14-66 plantlets was highly correlated with survival rates in the Larix-Soil (r = 0.852), Hort-Soil (r = 0.692), and Elle-pot (r = 0.867) substrates, but not in Peat-plug with high total nitrogen content. The initial shoot length of the L17-B4 plantlets was not correlated with the survival rate in any of the substrates. The initial root length of the L14-66 plantlets was highly related to survival rates in the Larix-Soil (r = 0.986), Elle-pot (r = 0.846), and Peat-plug (r = 0.802) substrates, and the survival rate of the plantlets was higher as the initial root length was longer. The initial root length of the L17-B4 plantlets was related to survival rate only in the Larix-Soil (r = 0.896) and Elle-pot (r = 0.696) substrates. In conclusion, to increase the survival rate of plantlets, root length should be considered over shoot length, and it is recommended to use substrates with high nitrogen content, such as Peat-plug, or to add fertilizer, during the acclimating process. In addition, in order to increase the survival rate, lines with high initial growth should be developed.

Characterization of an Antarctic alkaline protease, a cold-active enzyme for laundry detergents (세탁세제 첨가용 효소 개발을 위한 남극 해양세균 유래 저온성 단백질분해효소의 특성 연구)

  • Park, Ha Ju;Han, Se Jong;Yim, Joung Han;Kim, Dockyu
    • Korean Journal of Microbiology
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    • v.54 no.1
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    • pp.60-68
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    • 2018
  • A cold-active and alkaline serine protease (Pro21717) was partially purified from the Antarctic marine bacterium Pseudoalteromonas arctica PAMC 21717. On a zymogram gel containing skim milk, Pro21717 produced two distinct clear-zones of approximately 37 kDa (low intensity) and 74 kDa (high intensity). These were found to have identical N-terminal sequences, suggesting they arose from an identical precursor and that the 37 kDa protease might homodimerize to the more active 74 kDa form of the protein. Pro21717 displayed proteolytic activity at $0-40^{\circ}C$ (optimal temperature of $40^{\circ}C$) and maintained this activity at pH 5.0-10.0 (optimal pH of 9.0). Notably, relative activities of 30% and 45% were observed at $0^{\circ}C$ and $10^{\circ}C$, respectively, in comparison to the 100% activity observed at $40^{\circ}C$, and this enzyme showed a broad substrate range against synthetic peptides with a preference for proline in the cleavage reaction. Pro21717 activity was enhanced by $Cu^{2+}$ and remained stable in the presence of detergent surfactants (linear alkylbenzene sulfonate and sodium dodecyl sulfate) and other chemical components ($Na_2SO_4$ and metal ions, such as $Ba^{2+}$, $Mg^{2+}$, $Ca^{2+}$, $Zn^{2+}$, $Fe^{2+}$, $K^+$, and $Na^{2+}$), which are often included in commercial detergent formulations. These data indicate that the psychrophilic Pro21717 has properties comparable to the well-characterized mesophilic subtilisin Carlsberg, which is commercially produced by Novozymes as the trademark Alcalase. Thus it has the potential to be used as a new additive enzyme in laundry detergents that must work well in cold tap water below $15^{\circ}C$.

Effects of Detoxified Sulfur as a Feed Supplement on in Vitro Rumen Fermentation and Methane Mitigation (제독 유황의 반추위 발효성상 및 메탄 저감 효과 연구)

  • Kim, Seon-Ho;Islam, Mahfuzul;Biswas, Ashraf Ali;Cho, Kwang-Keun;Lee, Sang-Suk
    • Journal of Life Science
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    • v.30 no.9
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    • pp.743-748
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    • 2020
  • Sulfate is a reductant that competes for electrons and may lower CH4 production in the rumen. This study was designed to evaluate the beneficial effect of detoxified sulfur powder supplementation on in vitro rumen fermentation and methane mitigation. A ruminally cannulated Holstein Friesian cow was used as a rumen fluid source, and commercial pelleted concentrate was used as a substrate at 1 g dry matter. Treatments included the addition of detoxified sulfur powder at the rate of 0% (Control), 0.2% (T1), 0.4% (T2), 0.6% (T3), 0.8% (T4), and 1.0% (T5) as dry matter (DM) basis. The pH, total gas (TG), methane (CH4) production, DM digestibility, organic matter (OM) digestibility, and volatile fatty acids (VFA) production were analyzed after 12 hr of incubation. The results showed that CH4 production was significantly lowest in T1 (13.78 ml) but highest in the control (20.16 ml). Insignificantly higher total VFA was observed in control and T1 (64.99 and 64.28 mM, respectively) compared to other treatments after 12 hr of incubation. After 12 hr of incubation, the significantly lowest acetate:propionate was observed in T1 (1.90) while the highest was observed in T4 (2.44). However, no significant differences were recorded for pH, TG, DM digestibility, OM digestibility, acetate, propionate, and butyrate between the control and T1. Total number of bacterial DNA copies was significantly lower in the treatment group than the control. Therefore, it can be concluded from this study that detoxified sulfur at 0.2% inclusion level is optimal for production performance and ruminal CH4 mitigation.

Selection and Cultural Characteristics of Whole Chicken Feather-Degrading Bacterium, Bacillus sp. SMMJ-2 (Whole Chicken Feather-Degrading Keratinolytic Protease 생산균주의 분리 및 특성)

  • Park Sung-Min;Jung Hyuck-Jun;Yu Tae-Shick
    • Microbiology and Biotechnology Letters
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    • v.34 no.1
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    • pp.7-14
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    • 2006
  • Feather, generated in large quantities as a byproduct of commercial poultry processing, is almost pure keratin, which is not easily degradable by common professes. Four strains, SMMJ-2, FL-3, NO-4 and RM-12 were isolated from soil for production of extracellular keratinolytic protease. They were identified as Bacillus sp. based on their morphological and physiological characteristics. They shown high protease activity on 5.0% skim milk agar medium and produced a substrate like mucoid on keratin agar medium. Bacillus sp. SMMJ-2 had a faster production time for producing keratinolytic protease than other strains. This strain did not completely degrade whole chicken feather for five days in basal medium but completely degraded whole chicken feather when supplied with nitrogen source for 40hours in keratinolytic producing medium ($0.7%\;K_{2}HPO_{4},\;0.2%\;KH_{2}PO_{4},\;0.1%$ fructose, 1.2% whole chicken feather, $0.01%\;Na_{2}CO_3$, pH 7.0). When supplied with chicken feather as nitrogen source, keratinolytic protease activity was 89 units/ml/min. When soybean meal was used as nitrogen source, the keratinolytic protease production reached a maximum of 106 units/ml/min after 48 hours under $30^{\circ}C$, 180 agitation. To isolate the keratinolytic protease, the culture filtrate was precipitated with $(NH_4)_{2}SO_4$ and acetone. The recovery rate of keratinolytic protease was about 96% after treatment with 50% acetone. The enzyme was stable in the range of $30{\sim}50^{\circ}C$ and pH $6.0{\sim}12.0$.

Effect of Highly Concentrated Turbid Water on the Water Quality and Periphytic Diatom Community in Artificial Channel (인공수로에서 고농도 탁수가 수질 및 부착 규조류 군집에 미치는 영향)

  • Yoon, Sung-Ae;You, Kyung-A;Park, Ji-Hyoung;Kim, Baik-Ho;Hwang, Soon-Jin
    • Korean Journal of Ecology and Environment
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    • v.44 no.1
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    • pp.75-84
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    • 2011
  • We examined the effect of the turbid water on the periphytic diatom community in an artificial stream system. The artificial stream was constructed with transparent acryl and composed of four channels. Each channel ($20\;cm{\times}200\;cm{\times}40\;cm$) was supplied continuously with eutrophic lake water. In order to the freely colonize and grow diatoms, artificial substrate was installed with commercial slide glass soaked in 1% agar. Prior to introducing turbid water, the artificial stream was operated with lake water for 6 days to permit the propagation of diatom community on the substrates. The turbid water prepared with sediment sieved with ${\varphi}$ $64\;{\mu}m$ at $2\;g\;L^{-1}$ (final concentration, 300 NTU) was provided daily for 50 minute duration. The experiment was conducted for 7 days with manipulated experimental condition of light ($50{\sim}80\;{\mu}mol\;m^{-2}s^{-1}$, light:dark=24:0), temperature ($10{\pm}1^{\circ}C$), and flow rate ($0.31\;cm\;s^{-1}$). Sampling and analysis were conducted daily for water quality and diatom. Turbidity of the water varied 162.2~173.2 NTU during the experiment. After introduction of turbid water, DO, pH and TN were decreased, while SS and TP increased significantly. A total of 14 genera and 47 species of diatoms was observed on the artificial substrates during the experimental period. Of these, Navicula appeared to be a most dominant genus with 10 species, followed by Cymbella (6 species), Fragilaria (6 species) and Gomphonema (5 species). Achnanthes minutissima was the most dominant species (>70% of total frequency) in both control and treatment experiments. Increase in diatom abundance lasted for three days since turbid water introduction, after that they gradually decreased by the termination of the experiment. These results suggest that frequent supply of highly-concentrated turbid water significantly decreases the periphytic diatom community, and retard the recovery of the stable food-web within the stream.

Effect of SO2 on NOx Removal Performance in Low Temperature Region over V2O5-Sb2O3/TiO2 SCR Catalyst Washcoated on the Metal Foam (저온영역에서 메탈폼에 코팅된 V2O5-Sb2O3/TiO2 SCR 촉매의 NOx 저감성능에 미치는 SO2 영향에 관한 연구)

  • Na, Woo-Jin;Park, Young-Jin;Bang, Hyun-Seok;Bang, Jong-Seong;Park, Hea-Kyung
    • Clean Technology
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    • v.22 no.2
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    • pp.132-138
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    • 2016
  • The emission of SO2 is inevitable in case of combustion of most fossil fuels except LNG in commercial power plant which has a bad effect on the durability of SCR catalyst. To develop a low temperature SCR catalyst which has a high NOx removal performance and excellent durability to SO2, V2O5/TiO2 catalysts were prepared by coating on the metal foam substrate with the impregnation amount of Sb2O3 as promotor. This study has evaluated the NOx removal performance and the durability to SO2 on a laboratory scale atmospheric reactor and analyzed the properties of the prepared catalysts by means of porosimeter, BET, SEM (scanning electron microscope), EDX (energy dispersive x-ray spectrometer), XPS (X-ray photoelectron spectroscopy). It was found that the surface area of catalyst increased with the impregnation amount of Sb2O3 and the NOx removal performance showed the highest value at the 2 wt% impregnation of Sb2O3. This results was considered to be due to the optimum active site on the catalyst surface. And also, Sb2O3 impregnated catalysts presented that NOx removal performance was maintained despite the exposure to SO2 for 5 hours. Therefore it was confirmed that metal foam SCR catalyst for low temperature could be manufactured with the optimum control of Sb2O3 impregnation according to the SO2 presence or not.

Evaluation of Fruit Yield and Quality of Netted Melon, Water and Nutrient Use Efficiency in a Closed Hydroponic System (순환식 수경재배 멜론의 수량과 품질, 관개수 및 양분 이용 효율성 평가)

  • Minju Shin;Seungri Yoon;Jin Hyun Kim;Ho Jeong Jeong;Sung Kyeom Kim
    • Journal of Bio-Environment Control
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    • v.32 no.4
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    • pp.492-500
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    • 2023
  • The spectrum of this study was research on the closed hydroponic cultivation of netted melons (Cucumis melo L.) using coir substrate, analyzing the impact of this cultivation method on melon yield, fruit quality, and the efficiency of water and nutrient usage. The experimental results showed that the average fruit weight of the melons grown in a closed system was 71.4 g higher than that of the open system, and the fruit width was on average 0.2 cm larger, showing a statistically significant difference. However, there was no difference in the average sugar content of the fruit flesh and height. Although there is no substantial commercial difference, it is conjectured that the change in the macronutrients ratio in the irrigation has played a role in the statistically significant increase in fruit weight, which is attributed to changes in the crops' nutrient uptake concentrations. This necessitates further research for a more comprehensive understanding. In terms of the productivity of irrigation required to produce the fruit, applying the closed system resulted in an increase of 7.6 kg/ton compared to the open system, saving 31.6% of water resources. Additionally, in terms of nutrients, cultivating in a closed system allowed for savings of approximately 59, 25, 55, 83, 76, and 87% of N, P, K, Ca, Mg, and S, respectively, throughout the entire cultivation period. As the drainage was reused, the ratios of NO3- and Ca2+ increased up to a maximum of 9.6 and 9.1%, respectively, while the ratios of other ions gradually decreased. In summary, these results suggest that closed hydroponic cultivation can effectively optimize the use of water and fertilizer while maintaining excellent fruit quality in melon cultivation.

Brief Introduction of Research Progresses in Control and Biocontrol of Clubroot Disease in China

  • He, Yueqiu;Wu, Yixin;He, Pengfei;Li, Xinyu
    • 한국균학회소식:학술대회논문집
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    • 2015.05a
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    • pp.45-46
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    • 2015
  • Clubroot disease of crucifers has occurred since 1957. It has spread to the whole China, especially in the southwest and nourtheast where it causes 30-80% loss in some fields. The disease has being expanded in the recent years as seeds are imported and the floating seedling system practices. For its effective control, the Ministry of Agriculture of China set up a program in 2010 and a research team led by Dr. Yueqiu HE, Yunnan Agricultural University. The team includes 20 main reseachers of 11 universities and 5 institutions. After 5 years, the team has made a lot of progresses in disease occurrence regulation, resources collection, resistance identification and breeding, biological agent exploration, formulation, chemicals evaluation, and control strategy. About 1200 collections of local and commercial crucifers were identified in the field and by artificiall inoculation in the laboratories, 10 resistant cultivars were breeded including 7 Chinese cabbages and 3 cabbages. More than 800 antagostic strains were isolated including bacteria, stretomyces and fungi. Around 100 chemicals were evaluated in the field and greenhouse based on its control effect, among them, 6 showed high control effect, especially fluazinam and cyazofamid could control about 80% the disease. However, fluzinam has negative effect on soil microbes. Clubroot disease could not be controlled by bioagents and chemicals once when the pathogen Plasmodiophora brassicae infected its hosts and set up the parasitic relationship. We found the earlier the pathogent infected its host, the severer the disease was. Therefore, early control was the most effective. For Chinese cabbage, all controlling measures should be taken in the early 30 days because the new infection could not cause severe symptom after 30 days of seeding. For example, a biocontrol agent, Bacillus subtilis Strain XF-1 could control the disease 70%-85% averagely when it mixed with seedling substrate and was drenching 3 times after transplanting, i.e. immediately, 7 days, 14 days. XF-1 has been deeply researched in control mechanisms, its genome, and development and application of biocontrol formulate. It could produce antagonistic protein, enzyme, antibiotics and IAA, which promoted rhizogenesis and growth. Its The genome was sequenced by Illumina/Solexa Genome Analyzer to assembled into 20 scaffolds then the gaps between scaffolds were filled by long fragment PCR amplification to obtain complet genmone with 4,061,186 bp in size. The whole genome was found to have 43.8% GC, 108 tandem repeats with an average of 2.65 copies and 84 transposons. The CDSs were predicted as 3,853 in which 112 CDSs were predicted to secondary metabolite biosynthesis, transport and catabolism. Among those, five NRPS/PKS giant gene clusters being responsible for the biosynthesis of polyketide (pksABCDEFHJLMNRS in size 72.9 kb), surfactin(srfABCD, 26.148 kb, bacilysin(bacABCDE 5.903 kb), bacillibactin(dhbABCEF, 11.774 kb) and fengycin(ppsABCDE, 37.799 kb) have high homolgous to fuction confirmed biosynthesis gene in other strain. Moreover, there are many of key regulatory genes for secondary metabolites from XF-1, such as comABPQKX Z, degQ, sfp, yczE, degU, ycxABCD and ywfG. were also predicted. Therefore, XF-1 has potential of biosynthesis for secondary metabolites surfactin, fengycin, bacillibactin, bacilysin and Bacillaene. Thirty two compounds were detected from cell extracts of XF-1 by MALDI-TOF-MS, including one Macrolactin (m/z 441.06), two fusaricidin (m/z 850.493 and 968.515), one circulocin (m/z 852.509), nine surfactin (m/z 1044.656~1102.652), five iturin (m/z 1096.631~1150.57) and forty fengycin (m/z 1449.79~1543.805). The top three compositions types (contening 56.67% of total extract) are surfactin, iturin and fengycin, in which the most abundant is the surfactin type composition 30.37% of total extract and in second place is the fengycin with 23.28% content with rich diversity of chemical structure, and the smallest one is the iturin with 3.02% content. Moreover, the same main compositions were detected in Bacillus sp.355 which is also a good effects biocontol bacterial for controlling the clubroot of crucifer. Wherefore those compounds surfactin, iturin and fengycin maybe the main active compositions of XF-1 against P. brassicae. Twenty one fengycin type compounds were evaluate by LC-ESI-MS/MS with antifungal activities, including fengycin A $C_{16{\sim}C19}$, fengycin B $C_{14{\sim}C17}$, fengycin C $C_{15{\sim}C18}$, fengycin D $C_{15{\sim}C18}$ and fengycin S $C_{15{\sim}C18}$. Furthermore, one novel compound was identified as Dehydroxyfengycin $C_{17}$ according its MS, 1D and 2D NMR spectral data, which molecular weight is 1488.8480 Da and formula $C_{75}H_{116}N_{12}O_{19}$. The fengycin type compounds (FTCPs $250{\mu}g/mL$) were used to treat the resting spores of P. brassicae ($10^7/mL$) by detecting leakage of the cytoplasm components and cell destruction. After 12 h treatment, the absorbencies at 260 nm (A260) and at 280 nm (A280) increased gradually to approaching the maximum of absorbance, accompanying the collapse of P. brassicae resting spores, and nearly no complete cells were observed at 24 h treatment. The results suggested that the cells could be lyzed by the FTCPs of XF-1, and the diversity of FTCPs was mainly attributed to a mechanism of clubroot disease biocontrol. In the five selected medium MOLP, PSA, LB, Landy and LD, the most suitable for growth of strain medium is MOLP, and the least for strains longevity is the Landy sucrose medium. However, the lipopeptide highest yield is in Landy sucrose medium. The lipopeptides in five medium were analyzed with HPLC, and the results showed that lipopeptides component were same, while their contents from B. subtilis XF-1 fermented in five medium were different. We found that it is the lipopeptides content but ingredients of XF-1 could be impacted by medium and lacking of nutrition seems promoting lipopeptides secretion from XF-1. The volatile components with inhibition fungal Cylindrocarpon spp. activity which were collect in sealed vesel were detected with metheds of HS-SPME-GC-MS in eight biocontrol Bacillus species and four positive mutant strains of XF-1 mutagenized with chemical mutagens, respectively. They have same main volatile components including pyrazine, aldehydes, oxazolidinone and sulfide which are composed of 91.62% in XF-1, in which, the most abundant is the pyrazine type composition with 47.03%, and in second place is the aldehydes with 23.84%, and the third place is oxazolidinone with 15.68%, and the smallest ones is the sulfide with 5.07%.

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