• Title/Summary/Keyword: Purification plant

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Purification and Characterization of a Serine Protease (CPM-2) with Fibrinolytic Activity from the Dung Beetles

  • Ahn, Mi-Young;Hahn, Bum-Soo;Ryu, Kang-Sun;Hwang, Jae-Sam;Kim, Yeong-Shik
    • Archives of Pharmacal Research
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    • v.28 no.7
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    • pp.816-822
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    • 2005
  • Catharsius protease-2 (CPM-2) was isolated from the body of dung beetles, Catharsius molossus, using a three step purification process (ammonium sulfate fractionation, gel filtration on Bio-Gel P-60, and affinity chromatography on DEAE Affi-Gel blue). The purified CPM-2, having a molecular weight of 24 kDa, was assessed homogeneously by SDS-polyacrylamide gel electrophoresis. The N-terminal amino acid sequence of CPM-2 was composed of X Val Gin Asp Phe Val Glu Glu lie Leu. CPM-2 was inactivated by $Cu^{2+}\;and\;Zn^{2+}$ and strongly inhibited by typical serine proteinase inhibitors such as TLCK, soybean trypsin inhibitor, aprotinin, benzamidine, and ${\alpha}_1$-antitrypsin. However, EDTA, EGTA, cysteine, $\beta$-mercaptoethanol, E64, and elastatinal had little effect on enzyme activity. In addition, antiplasmin and antithrombin III were not sensitive to CPM-2. Based on the results of a fibrinolytic activity test, CPM-2 readily cleaved $A{\alpha}-$ and $B{\beta}$-chains of fibrinogen and fibrin, and y-chain of fibrinogen more slowly. The nonspecific action of the enzyme resulted in extensive hydrolysis, releasing a variety of fibrinopeptides of fibrinogen and fibrin. Polyclonal antibodies of CPM-2 were reactive to the native form of antigen. The ELISA was applied to detect quantities, in nanograms, of the antigen in CPM-2 protein.

Effect of Sulfur Powder and Citric Acid on Arsenic Phytoremediation Using Pteris multifida in Forest Soil (봉의 꼬리를 이용한 수림지 토양의 비소정화에 미치는 유황분말과 구연산의 영향)

  • Kwon, Hyuk Joon;Cho, Ju Sung;Lee, Cheol Hee
    • Journal of the Korean Society of Environmental Restoration Technology
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    • v.17 no.1
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    • pp.1-12
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    • 2014
  • This study was carried out not only to identify the optimum concentrations of sulfur powder and citric acid treated for improving arsenic absorption of Pteris multifuda known as hyperaccumulator of arsenic, but also to develop arsenic purification model in the forest soil. After applying sulfur powder (0, 30, 45, $60g{\cdot}m^{-2}$) and citric acid (0, 200, 400, $800g{\cdot}m^{-2}$) in the forest soil contaminated with heavy metals, P. multifuda was planted and cultivated for 16 weeks. And then the growth and arsenic contents of plants were analyzed. In the result of research, the growth of P. mulifuda, except plant width, cultivated in soils treated with sulfur powder and citric acid was relatively lower than control. The accumulated amount of arsenic in aerial parts of P. multifuda ($1822.2mg{\cdot}kg^{-1}$) cultivated in soils treated with $200g{\cdot}m^{-2}$ citric acid was improved 62.5% against the control. And the accumulated amount of arsenic per 1 $m^2$ ($20.1mg{\cdot}m^{-2}$) was the greatest in $200g{\cdot}m^{-2}$ citric acid treatment. Translocation rate (TR) was higher in all acid treatment compare to control, and was the best in $200g{\cdot}m^{-2}$ citric acid treatment (0.95) especially. It showed that the arsenic absorbed in underground parts was transferred fast to aerial parts. Therefore, $200g{\cdot}m^{-2}$ citric acid treatment in the soil is recommended for arsenic purification using P. multifuda.

A Study of Filtralite Media Applicability for Development F/A Process of Membrane Filtration Pre-treatment Process in the Water Purification Plant (정수장에서 막여과 전처리용 F/A 공정 개발을 위한 Filtralite 여재의 적용성 연구)

  • Kim, JUN-Hyun;Jun, Yong-sung;Kwak, Young-ju;Jang, Jung Woo
    • Membrane Journal
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    • v.25 no.6
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    • pp.503-514
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    • 2015
  • In this study, water purification system without coagulant was consisted of F/A and membrane to produce purified water which only uses physical treatment without coagulant. Because the use of coagulant has a possibility remaining of hazardous aluminum with our health. Especially, the Filtralite was reviewed the possibility to remove turbidity and organic material. It was found that the turbidity removal rate of Filtralite was 83~84%. It show that Filtralite has similar efficiency to sand-filter. But Filtralite has higher 50% removal rate of organic material than sand-filter due to well-developed pore on the surface of it. So, Filtralite could be used to substitute the sand-filter for the F/A process due to higher removal rate. And also coupled with activated carbon in F/A process, TMP was increased by TOC value. To prevent increasing TMP, media that has outstanding organic adsorption ability should be used.

An Assessment on Efficiency of MBAS Removal in Urban Stream Maintenance Water by Using Sand Filtration (모래여과를 이용한 도시하천유지용수의 MBAS 제거 효율 평가)

  • Kim, hong bae;Ahn, kyung soo
    • Journal of Wetlands Research
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    • v.8 no.2
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    • pp.45-51
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    • 2006
  • Biological enhanced treatment and send filtration are established being operated to remove nutrients and MBAS(Methylene Blue Activate Substance) in the most of Waste Water Treatment Plant(WWTP) in Korea. However, untreated synthetic detergents and nutrients which directly run into the water system present an unpleasant view because of the foam, taste and odor generating filamentous periphytic algae and interrupting self-purification in the stream. Therefore, this research was enforced to know the MBAS removal efficiency of the sand filtration about G WWTP which reuses effluent as urban stream management water. As a result, the maximum removal efficiency using sand filtration was 63% after 24 hours and particularly 30% after 2 or 4 hours which turned out to be not that effective. In conclusion, It is recognized that other methods of MBAS removal and a research will be needed which reuse effluent as urban stream management water from now on. Because the MBAS removal with sand filtration is insufficient with economical efficiency from the fact that it needs long hours for a sand filtration treatment and the removal efficiency was almost below the expectation.

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Purification of ginseng rare sapogenins 25-OH-PPT and its hypoglycemic, antiinflammatory and lipid-lowering mechanisms

  • Xu, Jing;Liu, Hairong;Su, Guangyue;Ding, Meng;Wang, Wei;Lu, Jincai;Bi, Xiuli;Zhao, Yuqing
    • Journal of Ginseng Research
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    • v.45 no.1
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    • pp.86-97
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    • 2021
  • Background: Panax ginseng Meyer has been used as a nourishing edible herb in East Asia for thousands of years. 25-OH-PPT was first discovered as a natural rare triterpenoid saponin in ginseng stems and leaves by our group. Research found that it showed strong inhibitory effects on α-glucosidase and protein tyrosine phosphatase 1B, and protected cardiocytes (H9c2) through PI3K/Akt pathway. Methods: In the research, in order to optimize the 25-OH-PPT enrichment process, optimal macroporous resins and optimal purification conditions were studied. Meanwhile, the hypoglycemic effect and mechanism of 25-OH-PPT were evaluated by using STZ to establish insulin-dependent diabetic mice and the spontaneous type 2 diabetes DB/DB mice. Results and Conclusion: Research found that 25-OH-PPT can reduce blood glucose and enhance glucose tolerance in STZ model mice. It increases insulin sensitivity by upregulating GLUT4 and AMPK in skeletal muscle, and activating insulin signaling pathways. In DB/DB mice, 25-OH-PPT achieves hypoglycemic effects mainly by activating the insulin signaling pathway. Meanwhile, through the influence of liver inflammatory factors and lipids in serum, it can be seen that 25-OH-PPT has obvious anti-inflammatory and lipid-lowering effects. These results provide new insights into the study of ginseng as a functional food.

Genomics and LC-MS Reveal Diverse Active Secondary Metabolites in Bacillus amyloliquefaciens WS-8

  • Liu, Hongwei;Wang, Yana;Yang, Qingxia;Zhao, Wenya;Cui, Liting;Wang, Buqing;Zhang, Liping;Cheng, Huicai;Song, Shuishan;Zhang, Liping
    • Journal of Microbiology and Biotechnology
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    • v.30 no.3
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    • pp.417-426
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    • 2020
  • Bacillus amyloliquefaciens is an important plant disease-preventing and growth-promoting microorganism. B. amyloliquefaciens WS-8 can stimulate plant growth and has strong antifungal properties. In this study, we sequenced the complete genome of B. amyloliquefaciens WS-8 by Pacific Biosciences RSII (PacBio) Single Molecule Real-Time (SMRT) sequencing. The genome consists of one chromosome (3,929,787 bp) and no additional plasmids. The main bacteriostatic substances were determined by genome, transcriptome, and mass spectrometry data. We thereby laid a theoretical foundation for the utilization of the strain. By genomic analysis, we identified 19 putative biosynthetic gene clusters for secondary metabolites, most of which are potentially involved in the biosynthesis of numerous bioactive metabolites, including difficidin, fengycin, and surfactin. Furthermore, a potential class II lanthipeptide biosynthetic gene cluster and genes that are involved in auxin biosynthesis were found. Through the analysis of transcriptome data, we found that the key bacteriostatic genes, as predicted in the genome, exhibited different levels of mRNA expression. Through metabolite isolation, purification, and exposure experiments, we found that a variety of metabolites of WS-8 exert an inhibitory effect on the necrotrophic fungus Botrytis cinerea, which causes gray mold; by mass spectrometry, we found that the main substances are mainly iturins and fengycins. Therefore, this strain has the potential to be utilized as an antifungal agent in agriculture.

Isolation and In vitro and In vivo Antifungal Activity of Phenylacetic acid Produced by Micromonospora aurantiaca Strain JK-1

  • Kim, Hyo-Jin;Hwang, In-Sun;Kim, Beom-Seok;Hwang, Byung-Kook
    • The Plant Pathology Journal
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    • v.22 no.1
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    • pp.75-89
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    • 2006
  • The actinomycete strain JK-1 that showed strong inhibitory activity against some plant pathogenic fungi and oomycetes was isolated from Jung-bal Mountain in Ko-yang, Korea. The strain JK-1 produced spores singly borne on sporophores and the spores were spherical and 0.9-1.2 11m in diameter. The cell wall of the strain JK-1 contained meso-diaminopimelic acid. The actinomycete strain JK-1 was identified as the genus Micromonospora based on the morphological, physiological, biochemical and chemotaxonomic characteristics. From the 168 rDNA analysis, the strain JK-1 was assigned to M aurantiaca. The antibiotic MA-1 was purified from the culture broth of M aurantiaca JK-1 using various purification procedures, such as Diaion HP20 chromatography, C18 flash column chromatography, silica gel flash column chromatography and Sephadex LH-20 column chromatography. $^{1}H-$, $^{13}C-NMR$ and EI mass spectral analysis of the antibiotic MA-1 revealed that the antibiotic MA-1 is identical to phenylacetic acid. Phenylacetic acid showed in vitro inhibitory effects against fungal and oomycete pathogens Alternaria mali, Botrytis cinerea, Magnaporthe grisea, Phytophthora capsici and yeast Saccharomyces cerevisiae at < 100 $\mug$ $ml^{-1}$. In addition, phenylacetic, acid completely inhibited the growth of Sclerotinia sclerotiorum, Bacillus subtilis, Candida albicans, Xanthomonas campestris pv. vesicatoria at < $\mug$ $ml^{-1}$. Phenylacetic acid strongly inhibited conidial germination and hyphal growth of M grisea and C. orbiculare. Phenylacetic acid showed significantly high levels of inhibitory' effect against rice blast and cucumber anthracnose diseases at 250 $\mug$ $ml^{-1}$. The control efficacies of phenylacetic acid against the two diseases were similar to those of commercial compounds tricyclazole, iprobenfos and chlorothalonil .n the greenhouse.

Freeze Tolerance Enhanced by Antifreeze Protein in Plant

  • Hwang, Cheol-Ho;Park, Hyun-Woo;Min, Sung-Ran;Liu, Jang-Ryol
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.4
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    • pp.339-343
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    • 2000
  • When plants are exposed to subfreezing temperatures ice crystals are forming within extracelluar space in leaves. The growth of ice crystal is closely related to the degree of freezing injury. It was shown that an antifreeze protein binds to an ice nucleator through hydrogen bonds to prevent growth of ice crystal and also reduces freezing damage. The antifreeze proteins in plants are similar to PR proteins but only the PR proteins induced upon cold acclimation were shown to have dual functions in antifreezing as well as antifungal activities. Three of the genes encoded for CLP, GLP, and TLP were isolated from barley and Kentucky bluegrass based on amino acid sequence revealed after purification and low temperature-inducibility as shown in analysis of the protein. The deduced amino acid of the genes cloned showed a signal for secretion into extracellular space where the antifreezing activity sup-posed to work. The western analysis using the antisera raised against the antifreeze proteins showed a positive correlation between the amount of the protein and the level of freeze tolerance among different cultivars of barely. Besides it was revealed that TLP is responsible for a freeze tolerance induced by a treatment of trinexapac ethyl in Kentucky bluegrass. Analysis of an overwintering wild rice, Oryza rufipogon also showed that an acquisition of freeze tolerance relied on accumulation of the protein similar to CLP. The more direct evidence for the role of CLP in freeze tolerance was made with the analysis of the transgenic tobacco showing extracellular accumulation of CLP and enhanced freeze tolerance measured by amount of ion leakage and rate of photosynthetic electron transport upon freezing. These antifreeze proteins genes will be good candidates for transformation into crops such as lettuce and strawberry to develop into the new crops capable of freeze-storage and such as rose and grape to enhance a freeze tolerance for a safe survival during winter.

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Purification and Phytotoxicity of Apicidins Produced by the Fusarium semitectum KCTC16676

  • Jin, Jianming;Baek, Seung-Ryel;Lee, Kyung-Rim;Lee, Jungkwan;Yun, Sung-Hwan;Kang, Seog-Chan;Lee, Yin-Won
    • The Plant Pathology Journal
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    • v.24 no.4
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    • pp.417-422
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    • 2008
  • Apicidin is a cyclic tetrapeptide produced by some Fusarium species and is known to inhibit Apicomplexan histone deacetylase. The goals of this study were to determine species identity of Fusarium isolate KCTC16676, an apicidin producer, to improve a method for apicidin extraction, and to test phytotoxicity of apicidin and its analogs. We compared sequences of the translation elongation factor 1-alpha (TEF) gene in KCTC16676 with those from isolates representing diverse Fusarium species, which showed that KCTC16676 belongs to the F. semitectum-F. equiseti species complex. To enhance apicidin production, after culturing isolate KCTC16676 on a wheat medium for 3 weeks at $25^{\circ}C$, the culture was extracted with chloroform. Apicidins were purified through a reverse phase $C_{18}$ silica gel column, resulting in 5 g of apicidin, 200 mg of apicidin A, and 300 mg of apicidin $D_2$ from 4 kg of wheat cultures; this represents a significant yield improvement from a previous method, offers more materials to study the modes of its action, and facilitates the elucidation of the apicidin biosynthesis pathway. Apicidin and apicidin $D_2$ showed phytotoxicity on both seedlings and 2-week-old plants of diverse species, and weeds were more sensitive to apicidins than vegetables

Treatment Efficiency of a Subsurface-Flow Wetland System Constructed on Floodplain (고수부지를 이용한 여과습지의 수질정화 초기처리)

  • Yang, Hongmo
    • Journal of the Korean Society of Environmental Restoration Technology
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    • v.4 no.4
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    • pp.56-63
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    • 2001
  • This paper presents treatment efficiency and plant growth of a subsurface-flow constructed wetland system (23 m in length, 6.5 m in width, 0.65 m in depth) over one year after its establishment on floodplain of a stream in June 2000. An upper layer of 10 cm in depth was filled with course sand and the main biological layer of 50 cm depth with crushed stone with 8 - 15 mm in diameter. The system was planted with common reeds (Phragmites australis) grown on pots. Effluent discharged from a secondary-level treatment plant was funneled into it. Reed stems emerging in April 2001 grew up to 145.9cm until July 2001. The number of reed stems in July 2001 increased by about 11 times compared with that just after planting. The system was inundated seven times by storms over the monitoring period. Reeds were slightly bent after flooding, however they returned to almost upright standing in a couple of weeks. Small portion of inside slope of berm was eroded and the system surface had a sedimentation of 2 - 3 mm in depth. The average removal rates for SS, $BOD_5$, T-N and T-P was 73%, 70%, 53%, and 72%, respectively. The purification efficiencies for SS and $BOD_5$ were fairly good. The reduction rates for T-N was relatively low for the period of late fall through winter until early spring due to lower water temperature which retarded microbial nitrification and denitrification mechanisms. Reduction in the concentration of T-P during fall and winter was relatively higher than that during spring. Leach of phosphorous from plant litters lying on system surface and slight resuspension of precipitated phosphorous in substrates resulted in lower reduction for T-P in spring.

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