• Title/Summary/Keyword: Purification plant

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Cloning, Sequencing, and Expression of the Gene Encoding a Multidomain Endo-$\beta$-1,4-Xylanase from Paenibacillus curdlanolyticus B-6, and Characterization of the Recombinant Enzyme

  • Waeonukul, Rattiya;Pason, Patthra;Kyu, Khin Lay;Sakka, Kazuo;Kosug, Akihiko;Mori, Yutaka;Ratanakhanokchai, Khanok
    • Journal of Microbiology and Biotechnology
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    • v.19 no.3
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    • pp.277-285
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    • 2009
  • The nucleotide sequence of the Paenibacillus curdlanolyticus B-6 xyn10A gene, encoding a xylanase Xyn10A, consists of 3,828 nucleotides encoding a protein of 1,276 amino acids with a predicted molecular mass of 142,726 Da. Sequence analysis indicated that Xyn10A is a multidomain enzyme comprising nine domains in the following order: three family 22 carbohydrate-binding modules (CBMs), a family 10 catalytic domain of glycosyl hydrolases (xylanase), a family 9 CBM, a glycine-rich region, and three surface layer homology (SLH) domains. Xyn10A was purified from a recombinant Escherichia coli by a single step of affinity purification on cellulose. It could effectively hydrolyze agricultural wastes and pure insoluble xylans, especially low substituted insoluble xylan. The hydrolysis products were a series of short-chain xylooligosaccharides, indicating that the purified enzyme was an endo-$\beta$-1,4-xylanase. Xyn10A bound to various insoluble polysaccharides including Avicel, $\alpha$-cellulose, insoluble birchwood and oat spelt xylans, chitin, and starches, and the cell wall fragments of P. curdlanolyticus B-6, indicating that both the CBM and the SLH domains are fully functioning in the Xyn10A. Removal of the CBMs from Xyn10A strongly reduced the ability of plant cell wall hydrolysis. These results suggested that the CBMs of Xyn10A play an important role in the hydrolysis of plant cell walls.

Study on water pollution measurement and integration monitoring system with B-TOX (B-TOX를 이용한 오염수 측정 및 통합관리시스템 구현에 관한 연구)

  • Yang, Jae-Soo;Han, Yong-Hwan;Lee, Young-Ha;Kim, Yoon-Hyun;Kim, Jing-Young
    • The Journal of the Institute of Internet, Broadcasting and Communication
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    • v.10 no.2
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    • pp.61-67
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    • 2010
  • In operating estimation system, the river and lake is almost simple monitoring system or independent monitoring system. But the present system has many weak points about the concurrent analysis ability of various information collection. Sensors generating the information is not accurate. Therefore, we have something that can be improved. Although we have the related facilities of the purification plant, forebays and sewage disposal plant, and suffer from the difficulty about lack of system of estimation, management and control. As a result of this paper, monitoring system can observe the water pollution and various water environment of whole country. Also, through the sensors of USN, Zigbee, RFID and middle ware, which can service and construct service platform, we present standard plan of remarkable scalability service platform.

A Study on Application and Verification of Heavy Water Treatment Effects Using Plant Cultivation (Vegetation) on Floating Island (식생섬에 의한 중수처리 효과 검증과 적용에 관한 연구)

  • Kwon, Dong Min;Kwon, Soon Hyo;Koo, Bon Hak
    • Journal of the Korean Society of Environmental Restoration Technology
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    • v.15 no.6
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    • pp.53-59
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    • 2012
  • This this study was conducted with the aim of doing experiment on the effect of water purification by using an artificially built plant island, which is one of the eco-techniques, and aquatic plants as a plan for the reuse of water for obtaining water resources, thereby analyzing the removed quantity, and applying the experimental results to the reuse of water. As a result of doing experiments, this study obtained a good measured value of BOD (biochemical oxygen demand) 4.7mg/L, and COD (chemical oxygen demand) 7.2mg/L below the heavy water standard of BOD 10mg/L and COD 20mg/L, respectively. The chromaticity showed 89.2% removal efficiency, but final treated wastewater was found to show chromaticity 58 degrees exceeding chromaticity 20 degrees which are the water quality standard of the reuse of water. The results revealed that T-N produced 27% removal efficiency on an average while T-P produced 38% removal efficiency on an average, showing that the removal effect of N & P wasn't big. According to the currently enforced "Water Quality Standard of Heavy Water by Use", the use of water for sprinkling and landscaping was found to be available. Accordingly, this study suggested a nature-friendly, economically-efficient, and eco-technological water treatment technique which will make it possible to overcome the limit of the existing physio-chemical water treatment technology, reduce the costs for maintenance and facilities, and also reduce the limit of space restraint for installation of facilities.

Selective Interaction Between Chloroplast β-ATPase and TGB1L88 Retards Severe Symptoms Caused by Alternanthera mosaic virus Infection

  • Seo, Eun-Young;Nam, Jiryun;Kim, Hyun-Seung;Park, Young-Hwan;Hong, Seok Myeong;Lakshman, Dilip;Bae, Hanhong;Hammond, John;Lim, Hyoun-Sub
    • The Plant Pathology Journal
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    • v.30 no.1
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    • pp.58-67
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    • 2014
  • The multifunctional triple gene block protein 1 (TGB1) of the Potexvirus Alternanthera mosaic virus (AltMV) has been reported to have silencing suppressor, cell-to-cell movement, and helicase functions. Yeast two hybrid screening using an Arabidopsis thaliana cDNA library with TGB1 as bait, and co-purification with TGB1 inclusion bodies identified several host proteins which interact with AltMV TGB1. Host protein interactions with TGB1 were confirmed by biomolecular fluorescence complementation, which showed positive TGB1 interaction with mitochondrial ATP synthase delta' chain subunit (ATP synthase delta'), light harvesting chlorophyll-protein complex I subunit A4 (LHCA4), chlorophyll a/b binding protein 1 (LHB1B2), chloroplast-localized IscA-like protein (ATCPISCA), and chloroplast ${\beta}$-ATPase. However, chloroplast ${\beta}$-ATPase interacts only with $TGB1_{L88}$, and not with weak silencing suppressor $TGB1_{L88}$. This selective interaction indicates that chloroplast ${\beta}$-ATPase is not required for AltMV movement and replication; however, TRV silencing of chloroplast ${\beta}$-ATPase in Nicotiana benthamiana induced severe tissue necrosis when plants were infected by AltMV $TGB1_{L88}$ but not AltMV $TGB1_{L88}$, suggesting that ${\beta}$-ATPase selectively responded to $TGB1_{L88}$ to induce defense responses.

Impact of Pig Slurry from WangGoong Farm Cluster on Water Quality of the Iksan Stream

  • Choi, Hong-Lim;Suresh, A.;Babu, T.;Mansyur, Mansyur
    • Journal of Animal Science and Technology
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    • v.53 no.6
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    • pp.549-561
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    • 2011
  • Livestock wastewater is being discharged without treatment from Hasen's pig farm cluster in WangGoong (WG) area into the Iksan Stream, eventually flowing into the ManGyung (MG) at the upstream junction. Although it is well known that before discharge, wastewater must satisfy the pig slurry discharge standards; because of ongoing remodeling, proper treatment is not being performed. According to public records, wastewater from the WG pig farm cluster is responsible for 3.6% of MG River pollution and 2.0% of the SaeManGuem (SMG) Reservoir pollution. As a result, upstream water treatment quality has become primary concern for development of the SMG project. All physicochemical constituents and pathogenic microbes, such as chemical oxygen demand ($COD_{Cr}$), biochemical oxygen demand ($BOD_5$), total suspended solids (TSS), total nitrogen (TN), total phosphorous (TP), fecal coliforms, Escherichia coli and Salmonella at the effluent of WG Plant (S-1) exceed the effluent standards. This is mainly due to insufficient wastewater treatment: the WG Plant is under renovation to increase water purification efficiency. By comparing the water quality at the S-7 junction, where the the Iksan Stream (pig farms) and the Wanggoong Stream (no pig farms) merge, it is clear that farming facilities and improper treatment can critically affect surrounding water quality. While it is clear throughout this study that the level of all physicochemical parameters and pathogenic microbes along the Stream decreased due to sedimentation, biodegradation and/or dilution. An alarming problem was discovered: the existence of pathogenic microbe count(E coli, Salmonella) in the lagoon wastewater and the stream water. Not only were high concentrations of these pathogens themselves found, but the potential existence of more serious pathogens could rise to more dangerous conditions.

Cellulolytic Enzymes from Acrophialophora nainiana

  • Punnapayak, Hunsa
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2005.06a
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    • pp.245-247
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    • 2005
  • A cellulolytic fungus isolated from Agave plantation in northeastern Thailand was identified as Acrophialophora nainiana. The fungus was capable of growing at pH between 3 - 7 and 25 - 45 $^{\circ}C$, with the optimum conditions at pH 5.0 and 40 $^{\circ}C$. The wild isolate produced cellulases, comprising of exoglucanase (0.019 U/mg protein), endoglucanase (0.366 U/mg protein), and ${\beta}$-glucosidase (0.001 U/mg protein). Mutations with UV and NTG produced the UV 10-2 mutant with cellulases activities including exoglucanase (0.093 U/mg protein), endoglucanase (0.585 U/mg protein), and ${\beta}$-glucosidase (0.013 U/mg protein). Purification of the enzymes with ultrafiltration, ammonium sulfate precipitation, and ion-exchange chromatography yielded the maximal cellulase specific activities of 2.736 U/mg protein (exoglucanase), 0.235 U/mg protein (endoglucanase), and 0.008 U/mg protein (${\beta}$-glucosidase). The mutant's cellulases were the most active at pH 5.0 and 60 $^{\circ}C$. Ion-exchange chromatography revealed that A. nainiana UV 10-2 cellulases were comprised of two peaks with one peak showing the single endoglucanase activity while the other peak showed a mixture of the three enzyme activities. Production of A. nainiana UV 10-2 cellulases using banana leaf stalk as the sole carbon source gave comparable yields to that of the pure ${\alpha}$-cellulose. The enzymes were used in the simultaneous saccharification and fermentation (SSF) of plant residue (Coix aquatica) along with Kluveromyces marxianus to produce ethanol. Moreover, when the enzymes were used in the bioscouring process of fabric, the desiravle traits of textile processing including immediate water absorbency, increased in whiteness and reduction of yellowness of the treated fabric were observed.

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Purification and Partial Characterization of a Peroxidase from Perilla Callus

  • Hur, Yeon-Jae;Lee, Han-Gil;Hu, Gaosheng;Chung, Won-Bok;Jeong, Soon-Jae;Yi, Young-Byong;Nam, Jae-Sung;Chung, Young-Soo;Lee, Jai-Heon;Kim, Doh-Hoon
    • Journal of Plant Biotechnology
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    • v.34 no.4
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    • pp.355-361
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    • 2007
  • Cotyledons of perilla6 were cultured on MS medium containing 0.5 mg/l NAA and 0.5 mg/l BA for 7 weeks. The activity of perilla peroxidase was observed to increase following culture stages as assessed by peroxidase assay. A peroxidase (POD) was purified from perilla tissue cultured on MS medium for 7 weeks. The peroxidase was purified using ion exchange and gel nitration chromatography. The perilla peroxidase had a molecular mass of 30 kDa by SDS-PAGE. We showed that the N-terminal amino acid sequence of this protein shared 67% identity with the tea peroxidase. As indicated by SDS-PAGE, the banding pattern of the 30 kDa polypeptide present in total soluble protein from perilla tissue was increased following culture stages. Immunoblot analysis indicated that perilla peroxidase protein appeared after 3 weeks of perilla tissue culture, and continued to increase with extended duration of tissue culture for at least 7 weeks.

Purification and Characterization of Ice Nucleating Proteins from Ice Nucleation-Active Bacteria (빙핵활성 세균으로부터 빙핵활성 단백질의 정제 및 특성)

  • Kim, Ki-Chung;Lee, Ung;Song, Dong-Up;Cho, Baik-Ho
    • Korean Journal Plant Pathology
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    • v.12 no.1
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    • pp.99-108
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    • 1996
  • 3종의 빙핵세균 Peudomonas syringae 8401, Pseudomonas fuorescens 8701, Erwinia herbicola 8701의 세포 외막으로부터 아무런 변성제도 사용치 않고 sucrose density gradient centrifugation, Sephacryl gel filtration chromatography, DEAE-cellulose ion exchange chromatography, non-denaturing buffer를 이용한 PAGE, electroelution, SDS-PAGE를 통해 빙핵활성 단백질을 고도로 정제할 수 있었다. P. suringae와 P. fluorescens에서는 각각 3종류(155 kD, 75 kD, 50 kD)의 빙핵활성 단백질이, E. herbicola에서는 155 kD를 제외한 2종류(75 kD, 50 kD)의 빙핵활성 단백질은 이 연구를 통해 처음 밝혀진 것으로 , 지금까지 보고된 빙핵활성 단백질(150 KD 이상)보다는 훨씬 작은 것이다. 이는 빙핵활성을 나타내는 단백질의 기본단위는 이 실험의 결과만에 의하면 최대 50 kD임을 시사한다. 이들 단백질은 그 유래된 세균의 종류나 또는 단백질 분자량의 크기에 관계없이 모두 -5.5~7.5$^{\circ}C$에서 물을 동결시키는 높은 빙핵활성을 갖고 있었다. 이는 지금까지 보고된 어느 정제단백질의 빙핵활성보다 높은 것이다. 정제된 단백질의 빙핵활성은 trypsin 처리에 의해 상실되었고, pH6~8범위에서는 안정하였으며, pH5이하, pH9이상에서는 활성을 상실하였다. 보존온도에 대한 영향은 3$0^{\circ}C$이상이 되면 점차 활성이 감소하는 경향을 보이다 37$^{\circ}C$이상에서는 활성이 완전히 상실되었다. 금속이온으로서 Hg\ulcorner이온과 SDS에 의해 활성이 상실되었으나 phosphatidylinositol의 첨가에 의해서는 활성이 약간 증가(-1$^{\circ}C$)하였다.

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Purification and Properties of Homoseine Dehydrogenases in Canavalia Iineata Cotyledons (해녀콩(Canavalia lineata) 자엽에서 Homoserine Dehydrogenase의 정제 및 특성)

  • 최영명
    • Journal of Plant Biology
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    • v.39 no.1
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    • pp.41-48
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    • 1996
  • Two forms of homo serine dehydrogenase have been isolated from 8-day-old cotyledons of Canavalin lineata by a heat denaturation, ammonium sulfate fractionation, DEAE-8ephacel ion exchange and Sephacryl 8-300 gel filtration chromatographies, and Pro cion red dye, Cibacron blue dye and Resource Q column chromatographies. The molecular weights of T -form (threonine-sensitive) and K-form(threonine- insensitive) were estimated to 230 kD and 135 kD, respectively. In the presence of 10 mM threonine, the activity of T-form was inhibited with almost 70%, but that of K-form was not at all. The Km values tor homo serine of T- and Kform were 1.6 mM and 0.3 mM, respectively. The Km values for NAD of T- and K-form were 2.34 mM and 0.03 mM, respectively. And Km values for NADP of two isozymes were the same as 0.01 mM. The activities of T- and K-form were markedly stimulated up to 4.9and 2.8-fold, respectively, by 400 mM KCI. The partial purified(gel filtration) enzymes(Tform and K-form) can be reversibly converted.verted.

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Isolation and Characterization of Endocellulase-Free Multienzyme Complex from Newly Isolated Thermoanaerobacterium thermosaccharolyticum Strain NOI-1

  • Chimtong, Suphavadee;Tachaapaikoon, Chakrit;Pason, Patthra;Kyu, Khin Lay;Kosugi, Akihiko;Mori, Yutaka;Ratanakhanokchai, Khanok
    • Journal of Microbiology and Biotechnology
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    • v.21 no.3
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    • pp.284-292
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    • 2011
  • An endocellulase-free multienzyme complex was produced by a thermophilic anaerobic bacterium, Thermoanaerobacterium thermosaccharolyticum strain NOI-1, when grown on xylan. The temperature and pH optima for growth were $60^{\circ}C$ and 6.0, respectively. The bacterial cells were found to adhere to insoluble xylan and Avicel. A scanning electron microscopy analysis showed the adhesion of xylan to the cells. An endocellulase-free multienzyme complex was isolated from the crude enzyme of strain NOI-1 by affinity purification on cellulose and Sephacryl S-300 gel filtration. The molecular mass of the multienzyme complex was estimated to be about 1,200 kDa. The multienzyme complex showed one protein on native PAGE, one xylanase on a native zymogram, 21 proteins on SDS-PAGE, and 5 xylanases on a SDS zymogram. The multienzyme complex consisted of xylanase, ${\beta}$-xylosidase, ${\alpha}$-L-arabinofuranosidase, ${\beta}$-glucosidase, and cellobiohydrolase. The multienzyme complex was effective in hydrolyzing xylan and corn hulls. This is the first report of an endocellulase-free multienzyme complex produced by a thermophilic anaerobic bacterium, T. thermosaccharolyticum strain NOI-1.