• Title/Summary/Keyword: Activity Sequencing

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Successful start-up of pilot-scale single-stage ANAMMOX reactor through cultivation of ammonia oxidizing and ANAMMOX bacteria (암모니아 산화균 및 아나목스균의 배양을 통한 파일롯 규모 단일 아나목스 반응기의 성공적인 시운전)

  • Choi, Daehee;Jin, Yangoh;Lee, Chulwoo;Jung, Jinyoung
    • Journal of Korean Society of Water and Wastewater
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    • v.32 no.5
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    • pp.371-379
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    • 2018
  • The lack of seed sludges for Ammonium Oxidizing Bacteria (AOB) and slow-growing ANaerobic AMMonium OXidation (ANAMMOX) bacteria is one of the major problem for large-scale application. In this study, $24m^3$ of single-stage SBR (Sequencing Batch Reactor) was operated to remove nitrogen from reject water using AOB and ANAMMOX bacteria cultivated from activated sludge in the field. The ANAMMOX activity was found after 44 days of cultivation in the ANAMMOX cultivation reactor, and then $0.66kg\;N/m^3/d$ of the nitrogen removal rate was achieved at $0.78kg\;N/m^3/d$ of the nitrogen loading rate at 153 days of cultivation. The AOB cultivation reactor showed $0.2kg\;N/m^3/d$ of nitrite production rate at $0.4kg\;N/m^3/d$ of nitrogen loading rate after 36 days of operation. The cultivated ANAMMOX bacteria and AOB was mixed into the single-stage SBR. The feed distribution was applied to remove total nitrogen stably in the single-stage SBR. The nitrogen removal rate in the single-stage SBR was gradually enhanced with an increase of specific activities of both AOB and ANAMMOX bacteria by showing $0.49kg\;N/m^3/d$ of the nitrogen removal rate at $0.56kg\;N/m^3/d$ of the nitrogen loading rate at 54 days of operation.

Candidate of Probiotic Bacteria Isolated from Several Jeotgals: Korean Traditional Fermented Seafoods

  • Cho, Gyu-Sung;Do, Hyung-Ki;Bae, Chae-Yoon;Cho, Gyu-Sup;Whang, Cher-Won;Shin, Heuyn-Kil
    • Preventive Nutrition and Food Science
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    • v.11 no.2
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    • pp.140-145
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    • 2006
  • Seventy eight bacterial strains were isolated from several jeotgals using MRS and M 17 agar media. The probiotic properties such as tolerance of extreme growth condition, production of antimicrobial compound, production of hydrogen peroxide, and enzymatic activity of bile salt hydrolase were investigated. DHK 4, 10, 21 and 74 strains showed_a strong tolerance property against extreme conditions such as low pH and 0.5% oxgall-supplemented medium. DHK 10 and 47 strains produced hydrogen peroxide on TMB agar plate. DHK 8 and 10 strains produced antimicrobial compounds onto MRS agar against E. facalis. DHK 4, 6, 21, 29, 33, 63 and 87 strains had high activities of bile salt hydrolase. Especially, DHK 10 displayed a strong probiotic candidate; the abilities to produce the antimicrobial compound, hydrogen peroxide, and bile salt hydrolase. All these strains are assumed to be useful probiotic candidates. Among 78, twenty seven strains which have probiotic properties were tentatively identified by 16S rRNA sequencing. Among them, 7 Lactobacillus spp., 6 Leuconosotoc spp., 2 Weisella spp., 1 Pediococcus sp., 1 Staphylococcus sp., 1 Enterococcus sp. and 2 Streptococcus spp. were tentatively identified.

Characteristics of Aerobic Granular Activated Sludge According to Electron Acceptors in Sequencing Batch Reactor Process (SBR공정에서 전자수용체에 따른 호기성 입상활성슬러지의 공정별 특성)

  • Kim, I-Tae;Lee, Hee-Ja;Bae, Woo-Keun
    • Journal of Korean Society on Water Environment
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    • v.20 no.5
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    • pp.480-487
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    • 2004
  • This study was conducted to find the effect of electron acceptors on the formation of granular sludge by using four different types of electron acceptors. The phosphorous uptake, denitrification, and sulfate reduction in anoxic modes were simultaneously occured because of the presence of the polyphosphate accumultating organism(PAO) that utilize nitrate and sulfate as an electron acceptor in the anoxic zone. Denitrirying phosphorous removal bacteria(DPB) was enriched under anaerobic/anoxic/aerobic condition with a nitrate as an electron acceptor, and desulfating phosphorous removal bacteria(DSPB) was enriched under anaerobic/anoxic/aerobic condition with a sulfate as an electron acceptor. Polyphosphate accumulating organism(PAO) were enriched in the anaerobic/aerobic SBR. PAO took up acetate faster than DPB and DSPB during the aerobic phase. The sludge with nitrate and sulfate as an electron acceptors grew as a granules which possessed high activity and good settleability. In the anaerobic/aerobic modes, typical floccular growth was observed. In the result of bench-scale experiment, simultaneous reactions of phosphorus uptake, denitrification and sulfate reduction were observed under anoxic condition with nitrate and sulfate as an electron acceptors. These results demonstrated that the anaerobic/anoxic modes with nitrate and sulfate as an electron acceptors played an important role in the formation of the sludge granulation.

Identification by 16S rDNA Partial Sequencing of Thermophilic Bacteria with Fermentation of Pig Manure (16S rDNA 증폭에 의한 부분염기서열을 이용한 분뇨 발효 관련 고온 호기성 박테리아의 동정)

  • Kim, Myung Kil;Choi, Don Ha;Choi, In Gyu;Kim, Byung Gyu;Song, Jae Gyung
    • Journal of the Korean Wood Science and Technology
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    • v.34 no.1
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    • pp.68-78
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    • 2006
  • The purpose of this study was to estimate the identification of thermophilic hacterial with fermentation of pig manure. To identify the characters of thermophilic bacteria related to fermentation at a high temperature condition, we selected 28 different kinds of original settling thermophilic bacteria that were sampled at different 23 areas. They were distributed 1$1{\times}10^5{\sim}10^8CFU$ at medium and the enzyme activity at $55^{\circ}C$ incubation condition, especially cellulase and a-amylase, were higher than those of $30^{\circ}C$. Partial sequencing data for 165 rDNA region were obtained from 28 samples representing 15 different genera. Bacillus subcilis, one of those bacteria, has endodermic spores at high fermented condition.

Effects of Fermented Leachate of Food Waste (FLFW) and Temperature on Nutrient Removal in Sequencing Batch Reactor

  • Roh, Sung-Hee;Chun, Young-Nam;Lee, Sook-Young;Cheong, Hyeon-Sook;Lee, Jae-Wook;Kim, Sun-Il
    • Environmental Engineering Research
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    • v.13 no.3
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    • pp.155-161
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    • 2008
  • This study examined effects of the fermented leachate of food waste (FLFW) on nitrogen and phosphorous removal for domestic wastewater containing a low carbon-to-nitrogen (C/N) ratio in sequencing batch reactor (SBR). When the FLFW was not supplied in the process, release of phosphorus and excessive intake was not observed at both anaerobic and aerobic stages. On the other hand, when the FLFW was gradually added, active release of phosphorus and intake of phosphorus was noticed at an anaerobic stage and aerobic stage, respectively, resulting in improved phosphorus removal efficiency. The removal efficiency of nitrogen and phosphorus was increased from 75% and 37% (R-1, control test) to 97% and 80% (R-4, the highest substrate ratio test), respectively. In addition, although activity of the nitrogen oxidizing microorganisms was reduced when the reaction temperature was decreased to $10^{\circ}C$, the phosphorus removal efficiency was shown to increase with the addition of FLFW, indicating an independence from temperature. Overall, this study suggests that an efficient nutrients removal process can be successfully employed into a SBR when the FLFW is added to a wastewater which has a low C/N ratio.

High-rate Denitrifying Process Based on Methanol and Characteristics of Organic Carbon Uptake (메탄올 기반 탈질 공정의 고속화 및 탄소 섭취 특성)

  • Park, Suin;Jeon, Junbeom;Bae, Hyokwan
    • Journal of Korean Society on Water Environment
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    • v.36 no.6
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    • pp.581-591
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    • 2020
  • In this study, two types of reactors were operated to examine the properties of methanol uptake under the high-rate denitrification process. In a sequencing batch reactor, the denitrifying activity was enriched up to 0.80 g-N/g-VSS-day for 72 days. Then, the enriched denitrifying sludge was transferred to a completely stirred tank reactor (CSTR). At the final phase on Day 46-50, the nitrogen removal efficiency was around 100% and the total nitrogen removal rate reached 0.097±0.003 kg-N/㎥-day. During the continuous process, the sludge settling index (SVI30) was stabilized as 118.3 mL/g with the biomass concentration of 1,607 mg/L. The continuous denitrifying process was accelerated by using a sequencing batch reactor (SBR) with a total nitrogen removal rate of 0.403±0.029 kg-N/㎥-day with a high biomass concentration of 8,433 mg-VSS/L. Because the reactor was open to ambient air with the dissolved oxygen range of 0.2-0.5 mg-O2/L, an increased organic carbon requirement of 5.58±0.70 COD/NO3--N was shown for the SBR in comparison to the value of 4.13±0.94 for the test of the same biomass in a completely anaerobic batch reactor. The molecular analysis based on the 16S rRNA gene showed that Methyloversatilis discipulorum and Hyphomicrobium zavarzinii were the responsible denitrifiers with the sole organic carbon source of methanol.

Modification of ginsenoside saponin composition via the CRISPR/Cas9-mediated knockout of protopanaxadiol 6-hydroxylase gene in Panax ginseng

  • Choi, Han Suk;Koo, Hyo Bin;Jeon, Sung Won;Han, Jung Yeon;Kim, Joung Sug;Jun, Kyong Mi;Choi, Yong Eui
    • Journal of Ginseng Research
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    • v.46 no.4
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    • pp.505-514
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    • 2022
  • Background: The roots of Panax ginseng contain two types of tetracyclic triterpenoid saponins, namely, protopanaxadiol (PPD)-type saponins and protopanaxatiol (PPT)-type saponins. In P. ginseng, the protopanaxadiol 6-hydroxylase (PPT synthase) enzyme catalyses protopanaxatriol (PPT) production from protopanaxadiol (PPD). In this study, we constructed homozygous mutant lines of ginseng by CRISPR/Cas9-mediated mutagenesis of the PPT synthase gene and obtained the mutant ginseng root lines having complete depletion of the PPT-type ginsenosides. Methods: Two sgRNAs (single guide RNAs) were designed for target mutations in the exon sequences of the two PPT synthase genes (both PPTa and PPTg sequences) with the CRISPR/Cas9 system. Transgenic ginseng roots were generated through Agrobacterium-mediated transformation. The mutant lines were screened by ginsenoside analysis and DNA sequencing. Result: Ginsenoside analysis revealed the complete depletion of PPT-type ginsenosides in three putative mutant lines (Cr4, Cr7, and Cr14). The reduction of PPT-type ginsenosides in mutant lines led to increased accumulation of PPD-type ginsenosides. The gene editing in the selected mutant lines was confirmed by targeted deep sequencing. Conclusion: We have established the genome editing protocol by CRISPR/Cas9 system in P. ginseng and demonstrated the mutated roots producing only PPD-type ginsenosides by depleting PPT-type ginsenosides. Because the pharmacological activity of PPD-group ginsenosides is significantly different from that of PPT-group ginsenosides, the new type of ginseng mutant producing only PPD-group ginsenosides may have new pharmacological characteristics compared to wild-type ginseng. This is the first report to generate target-induced mutations for the modification of saponin biosynthesis in Panax species using CRISPR-Cas9 system.

Minoxidil Regulates Aging-Like Phenotypes in Rat Cortical Astrocytes In Vitro

  • Minji Bang;Seung Jin Yang;TaeJin Ahn;Seol-Heui Han;Chan Young Shin;Kyoung Ja Kwon
    • Biomolecules & Therapeutics
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    • v.31 no.1
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    • pp.116-126
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    • 2023
  • Mainly due to the slanted focus on the mechanism and regulation of neuronal aging, research on astrocyte aging and its modulation during brain aging is scarce. In this study, we established aged astrocyte culture model by long-term culturing. Cellular senescence was confirmed through SA-β-gal staining as well as through the examination of morphological, molecular, and functional markers. RNA sequencing and functional analysis of astrocytes were performed to further investigate the detailed characteristics of the aged astrocyte model. Along with aged phenotypes, decreased astrocytic proliferation, migration, mitochondrial energetic function and support for neuronal survival and differentiation has been observed in aged astrocytes. In addition, increased expression of cytokines and chemokine-related factors including plasminogen activator inhibitor -1 (PAI-1) was observed in aged astrocytes. Using the RNA sequencing results, we searched potential drugs that can normalize the dysregulated gene expression pattern observed in long-term cultured aged astrocytes. Among several candidates, minoxidil, a pyrimidine-derived anti-hypertensive and anti-pattern hair loss drug, normalized the increased number of SA-β-gal positive cells and nuclear size in aged astrocytes. In addition, minoxidil restored up-regulated activity of PAI-1 and increased mitochondrial superoxide production in aged astrocytes. We concluded that long term culture of astrocytes can be used as a reliable model for the study of astrocyte senescence and minoxidil can be a plausible candidate for the regulation of brain aging.

Evaluation and Genome Mining of Bacillus stercoris Isolate B.PNR1 as Potential Agent for Fusarium Wilt Control and Growth Promotion of Tomato

  • Rattana Pengproh;Thanwanit Thanyasiriwat;Kusavadee Sangdee;Juthaporn Saengprajak;Praphat Kawicha;Aphidech Sangdee
    • The Plant Pathology Journal
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    • v.39 no.5
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    • pp.430-448
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    • 2023
  • Recently, strategies for controlling Fusarium oxysporum f. sp. lycopersici (Fol), the causal agent of Fusarium wilt of tomato, focus on using effective biocontrol agents. In this study, an analysis of the biocontrol and plant growth promoting (PGP) attributes of 11 isolates of loamy soil Bacillus spp. has been conducted. Among them, the isolates B.PNR1 and B.PNR2 inhibited the mycelial growth of Fol by inducing abnormal fungal cell wall structures and cell wall collapse. Moreover, broad-spectrum activity against four other plant pathogenic fungi, F. oxysporum f. sp. cubense race 1 (Foc), Sclerotium rolfsii, Colletotrichum musae, and C. gloeosporioides were noted for these isolates. These two Bacillus isolates produced indole acetic acid, phosphate solubilization enzymes, and amylolytic and cellulolytic enzymes. In the pot experiment, the culture filtrate from B.PNR1 showed greater inhibition of the fungal pathogens and significantly promoted the growth of tomato plants more than those of the other treatments. Isolate B.PNR1, the best biocontrol and PGP, was identified as Bacillus stercoris by its 16S rRNA gene sequence and whole genome sequencing analysis (WGS). The WGS, through genome mining, confirmed that the B.PNR1 genome contained genes/gene cluster of a nonribosomal peptide synthetase/polyketide synthase, such as fengycin, surfactin, bacillaene, subtilosin A, bacilysin, and bacillibactin, which are involved in antagonistic and PGP activities. Therefore, our finding demonstrates the effectiveness of B. stercoris strain B.PNR1 as an antagonist and for plant growth promotion, highlighting the use of this microorganism as a biocontrol agent against the Fusarium wilt pathogen and PGP abilities in tomatoes.

Transcriptome and proteome analysis of pregnancy and postpartum anoestrus ovaries in yak

  • Chen, Zhou;Wang, Jine;Ma, Junyuan;Li, Shuyuan;Huo, Shengdong;Yang, Yanmei;Zhaxi, Yingpai;Zhao, Yongqing;Zhang, Derong
    • Journal of Veterinary Science
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    • v.23 no.1
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    • pp.3.1-3.12
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    • 2022
  • Background: Domestic yaks are the most important livestock species on the Qinghai-Tibetan Plateau. Adult female yaks normally breed in the warm season (July to September) and enter anestrous in the cold season (November to April). Nevertheless, it is unclear how ovarian activity is regulated at the molecular level. Objectives: The peculiarities of yak reproduction were assessed to explore the molecular mechanism of postpartum anestrus ovaries in yaks after pregnancy and parturition. Methods: Sixty female yaks with calves were observed under natural grazing in Haiyan County, Qinghai Province. Three yak ovaries in pregnancy and postpartum anestrus were collected. RNA sequencing and quantitative proteomics were employed to analyze the pregnancy and postpartum ovaries after hypothermia to identify the genes and proteins related to the postpartum ovarian cycle. Results: The results revealed 841 differentially expressed genes during the postpartum hypoestrus cycle; 347 were up-regulated and 494 genes were down-regulated. Fifty-seven differential proteins were screened: 38 were up-regulated and 19 were down-regulated. The differential genes and proteins were related to the yak reproduction process, rhythm process, progesterone-mediated oocyte maturation, PI3K/AKT signaling pathway, and MAPK signaling pathway categories. Conclusions: Transcriptome and proteomic sequencing approaches were used to investigate postpartum anestrus and pregnancy ovaries in yaks. The results confirmed that BHLHE40, SF1IX1, FBPX1, HSPCA, LHCGR, BMP15, and ET-1R could affect postpartum hypoestrus and control the state of estrus.