• Title/Summary/Keyword: $NH_3-NH_4^+$

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Biochemical Characterization of Recombinant L-Asparaginase (AnsA) from Rhizobium etli, a Member of an Increasing Rhizobial-Type Family of L-Asparaginases

  • Moreno-Enriquez, Angelica;Evangelista-Martinez, Zahaed;Gonzalez-Mondragon, Edith G.;Calderon-Flores, Arturo;Arreguin, Roberto;Perez-Rueda, Ernesto;Huerta-Saquero, Alejandro
    • Journal of Microbiology and Biotechnology
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    • v.22 no.3
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    • pp.292-300
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    • 2012
  • We report the expression, purification, and characterization of L-asparaginase (AnsA) from Rhizobium etli. The enzyme was purified to homogeneity in a single-step procedure involving affinity chromatography, and the kinetic parameters $K_m$, $V_{max}$, and $k_{cat}$ for L-asparagine were determined. The enzymatic activity in the presence of a number of substrates and metal ions was investigated. The molecular mass of the enzyme was 47 kDa by SDS-PAGE. The enzyme showed a maximal activity at $50^{\circ}C$, but the optimal temperature of activity was $37^{\circ}C$. It also showed maximal and optimal activities at pH 9.0. The values of $K_m$, $V_{max}$, $k_{cat}$, and $k_{cat}/K_m$ were $8.9{\pm}0.967{\times}10^{-3}$ M, $128{\pm}2.8$ U/mg protein, $106{\pm}2s^{-1}$, and $1.2{\pm}0.105{\times}10^4M^{-1}s^{-1}$, respectively. The L-asparaginase activity was reduced in the presence of $Mn^{2+}$, $Zn^{2+}$, $Ca^{2+}$, and $Mg^{2+}$ metal ions for about 52% to 31%. In addition, we found that $NH_4{^+}$, L-Asp, D-Asn, and ${\beta}$-aspartyl-hydroxamate in the reaction buffer reduced the activity of the enzyme, whereas L-Gln did not modify its enzymatic activity. This is the first report on the expression and characterization of the L-asparaginase (AnsA) from R. etli. Phylogenetic analysis of asparaginases reveals an increasing group of known sequences of the Rhizobial-type asparaginase II family.

Comparison of Biochemical Characterization of Korean and Chinese Mung Bean Lectin (한국산 녹두와 중국산 녹두에 있어서 Lectin의 생화학적 특성 비교)

  • Roh, Kwang Soo
    • Journal of Life Science
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    • v.24 no.6
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    • pp.603-611
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    • 2014
  • The lectins were separated from Korean and Chinese mung bean seeds finally via chromatography using Sephadex G-100 and their biochemical features were studied and compared. They showed no hemagglutination with human red blood cells regardless of trypsin treatment and showed hemagglutination with only trypsin treated rabbit red blood cells. The molecular weights of two lectins were identified as 54 kDa and 28 kDa by SDS-PAGE. It was found that while the optimal reaction temperature of the lectin from Korean mung bean was $60^{\circ}C$, that of the lectin from Chinese mung bean seeds was $50^{\circ}C$. It was found also that the most thermal stable temperature of the seed lectin from Korean mung bean seeds was $50^{\circ}C$ and the lectin from Chinese mung bean was $40-50^{\circ}C$. The lectin from Korean mung bean seeds showed the highest activity at pH 3.2 and the lectin from Chinese mung bean showed the highest activity at pH 6.2. It was identified that when treating a denaturant, thiourea and guanidine-HCl resulted in no hemagglutination, so they induced denaturalization. It was identified also that there was no hemagglutination with urea, so it did not induced denaturalization. They showed no septicity to 6 types of carbohydrates including D-glucose. In addition, the lectins from the two mung bean seed had specificity to metal ions.

The improvement of ginsenoside accumulation in Panax ginseng as a result of γ-irradiation

  • Kim, Dong Sub;Song, Mira;Kim, Sun-Hee;Jang, Duk-Soo;Kim, Jin-Baek;Ha, Bo-Keun;Kim, Sang Hoon;Lee, Kyung Jun;Kang, Si-Yong;Jeong, Il Yun
    • Journal of Ginseng Research
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    • v.37 no.3
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    • pp.332-340
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    • 2013
  • In this study, gamma rays were used to irradiate embryogenic calli induced from cotyledon explants of Panax ginseng Meyer. After the embryogenic calli were irradiated, they were transferred to adventitious roots using an induction medium; next, mutated adventitious root (MAR) lines with a high frequency of adventitious root formations were selected. Two MAR lines (MAR 5-2 and MAR 5-9) from the calli treated with 50 Gy of gamma rays were cultured on an $NH_4NO_3$-free Murashige and Skoog medium with indole-3-butyric acid 3 mg/L. The expression of genes related to ginsenoside biosynthesis was analyzed using reverse transcription polymerase chain reaction with RNA prepared from native ginseng (NG), non-irradiated adventitious root (NAR) and 2 MAR lines. The expression of the squalene epoxidase and dammarenediol synthase genes was increased in the MAR 5-2 line, whereas the phytosterol synthase was increased in the MAR 5-9 line. The content and pattern of major ginsenosides (Rb1, Rb2, Rc, Rd, Re, Rf, and Rg1) were analyzed in the NG, NAR, and 2 MAR lines (MAR 5-2 and MAR 5-9) using TLC and HPLC. In the TLC analysis, the ginsenoside patterns in the NG, NAR, and 2 MAR lines were similar; in contrast, the MAR 5-9 line showed strong bands of primary ginsenosides. In the HPLC analysis, compared with the NG, one new type of ginsenoside was observed in the NAR and 2 MAR lines, and another new type of ginsenoside was observed in the 2 MAR lines irradiated with gamma rays. The ginsenoside content of the MAR 5-9 line was significantly greater in comparison to the NG.

Mass Reduction and Physicochemical Properties of the Produced Compost during Composting Domestic Food Wastes in a Small Composter (소형 퇴비화용기에서 가정 음식물쓰레기의 퇴비화 과정 중 감량화 및 생산 퇴비의 물리화학적 특성)

  • Park, Ju-Won;Seo, Jeoung-Yoon
    • Korean Journal of Environmental Agriculture
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    • v.20 no.4
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    • pp.238-243
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    • 2001
  • Mass reduction and physicochemical properties of the produced compost were investigated during composting domestic food wastes without additive. A small composter used in this study had the height of 15 cm from the center of bottom half circle (diameter 24 cm) up to under the lid, the side length of 50 cm and the horizontal lid angle of $50^{\circ}$ and was operated at the heating unit temperature of $85^{\circ}$. It was mixed by the rotating arm for two minutes in every half hour while supplied with air flow at 3 L/min for 10 minutes in every half hour. This condition was found in a preliminary experiment as optimal for keeping the water content of composting material in the optimal range without adding any bulking materials. The domestic food wastes were added into the composter at the rate of 1 kg/day without additives during composting. The results were as follows; during the composting process, water content maintained in the range of $51.0{\sim}53.5%$. Hemicellulose and lignin contents did not show any tendency, but cellulose content decreased. During the composting process, $NH_3-N$ and $NO_2-N$ were not detected due to nitrification. The contents of inorganic compounds did not increase during the composting process. They were in the range of $1.32{\sim}1.71%\;P_2O_5$, $1.29{\sim}1.48%\;CaO$, $0.41{\sim}0.49%\;MgO$, and $0.38{\sim}0.74%\;K_2O$. For 20 days, weight reduction rate was 67.5% in wet basis, and decomposition rate was 48% in dry basis. Concentration of heavy metals (Cu, Cr, Cd, Pb, Zn, Hg, As) was less than the limiting value of the compost. Maturity of the produced compost was 3 grade through reaching maximum temperature of $46{\sim}48^{\circ}C$.

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Temporal and Spatial Evaluation of Water Pollution Characteristics in Gohyeon Stream and Its Tributaries (고현천 및 유입지류의 수질오염 특성의 시·공간적 평가)

  • Kim, Sung-Jae
    • Korean Journal of Environmental Agriculture
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    • v.31 no.3
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    • pp.235-247
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    • 2012
  • BACKGROUND: Gohyeon Stream is the municipal eco-stream of 7.1km in total length which flows through the downtown area of Gohyeon in Geoje city, rising from the watershed of Mundong Water Fall. Gohyeon district in Geoje city has been a rapid growing area centering in Geoje city and then experienced an rapid increase in population. Large amounts of sewage pollutants have been spewed into Gohyeon Stream from its tributaries, due to the lack of sewer system. Gohyeon Stream is laced with unhealthy levels of fecal coliform (FC). Restoration of water quality in Gohyeon Stream is no less inevitable in behalf of its ecosystem and the citizen. In this study, the water quality of Gohyeon Stream and its tributaries was examined temporally and spatially, and their relationships were comparatively analyzed to give useful basic data applying to a restoration project of the water quality of Gohyeon Stream. METHODS AND RESULTS: The samples ware taken at 20 points in Gohyeon Stream and 19 points in its tributaries during the rainy and dry seasons, respectively, and examined on the parameters of pH, temperature, salinity, dissolved oxygen (DO), suspended solid (SS), biochemical oxygen demand (BOD), chemical oxygen demand (COD), dissolved inorganic nitrogen (DIN; $NH_3$-N, $NO_3$-N, $NO_2$-N), disolved inorganic phosphorus (DIP; $PO_4$-P) and FC. The data were analyzed using a comparative analysis and Pearson's correlation analysis among the parameters. During the rainy season, the concentration of SS was high in the upper region of Gohyeon Stream, and the concentrations of COD, DIN and DIP were low in the upper region and high in the middle and lower regions. During the dry season, the concentration of SS was low and the concentrations of COD, DIN, DIP and FC were high in all regions. The Pearson's correlation analyses showed that the relationships between DO and FC, COD and DIP, and DIN and FC during the rainy season as well as between DO and DIN, SS and FC, COD and DIP, and DIN and DIP during the dry season were significant. CONCLUSION: During the rainy season, the upper region of Gohyeon Stream flowed the high level of SS, the middle region the high level of nutrients due to an agricultural run-off, and the lower region the high level of nutrients due to a sewage inflow. During the dry season, the water quality of Gohyeon Stream was directly and sensitively influenced on the inflow of sewage from the tributaries.

The effectiveness of step feeding strategies in sequencing batch reactor for a single-stage deammonification of high strength ammonia wastewater

  • Choi, Wonyoung;Yu, Jaecheul;Kim, Jeongmi;Jeong, Soyeon;Direstiyani, Lucky Caesar;Lee, Taeho
    • Membrane and Water Treatment
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    • v.11 no.1
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    • pp.79-85
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    • 2020
  • A single-stage deammonification with a sequencing batch reactor (SBR) that simultaneous nitritation, anaerobic ammonia oxidation (anammox), and denitrification (SNAD) occur in one reactor has been widely applied for sidestream of wastewater treatment plant. For the stable and well-balanced SNAD, a feeding strategy of influent wastewater is one of the most important operating factors in the single-stage deammonification SBR. In this study, single-stage deammonification SBR (working volume 30L) was operated to treat a high-strength ammonium wastewater (1200 mg NH4+-N/L) with different feeding strategies (single feeding and nine-step feeding) under the condition without COD. Each cycle of the step feeding involved 6 sub-cycles consisted of aerobic and anoxic periods for partial nitritation (PN) and anammox, respectively. Contrary to unstable performance in the single feeding, the step feeding showed better deammonification performance (0.565 kg-N/m3/day). Under the condition with COD, however, the nitrogen removal rate (NRR) decreased to 0.403 kg-N/m3/day when the Nine-step feeding strategies had an additional denitrification period before sub-cycles for PN and anammox. The NRR was recovered to 0.518 kg-N/m3/day by introducing an enhanced multiple-step feeding strategy. The strategy had 50 cycles consisted of feed, denitrification, PN, and anammox, instead of repeated sub-cycles for PN and anammox. The multiple-step feeding strategy without sub-cycle showed the most stable and excellent deammonification performance: high nitrogen removal efficiency (98.6%), COD removal rate (0.131 kg-COD/m3/day), and COD removal efficiency (78.8%). This seemed to be caused by that the elimination of the sub-cycles might reduce COD oxidation during aerobic condition but increase the COD utilization for denitrification period. In addition, among various sensor values, the ORP pattern appeared to be applicable to monitor and control each reaction step for deammonification in the multiple-step feeding strategy without sub-cycle. Further study to optimize the number of multiple-step feeding is still needed but these results show that the multiple-step feeding strategy can contribute to a well-balanced SNAD for deammonification when treating high-strength ammonium wastewater with COD in the single-stage deammonification SBR.

Ensiled Green Tea Waste as Partial Replacement for Soybean Meal and Alfalfa Hay in Lactating Cows

  • Kondo, Makoto;Nakano, Masashi;Kaneko, Akemi;Agata, Hirobumi;Kita, Kazumi;Yokota, Hiroomi
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.7
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    • pp.960-966
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    • 2004
  • The purpose of this study was to evaluate the effects of protein supplementation of green tea waste (GTW) on the performance of lactating cows. Another aim was to increase resource utilization and to eliminate any environmental negative impact from the tea waste. GTW from a beverage company was ensiled at a low pH (<4.0) and high acetic acid and lactic acid concentration, and it contained high crude protein (CP, 34.8%), total extractable tannins (TET, 9.2%) and condensed tannin (CT, 1.7%). Two experiments were conducted to investigate the palatability and performance in lactating cows fed GTW. In the palatability trial, three lactating cows were allocated to three dietary treatments in a 3$\times$3 Latin square design. The animals were offered a total mixed ration (TMR) including GTW at rates of 0, 2.5 and 5.0% on a dry matter (DM) basis. Total DM intake was not different among the treatments. In the performance trial, four lactating cows were used in a 2$\times$2 Latin square design with a 3 week sampling period. GTW was incorporated into TMR at a rate of 5.0% on a DM and 10.0% on a CP basis. Thus GTW replaced alfalfa hay and soybean meal at a level of 25.0% on a DM. DM and CP intake were not affected by the inclusion of GTW, whereas TET and CT intake were significantly increased (p<0.001). Milk production, milk composition and the efficiency of milk production were not altered by the GTW inclusion. Although ruminal pH and VFA, and blood urea nitrogen were not changed, ruminal $NH_{3}-N$ and plasma total cholesterol were relatively low in the GTW group, but not significantly different. The excretion of urinary purine derivatives and estimated MN supply were also not significantly affected by GTW treatment. It is therefore concluded that GTW can be used as a protein source without any detrimental effects on the performance of lactating cows.

Nitrogen Removal from Milking Center Wastewater via Simultaneous Nitrification and Denitrification Using a Biofilm Filtration Reactor

  • Won, Seung-Gun;Jeon, Dae-Yong;Kwag, Jung-Hoon;Kim, Jeong-Dae;Ra, Chang-Six
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.6
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    • pp.896-902
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    • 2015
  • Milking center wastewater (MCW) has a relatively low ratio of carbon to nitrogen (C/N ratio), which should be separately managed from livestock manure due to the negative impacts of manure nutrients and harmful effects on down-stream in the livestock manure process with respect to the microbial growth. Simultaneous nitrification and denitrification (SND) is linked to inhibition of the second nitrification and reduces around 40% of the carbonaceous energy available for denitrification. Thus, this study was conducted to find the optimal operational conditions for the treatment of MCW using an attached-growth biofilm reactor; i.e., nitrogen loading rate (NLR) of 0.14, 0.28, 0.43, and $0.58kg\;m^{-3}\;d^{-1}$ and aeration rate of 0.06, 0.12, and $0.24\;m^3\;h^{-1}$ were evaluated and the comparison of air-diffuser position between one-third and bottom of the reactor was conducted. Four sand packed-bed reactors with the effective volume of 2.5 L were prepared and initially an air-diffuser was placed at one third from the bottom of the reactor. After the adaptation period of 2 weeks, SND was observed at all four reactors and the optimal NLR of $0.45kg\;m^{-3}\;d^{-1}$ was found as a threshold value to obtain higher nitrogen removal efficiency. Dissolved oxygen (DO) as one of key operational conditions was measured during the experiment and the reactor with an aeration rate of $0.12\;m^3\;h^{-1}$ showed the best performance of $NH_4-N$ removal and the higher total nitrogen removal efficiency through SND with appropriate DO level of ${\sim}0.5\;mg\;DO\;L^{-1}$. The air-diffuser position at one third from the bottom of the reactor resulted in better nitrogen removal than at the bottom position. Consequently, nitrogen in MCW with a low C/N ratio of 2.15 was successfully removed without the addition of external carbon sources.

Field Application of a Continuously Aerated Bio-Reactor (CABR) for the Treatment of Swine Wastewater (양돈분뇨처리에 있어서 연속폭기배양조(CABR)의 현장적용연구)

  • Lee, Myung-Gyu
    • Journal of Animal Environmental Science
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    • v.1 no.1
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    • pp.55-64
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    • 1995
  • A wastewater purifying system using phototrophic bacterium, Rhodopseudomonas capsulata, is currently in operation in several countries, One of them, is a continuously aerated bioreactor(CABR) system, which treats concentrated swin wasterwater using small amounts of phototrophic bacteria as additive bacterial seeding. Using this plant, total biochemical oxygen demand was decreased to 13%, and most of volatile fatty acids were removed. About 40% of the wastewater(Influx) was evaporated during aerobic digestion for 24h, and 60% of that erupted in a decodorized foam(Efflux). The efflux had enough nutrients, N, P and K kor growing plant, as well as organic matters. When the efflux was applied to Italian ryegrass with high dose, fresh shoot and root weights were significantly greater, and $NO_3-N$ contents of the dried shoot were lower than those of control plant (CDU). These results indicate that the CABR plant is useful for reduction and deodorization of swine wastewater and the efflux from CABR can be used for crop production as an organic fertilizer.

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Study for the Establishment of the Quality Index of Low-salted Myungran-jeot (저염 명란젓갈의 품질지표 설정을 위한 연구)

  • Han Jin-Suk;Cho Hak-Rae;Cho Ho Sung
    • Korean journal of food and cookery science
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    • v.21 no.4 s.88
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    • pp.440-446
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    • 2005
  • To determine the shelf-life of low salted Myungran-jeot (low salted and fermented Alaska pollack roe), various chemical, microbiological and sensory analyses were carried out with Myungran-jeot stored at 10 days. The lactic acid content was slowly increased in the early stage of storage but then was rapidly increased with lengthening storage period, while pH was decreased during storage. The $NH_2-N$ content was steady at 10 days and peaked at 17 days. The production of VBN was increased to over $30mg\%$ at 19 days. In microbiological tests, the viable cell count and halotolerant bacteria were $1.5\times10^3\;and\;5.5\times10^3CFU/g$, respectively, at the initial state, but they slowly increased to $10^7 CFU/g$ at 17 days. Yeast or fungi did not appear on the surface of Myungran-jeot. Sensory evaluation showed that the original torture of Myungran-jeot in the mouth was a very important indicator for acceptability. Sensory characters and acceptability of Myungran-jeot at 17 days were not significantly different from those of the initial state. From these results, the estimated shelf-life of Myungran-jeot whom sealed with aluminum coated paper, was about 18 days in this study.