• Title/Summary/Keyword: $D_2{^+}$ retention

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Tryptophan-Based Hyperproduction of Bioindigo by Combinatorial Overexpression of Two Different Tryptophan Transporters

  • Hyun Jin Kim;Sion Ham;Nara-Shin;Jeong Hyeon Hwang;Suk Jin Oh;Tae-Rim Choi;Jeong Chan Joo;Shashi Kant Bhatia;Yung-Hun Yang
    • Journal of Microbiology and Biotechnology
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    • v.34 no.4
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    • pp.969-977
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    • 2024
  • Indigo is a valuable, natural blue dye that has been used for centuries in the textile industry. The large-scale commercial production of indigo relies on its extraction from plants and chemical synthesis. Studies are being conducted to develop methods for environment-friendly and sustainable production of indigo using genetically engineered microbes. Here, to enhance the yield of bioindigo from an E. coli whole-cell system containing tryptophanase (TnaA) and flavin-containing monooxygenase (FMO), we evaluated tryptophan transporters to improve the transport of aromatic compounds, such as indole and tryptophan, which are not easily soluble and passable through cell walls. Among the three transporters, Mtr, AroP, and TnaB, AroP enhanced indigo production the most. The combination of each transporter with AroP was also evaluated, and the combination of AroP and TnaB showed the best performance compared to the single transporters and two transporters. Bioindigo production was then optimized by examining the culture medium, temperature, isopropyl β-D-1-thiogalactopyranoside concentration, shaking speed (rpm), and pH. The novel strain containing aroP and tnaB plasmid with tnaA and FMO produced 8.77 mM (2.3 g/l) of bioindigo after 66 h of culture. The produced bioindigo was further recovered using a simple method and used as a watercolor dye, showing good mixing with other colors and color retention for a relatively long time. This study presents an effective strategy for enhancing indigo production using a combination of transporters.

Study on Intestinal Viability and Optimum Feeding Method of Lactobacillus in Broiler Chickens (육계에 대한 유산균의 장내 생존성 및 적정 급여방법에 대한 연구)

  • Kim, Dong-Wook;Kim, Ji-Hyuk;Kang, Geun-Ho;Kang, Hwan-Ku;Lee, Sang-Jin;Lee, Won-Jun;Kim, Sang-Ho
    • Journal of Animal Science and Technology
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    • v.50 no.6
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    • pp.807-818
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    • 2008
  • This study was conducted to prove the optimum feeding method of Lactobacillus in broiler chickens by investigating the intestinal viability of ingested Lactobacillus and the effect of feeding levels and frequency of Lactobacillus on growth performance in broiler chickens. In experiment 1, A total of one hundred, 5 weeks old male broiler chickens(Abor Acre) were fed Lactobacillus reuteri avibro2 expressed green fluorescent protein(GFP) at 104cfu/g diet to investigate the retention time of ingested Lactobacillus in the intestine for 1 day. The percentage of Lactobacillus expressed GFP in intestinal contents was 26% at 1 day after fed Lactobacillus expressed GFP. The percentage of Lactobacillus expressed GFP in intestinal contents was decreased in length of time. In experiment 2, A total of four hundred eighty, 1-d-old male broiler chicks(Abor Acre) were randomly divided into 4 groups with 4 replicates of 30 birds each to prove the optimum feeding level of Lactobacillus. The treatments were control(free antibiotics), Lactobacillus reuteri avibro2 5.0×10cfu/mL, 5.0×103cfu/mL, and 5.0×105cfu/mL. The final body weight and body wight gain of Lactobacillus reuteri avibro2 5.0×103cfu/mL were the highest in all groups(P<0.05). Feed conversion ratio was not significantly difference among the groups. The number of intestinal lactic acid bacteria in Lactobacillus treated groups tended to be improved or significantly increased as compared to that of control(P<0.05). Protein and fat digestibility in Lactobacillus 5.0×103cfu/mL and 5.0×105cfu/mL treated groups were significantly improved(P<0.05). No significant differences were observed on the availability of dry matter and crude ash in Lactobacillus treatments compared to those of control. In experiment 3, A total of six hundred 1-d-old male broiler chicks(Abor Acre) were randomly divided into 4 groups with 4 replicates of 30 birds each and were fed Lactobacillus reuteri avibro2 at intervals of 1, 2, 3, and 5 day for five weeks. Feeding level of Lactobacillus was 5.0×103cfu/mL The final body weight and body wight gain of Lactobacillus reuteri avibro2 5.0×103cfu/mL were the highest in all groups(P<0.05). The final body weight and body weight gain were significantly increased, when Lactobacillus was fed at intervals of 1 days, or 2 days. There were no significant differences in feed intake and feed conversion ratio among the all groups. The number of intestinal lactic acid bacteria in Lactobacillus treated groups tended to be improved or significantly increased as compared to that of control(P<0.05). No significant differences were observed on the number of coliform bacteria and Salmonella of ileum and cecum. Consequently, supplemental Lactobacillus influenced positive effects on the growth performance, nutrient availability and intestinal microflora. The optimum feeding level of Lactobacillus was 5.0×103cfu/mL, and the constant feeding of Lactobacillus was effective.

Biological treatment process for Food wastewater Using ER-1 bioreactor (다단 수직형 호기성반응기(ER-1)를 이용한 음식물 폐수의 생물학적 처리기술)

  • Lee, Jae-ki;Choi, Hong-Bok;Shin, Eung-Bai;Park, Ju-Hyoung;Choi, Eun-Ju;Kim, Jung-Rae;Park, Young-Sook
    • Journal of the Korea Organic Resources Recycling Association
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    • v.14 no.4
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    • pp.113-120
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    • 2006
  • Wastewater generated through the food waste recycling process have known high concentration, BOD 20,000~150,000 mg/L, which has to treat to the proper level because of a ban on reclamation. But it is impossible to treat less than 10 days by existing water treatment plant. Ecodays Ltd. is to treat this wastewater during 2~4 days by ER-1, which can simultaneously induce the modified PFR(Plug Flow Reactor) of the oxygen transfer rate, MLVSS concentration, and influent concentration to top from bottom of reactor. We tested the pilot test about low concentration food wastewater(BOD 16,500 mg/L) and high concentration food wastewater(64,431 mg/L) at the food waste recycling plant of H-Gun(20t/d). Hydraulic retention time(HRT) of ER-1 for low concentration food wastewater is 2.5day. In low concentration conditions, ER-1 treatment efficiency is to appear BOD 99%, COD 98%, TN 97%, and TP 96%. While ER-1 process for high concentration food wastewater treatment is composed 2 stages, which are to be HRT 2.5day for law wastewater and HRT 1.5 day for secondary treatment. In high concentration conditions, ER-1 treatment efficiency is to appear BOD 97%, COD 84%, TN 66%, and TP 95%. It is treated without temperature control about high temperature($50^{\circ}C$) to appear low treatment efficiency in high concentration conditions.

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Effect of Dietary Concentrate:forage Ratios and Undegraded Dietary Protein on Nitrogen Balance and Urinary Excretion of Purine Derivatives in Dorper×thin-tailed Han Crossbred Lambs

  • Ma, Tao;Deng, Kai-Dong;Tu, Yan;Jiang, Cheng-Gang;Zhang, Nai-Feng;Li, Yan-Ling;Si, Bing-Wen;Lou, Can;Diao, Qi-Yu
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.2
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    • pp.161-168
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    • 2014
  • This study aimed to investigate dietary concentrate:forage ratios (C:F) and undegraded dietary protein (UDP) on nitrogen balance and urinary excretion of purine derivatives (PD) in lambs. Four Dorper${\times}$thin-tailed Han crossbred castrated lambs with $62.3{\pm}1.9$ kg body weight at 10 months of age were randomly assigned to four dietary treatments in a $2{\times}2$ factorial arrangement of two levels of C:F (40:60 and 60:40) and two levels of UDP (35% and 50% of CP), according to a complete $4{\times}4$ Latin-square design. Each experimental period lasted for 19 d. After a 7-d adaptation period, lambs were moved into individual metabolism crates for 12 d including 7 d of adaption and 5 d of metabolism trial. During the metabolism trial, total urine was collected for 24 h and spot urine samples were also collected at different times. Urinary PD was measured using a colorimetric method and creatinine was measured using an automated analyzer. Intake of dry matter (DM) (p<0.01) and organic matter (OM) (p<0.01) increased as the level of UDP decreased. Fecal N was not affected by dietary treatment (p>0.05) while urinary N increased as the level of UDP decreased (p<0.05), but decreased as dietary C:F increased (p<0.05). Nitrogen retention increased as dietary C:F increased (p<0.05). As dietary C:F increased, urinary excretion of PD increased (p<0.05), but was not affected by dietary UDP (p>0.05) or interaction between dietary treatments (p>0.05). Daily excretion of creatinine was not affected by dietary treatments (p<0.05), with an average value of $0.334{\times}0.005$ mmol/kg $BW^{0.75}$. A linear correlation was found between total PD excretion and PDC index ($R^2$ = 0.93). Concentrations of creatinine and PDC index in spot urine were unaffected by sampling time (p>0.05) and a good correlation was found between the PDC index (average value of three times) of spot urine and daily excretion of PD ($R^2$ = 0.88). These results suggest that for animals fed ad libitum, the PDC index in spot urine is effective to predict daily excretion of PD. In order to improve the accuracy of the spot sampling technique, an appropriate lag phase between the time of feeding and sampling should be determined so that the sampling time can coincide with the peak concentration of PD in the urine.

Nutrient Recovery from Sludge Fermentation Effluent in Upflow Phosphate Crystallization Process (상향류 인 결정화공정을 이용한 슬러지 발효 유출수로 부터의 영양소 회수)

  • Ahn, Young-Ho
    • Journal of Korean Society of Environmental Engineers
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    • v.28 no.8
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    • pp.866-871
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    • 2006
  • The nutrient recovery in phosphate crystallization process was investigated by using laboratory scale uptlow reactors, adopting sequencing batch type configuration. The industrial waste lime was used as potential cation source with magnesium salt($MgCl_2$) as control. The research was focused on its successful application in a novel integrated sludge treatment process, which is comprised of a high performance fermenter followed by a crystallization reactor. In the struvite precipitation test using synthetic wastewater first, which has the similar characteristics with the real fermentation effluent, the considerable nutrient removal(about 60%) in both ammonia and phosphate was observed within $0.5{\sim}1$ hr of retention time. The results also revealed that a minor amount(<5%) of ammonia stripping naturally occurred due to the alkaline(pH 9) characteristic in feed substrate. Stripping of $CO_2$ by air did not increase the struvite precipitation rate but it led to increased ammonia removal. In the second experiment using the fermentation effluent, the optimal dosage of magnesium salt for struvite precipitation was 0.86 g Mg $g^{-1}$ P, similar to the mass ratio of the struvite. The optimal dosage of waste lime was 0.3 g $L^{-1}$, resulting in 80% of $NH_4-N$ and 41% of $PO_4-P$ removal, at about 3 hrs of retention time. In the microscopic analysis, amorphous crystals were mainly observed in the settled solids with waste lime but prism-like crystals were observed with magnesium salt. Based on mass balance analysis for an integrated sludge treatment process(fermenter followed by crystallization reactor) for full-scale application(treatment capacity Q=158,880 $m^3\;d^{-1}$), nutrient recycle loading from the crystallization reactor effluent to the main liquid stream would be significantly reduced(0.13 g N and 0.19 g P per $m^3$ of wastewater, respectively). The results of the experiment reveal therefore that the reuse of waste lime, already an industrial waste, in a nutrient recovery system has various advantages such as higher economical benefits and sustainable treatment of the industrial waste.

Study on the Thickness Effect of the Separator for Lithium Secondary Batteries (리튬이차전지용 분리막의 두께에 따른 특성 연구)

  • Kim, Sang Woo;Seok, Ji-Hoo;Kim, Byung-Hyun Daniel;Cho, Hee-Min;Cho, Kuk Young
    • Journal of the Korean Electrochemical Society
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    • v.17 no.1
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    • pp.7-12
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    • 2014
  • There is increasing demand on the reducing the weight and the volume of the major components in lithium secondary battery to improve energy density. Separator not only provides pathway for lithium ion movement but also prevents direct contact between anode and cathode. Herein we fabricated polyethylene separator by varying biaxial stretching ratio to obtain membrane thickness of 16, 12, and $9{\mu}m$. Mechanical and thermal properties of the separator with different thickness were investigated. Also rate capability and charge-discharge cycle property up to 500 cycles were studied using coin type full-cell with $LiCoO_2$ and graphite as a cathode and an anode, respectively. All the cells using separator with different thickness demonstrated excellent capacity retention after 500cycles (around 80%). Considering the rate capability, cell using separator with thickness of $9{\mu}m$ showed best performance. Interestingly, separator thickness of $9{\mu}m$ was more resistant to heat contraction compared to that of $16{\mu}m$ separator.

Behavior of Soluble Microbial Products in a Submerged Membrane Separation Activated Sludge Process (침지형 막분리 활성오니법에 있어서 생물대사성분의 거동)

  • Cha, Gi-Cheol;Lee, Dong-Yeol;Shim, Jin-Kie;Lee, Yong-Moo;Yoo, Ik-Keun;Ann, Seung-Ho
    • Journal of Korean Society of Environmental Engineers
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    • v.22 no.5
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    • pp.959-970
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    • 2000
  • A laboratory-scale experiment was conducted to investigate the effect of soluble microbial products(SMP) on permeate flux in the submerged membrane separation activated sludge process. Continuous and batch filtration test were operated to understand mechanism of relationship between membrane fouling and SMP. Synthetic wastewater(phenol) was used as a carbon source. Hydraulic retention time(HRT) and mixed-liquor volatile suspended solids(MLVSS) of the reactor were kept at 12 hours and 9.000mg VSS/L, respectively. Batch filtration tests ($J_{60}/J_o$) using the mixed liquor from reactor showed that the increase of accumulated SMP concentration in the reactor caused to the decreasing permeate flux and the increasing of the adhesion matters which form cake and gel layer. The resistance value of cake layer was measured $2.9{\times}10^{10}{\sim}4.0{\times}10^{10}(1/m)$, this value showed more significant effect on flux drop than that of among other resistance layers. Batch phenol-degradation experiment was conducted to observe SMP type expected $SMP_{nd}$ and $SMP_{e}$ (SMP resulted from endogenous cell decomposition), these are non-biodegradable high molecular weight organic matter and playa significant role in permeate flux drop. Also, SMP concentration was accumulated as increased of HRT against flux drop.

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Enhanced and Targeted Expression of Fungal Phytase in Saccharomyces cerevisiae

  • LIM, YOUNG-YI;EUN-HA PARK;JI-HYE KIM;SEUNG-MOON PARK;HYO-SANG JANG;YOUN-JE PARK;SEWANG YOON;MOON-SIK YANG;DAE-HYUK KIM
    • Journal of Microbiology and Biotechnology
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    • v.11 no.6
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    • pp.915-921
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    • 2001
  • Phytase improves the bioavailability of phytate phosphorus in plant foods to humans and animals, and reduces the phosphorus pollution of animal waste. In order to express a high level of fungal phytase in Saccharomyces cerevisiae, various expression vectors were constructed with different combinations of promoters, translation enhancers, signal peptides, and terminator. Three different promoters fused to the phytase gene (phyA) from Aspergillus niger were tested: a galactokinase (GAL1) promoter, glyceraldehyde-3-phosphate dehydrogenase (GPD) promoter, and yeast hybrid ADH2-GPD promoter consisting of alcohol dehydrogenase II (ADH2) and a GPD promoter. The signal peptides of phytase, glucose oxidase (GO), and rice amylase 1A(RAmy1A) were included. Plus, the translation enhancers of the ${\Omega}$ sequence and UTR70 from the tobacco mosaic virus (TMV) and spinach, respectively, were also tested. Among the recombinant vectors, pGphyA06 containing the GPD promoter, the ${\Omega}$ sequence, RAmy1A, and GAL7 terminator expressed the highest phytase activity in a culture filtrate, which was estimated at 20 IU/ml. An intracellular localization of the expressed phytase activity in a culture filtrate, which was estimated at 20 IU/ml. An intracellular localization of the expressed phytase was also performed by inserting an endoplasmic reticulum (ER) retention signal, KDEL sequence, into the C-terminus of the phytase within the vector pHphyA-6. It appeared that the KDEL sequence directed most of the early expression of phytase into the intracellular compartment yet more than $60\%$ of the total phytase activity was still retained within the cell even after the prolonged (>3 days) incubation of the transformant. However, the intracellular enzyme activity of the transformant without a KDEL sequence was as high as that of the extracellular one, thereby strongly suggesting that the secretion of phytase in S. cerevisiae appeared to be the rate-limiting step for the expression of a large amount of extracellular recombinant phytase, when compared with other yeasts.

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Biodegradability of Artificial Bait for Blue Crab Pots and Its Effect on Seawater Quality (꽃게 통발용 인공미끼의 생분해도 및 해양수질 영향)

  • Jeong, Byung-Gon;Koo, Jae-Geun;Chang, Ho-Young
    • Journal of the Korean Society for Marine Environment & Energy
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    • v.12 no.2
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    • pp.96-103
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    • 2009
  • The biodegradability in water of the artificial baits for blue crab pots which were made of intestines of mackerel, tuna and grinded krill were studied. The biodegradability of artificial bait was evaluated with the effective capacity of 10L water tank which was made of acryl pipe at the velocity of 1m/d and hydraulic retention time of 12 hours. For the 23 days operation time, all artificial baits were degraded fast at the early stage of operation time and stabilized within 5 days after start up. The rates of biodegradation were different depending on the raw materials of artificial baits. In terms of degradation rate of organic matter which can be expressed as COD, artificial bait made of tunas intestine showed the fastest degradation rate. On the other hand, in terms of degradation rate of nitrogenous matter which can be expressed as ammonia nitrogen, artificial bait made of mackerels intestine showed the fastest degradation rate. In order to evaluate the effect of artificial bait on marine ecosystem, seawater qualities including SS, COD, DO, nitrogen, phosphorus were determined depending on depth and location during 2 days test operation period. It is apparent that the effect of artificial bait on seawater quality was negligible when comparing seawater quality of test operation area with control area.

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Effect of the Addition of Granular Activated Carbon and Granular Sludge on the Performance of Upflow Anaerobic Sludge Blanket Reactors for Treating Leachate (상향류 혐기성 슬러지 블랭킷 반응조를 이용한 침출수 처리시 입상 활성탄 및 입상슬러지 첨가의 영향)

  • Lee, Chae-Young
    • Journal of the Korea Organic Resources Recycling Association
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    • v.16 no.4
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    • pp.91-97
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    • 2008
  • The objective of this research is to investigate the effect of the addition of granular activated carbon (GAC) and granular sludge on the performance of upflow anaerobic sludge blanket (UASB) reactors for treating leachate. For the control reactor, sludge obtained from an anaerobic digester was used as a seed material. On the other hand, GAC and granular sludge were incorporated with the seed sludge in the GAC reactor and the Granule reactor, respectively. The shortest acclimation period was observed in the Granule reactor. The GAC reactor also gave comparable performance to the Granule reactor at the beginning of operation. However, as the adsorptive capacity of GAC was exhausted, the effluent COD concentration increased gradually. Once the systems were stabilized, the GAC reactor showed slightly better results than the other two reactors in terms of chemical oxygen demand (COD) removal. COD removal in all reactors was more than 90% at hydraulic retention time of 1.0 day. Furthermore, GAC reactor showed little variation in COD removal rate and remained at 95% with organic loading rate (OLR) of 4.0 to $8.2kg\;COD/m^3.d$. Initial operating period was reduced by the addition of granular sludge, while the treatment efficiency was enhanced by the addition of GAC.

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