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        Oxygen-dependent enhancement of hydrogen production by engineering bacterial hemoglobin in Escherichia coli

        Jo, B.H.,Kim, J.Y.H.,Seo, J.H.,Cha, H.J. Pergamon Press ; Elsevier Science Ltd 2014 International journal of hydrogen energy Vol.39 No.20

        H<SUB>2</SUB> production under aerobic conditions has been proposed as an alternative method to overcome the fundamentally low yield of H<SUB>2</SUB> production by fermentative bacteria by maximizing the number of electrons that are available for H<SUB>2</SUB>. Here, we engineered Vitreoscilla hemoglobin (VHb) in Escherichia coli to study the effects of this versatile oxygen (O<SUB>2</SUB>)-binding protein on oxic H<SUB>2</SUB> production in a closed batch system that was supplemented with glucose. The H<SUB>2</SUB> yields that were obtained with the VHb-expressing E. coli were greatly enhanced in comparison to the negative control cells in culture that started with high O<SUB>2</SUB> tensions. The formate hydrogen lyase (FHL) activity of oxically cultured, VHb-expressing cells was also much higher than that of the negative control cells. Through inhibitor studies and time-course experiments, VHb was shown to contribute to the improved H<SUB>2</SUB> yield primarily by increasing the efficiency of cellular metabolism during the aerobic phase before the onset of H<SUB>2</SUB> production and not by working as an O<SUB>2</SUB>-scavenger during H<SUB>2</SUB> production. This new approach allowed more substrate to remain to be further utilized for the production of more H<SUB>2</SUB> from limited resources. We expect that VHb can be successfully engineered in potential aerobic H<SUB>2</SUB>-producing microbial systems to enhance the overall H<SUB>2</SUB> production yield. In addition, the remarkably high FHL activity of oxically grown, VHb-expressing cells may make this engineered strain an attractive whole-cell biocatalyst for converting formate to H<SUB>2</SUB>.


        A multi-virus detectable microfluidic electrochemical immunosensor for simultaneous detection of H1N1, H5N1, and H7N9 virus using ZnO nanorods for sensitivity enhancement

        Han, J.H.,Lee, D.,Chew, C.H.C.,Kim, T.,Pak, J.J. Elsevier Sequoia 2016 Sensors and actuators. B Chemical Vol.228 No.-

        This paper describes a multi-detectable and nano-flow immunosensor based on ZnO nanorods (NRs) grown on the inner surface of PDMS sensor region for sensing H1N1, H5N1, and H7N9 influenza viruses simultaneously using electrochemical method. Nanostructured ZnO NRs with a high isoelectric point (IEP ~9.5) tend to interact electrostatically with proteins with lower IEP such as H1N1, H5N1, and H7N9 antibodies. ZnO NRs were hydrothermally grown on the upper inner surface of the nano-flow PDMS sensor region. The forementioned three influenza viruses were successfully detected from three separate sensing regions by measuring the oxidation current of 3,3',5,5'-tetramethylbenzidine (TMB) by horseradish peroxidase (HRP) conjugated on capture antibody of those influenza viruses when proper potential was applied. The proposed immunosensors were evaluated using 1pg/ml, 10pg/ml, 100pg/ml, 1ng/ml, and 10ng/ml of H1N1, H5N1, and H7N9 antigens by amperometry. These immunosensors showed high selectivity toward H1N1, H5N1, and H7N9, which was successfully confirmed by distinguishing the target virus individually from a mixture of three virus antigens. A low limit of detection was demonstrated by detecting as low as 1pg/ml of each virus and it is believed that this was possible by enhancing the sensitivity with the ZnO NRs grown on the PDMS surface in the sensing region. The steady-state oxidation current output linearly increased with respect to the logarithm of the H1N1, H5N1, and H7N9 virus concentrations in the range of 1-10ng/ml.


        Experimental infection of mandarin duck with highly pathogenic avian influenza A (H5N8 and H5N1) viruses

        Kang, H.M.,Lee, E.K.,Song, B.M.,Heo, G.B.,Jung, J.,Jang, I.,Bae, Y.C.,Jung, S.C.,Lee, Y.J. Elsevier Scientific Pub. Co 2017 Veterinary microbiology Vol.198 No.-

        <P>A highly pathogenic avian influenza (HPAI) H5N8 virus was first detected in poultry and wild birds in South Korea in January 2014. Here, we determined the pathogenicity and transmissibility of three different clades of 1-15 viruses in mandarin ducks to examine the potential for wild bird infection. H5N8 (Glade replicated more efficiently in the upper and lower respiratory tract of mandarin ducks than two previously identified H5N1 virus clades (clades 2.2 and However, none of the mandarin ducks infected with H5N8 and H5N1 viruses showed severe clinical signs or mortality, and gross lesions were only observed in a few tissues. Viral replication and shedding were greater in H5N8-infected ducks than in H5N1-infected ducks. Recovery of all viruses from control duck in contact with infected ducks indicated that the highly pathogenic H5 viruses spread horizontally through contact. Taken together, these results suggest that H5N8 viruses spread efficiently in mandarin ducks. Further studies of pathogenicity in wild birds are required to examine possible long-distance dissemination via migration routes. (C) 2016 Elsevier B.V. All rights reserved.</P>

      • KCI등재

        돼지 H-FABP 유전자의 다형성 및 경제 형질과의 연관성 구명

        최봉환,김태헌,이지웅,조용민,이혜영,조병욱,정일정 한국동물자원과학회 2003 한국축산학회지 Vol.45 No.5

        The purpose of this study was to detect association between genetic variation and economic trait in the porcine heart type fatty acid-binding protein gene as a candidate gene for the traits related with growth and meat quality in pigs. The H-FABP is a 15-kDa protein expressed in several tissues with high demand for fat metabolism such as cardiac and skeletal muscle and lactating mammary gland. H-FABP is small intracellular protein involved in fatty acid transport from the plasma membrane to the site of β-oxidation and/or triacylglycerol or phospholipid synthesis. In this study, H-FABP PCR-RFLP was performed in F_(2) population composed of 214 individuals form an intercross between Korean Native Boars and Landrace sows. PCR products form tow primer sets within H-FABP gene were amplified in 850bp and 700bp. Digestion of PCR products with the restriction digestion enzymes HaeⅢ and Hinf Ⅰ, revealed fragment length polymorphisms(RFL. Ps). The genotype frequencies from H-FABP/HaeⅢ was .29 for genotype DD, .53 for genotype Dd, and .15 for genotype dd, respectively. The genotype frequencies of HH, Hh, and hh from H-FABP(hinf Ⅰ was .38, .41, and .20, respectively, in the population.Relationships between their genotypes and economic traits were estimated. In H-FABP/HaeⅢ locus, there were specific genotypes(Dd and dd) associated with economic traits such as body weight. In H-FABP/Hinf Ⅰ Iocus, Genotypes of HH and Hh associated with growth traits such as body weights at 5, 12, and 30 week of age (p<.05 or p<.001) and back fat thickness, body fat including abdominal and trimmed fat (p<.001) and intramuscular fat(p<.05). The 'H'allele was positivecly associated with gaining of body weight and fatness deposition. In conclusion, a significant association of the H-FABP gene from its genetic variation was found on body weight, intramuscular fat and backfat thickness.

      • Expression and N-glycan analysis of human 90K glycoprotein in Drosophila S2 cells

        Kim, K.R.,Kim, Y.K.,Cheong, H.,Kim, J.Y.H.,Cha, H.J. IPC Science and Technology Press ; Elsevier Scienc 2013 Enzyme and microbial technology Vol.53 No.3

        Human 90K (h90K; Mac-2-binding protein) glycoprotein is a potential pharmaceutical due to its inhibitory activity against cancer metastasis and expansion. Here, h90K glycoprotein was produced in insect Drosophila S2 cell system, and its N-glycan pattern was analyzed. A plasmid encoding h90K gene, fused with a hexahistidine tag under the control of Drosophila metallotionein promoter, was stably transfected into S2 cells. After copper sulfate induction, transfected S2 cells secreted recombinant h90K at a good expression level of 28mg/L in a 150-mL spinner flask culture. The purified recombinant h90K showed an apparent molecular weight of ~78kDa which was much smaller than that (~97kDa) of the natural h90K. Because de-N-glycosylated h90K appeared at ~60kDa protein band, it was suggested that the recombinant h90K from S2 cells has small N-glycans with about half the molecular weight (~18kDa) of N-glycans of the natural h90K. Through detail analyses using high-performance liquid chromatography and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, the S2-derived recombinant h90K was confirmed that it has simple paucimannosidic structures containing two or three mannose residues with core fucose as the major (~79%) N-glycans.

      • KCI등재

        Selection for Duration of Fertility and Mule Duck White Plumage Colour in a Synthetic Strain of Ducks (Anas platyrhynchos)

        H.C. Liu,J.F. Huang,S.R. Lee,H.L. Liu,C.H. Hsieh,C.W. Huang,M.C. Huang,C. Tai,J.P. Poivey,R. Rouvier,Y.S. Cheng 아세아·태평양축산학회 2015 Animal Bioscience Vol.28 No.5

        A synthetic strain of ducks (Anas platyrhynchos) was developed by introducing genes for long duration of fertility to be used as mother of mule ducklings and a seven-generation selection experiment was conducted to increase the number of fertile eggs after a single artificial insemination (AI) with pooled Muscovy semen. Reciprocal crossbreeding between Brown Tsaiya LRI-2 (with long duration of fertility) and Pekin L-201 (with white plumage mule ducklings) ducks produced the G0. Then G1 were intercrossed to produce G2 and so on for the following generations. Each female duck was inseminated 3 times, at 26, 29, and 32 weeks of age. The eggs were collected for 14 days from day 2 after AI. Individual data regarding the number of incubated eggs (Ie), the number of fertile eggs at candling at day 7 of incubation (F), the total number of dead embryos (M), the maximum duration of fertility (Dm) and the number of hatched mule ducklings (H) with plumage colour were recorded. The selection criterion was the breeding values of the best linear unbiased prediction animal model for F. The results show high percentage of exhibited heterosis in G2 for traits to improve (19.1% for F and 12.9% for H); F with a value of 5.92 (vs 3.74 in the Pekin L-201) was improved in the G2. Heritabilities were found to be low for Ie (h2 = 0.07±0.03) and M (h2 = 0.07±0.01), moderately low for Dm (h2 = 0.13±0.02), of medium values for H (h2 = 0.20±0.03) and F (h2 = 0.23±0.03). High and favourable genetic correlations existed between F and Dm (rg = 0.93), between F and H (rg = 0.97) and between Dm and H (rg = 0.90). The selection experiment showed a positive trend for phenotypic values of F (6.38 fertile eggs in G10 of synthetic strain vs 5.59 eggs in G4, and 3.74 eggs in Pekin L-201), with correlated response for increasing H (5.73 ducklings in G10 vs 4.86 in G4, and 3.09 ducklings in Pekin L-201) and maximum duration of the fertile period without increasing the embryo mortality rate. The average predicted genetic response for F was 40% of genetic standard deviation per generation of selection. The mule ducklings’ feather colour also was improved. It was concluded that this study provided results for a better understanding of the genetics of the duration of fertility traits in the common female duck bred for mule and that the selection of a synthetic strain was effective method of improvement.

      • KCI등재후보

        Effects of Taurine on Sperm Characteristics during In vitro Storage of Boar Semen

        H.Y. Jang,H.S. Kong,박춘근,J.D. Oh,S.G. Lee,정희태,김종택,이성진,양부근,이학교 아세아·태평양축산학회 2006 Animal Bioscience Vol.19 No.11

        The objective of this study was to investigate the anti-oxidative effects of taurine on sperm characteristics for in vitro storage of boar semen. Semen was randomly divided into 10 groups in conical tubes and treated with different concentrations of taurine (25-100 mM) with or without 250 ???H2O2. The percentage of motile spermatozoa in taurine groups after 6 and 9 h were significantly higher at >94% and 87%, respectively, compared to the control group (85.1??.5 and 72.4??.3, p<0.05). The sperm motility in taurine with H2O2 after 6 h incubation was slightly decreased compared to the taurine alone treatment, but after 9 and 12 h incubation % sperm motility dropped sharply in taurine with H2O2 (75.3??.3 and 69.6??.9, p<0.05). For 3, 9 and 12 h incubation, sperm viability in the control was lower than in taurine groups, irrespective of taurine concentration. In eosin Y and nigrosin staining (ENS), the sperm survival rates (%) for 6 h incubation were significantly higher in 25 mM (76.0??.6) and 50 mM taurine groups (78.0??.7), respectively. Sperm survival rates for 9 and 12 h incubation were higher in taurine groups (??8% in 9 h and ??2% in 12 h) compared to controls (43.0??.1 and 31.0??.6, respectively). In the hyoosmotic swelling test (HOST), sperm membrane integrity was similar to the results of sperm survival. These experiments indicate that supplementation of taurine to the semen extender can increase the sperm characteristics (motility, viability, survival and membrane integrity).

      • KCI등재

        A comparative study on the performance of different advanced oxidation processes (UV/O3/H2O2) treating linear alkyl benzene (LAB) production plant's wastewater

        H. Zangeneh,A.A.L. Zinatizadeh,M. Feizy 한국공업화학회 2014 Journal of Industrial and Engineering Chemistry Vol.20 No.4

        A detailed investigation on photooxidation of linear alkyl benzene (LAB) industrial wastewater is presented in this study. The process analysis was performed by varying four significant independent variables including two numerical factors (initial pH (3–11) and initial H2O2 concentration (0–20 mM)) and two categorical factors (UV irradiation and ozonation). The experiments were conducted based on a central composite design (CCD) and analyzed using response surface methodology (RSM). To assess the process performance, two parameters viz. TCOD removal efficiency and BOD5/COD were measured throughout the experiments. A maximum reduction in TCOD was 58, 53, 51, and 49%, respectively for UV/H2O2/O3, H2O2/O3, UV/O3 and UV/H2O2 processes at the optimum conditions (initial pH of 7, initial H2O2 concentration of 100 mM, and reaction time of 180 min). A considerable increase in BOD5/COD ratio was obtained in the combined processes (0.46, 0.51, 0.53, and 0.55 for UV/H2O2, UV/O3, H2O2/O3 and UV/H2O2/O3, respectively) compared to the single oxidant process (0.35). The results showed that mineralization of the LAB industrial wastewater in neutral pH is more favored than in acidic and basic pH. Gas chromatography–mass spectrometry (GC–MS) was applied to show the fate of organic compounds. In conclusion, the photooxidation process (UV/H2O2/O3, H2O2/O3, UV/O3 and UV/H2O2) could be an appropriate pretreatment method prior to a biological treatment process.


        On the zeolitic imidazolate framework-8 (ZIF-8) membrane for hydrogen separation from simulated biomass-derived syngas

        Yin, H.,Lee, T.,Choi, J.,Yip, A.C.K. Elsevier 2016 Microporous and mesoporous materials Vol.233 No.-

        <P>Hydrogen separation from biomass-derived syngas is a critical step in the utilization of gasification technology. Compared with the traditional methods, membrane technology provides an effective and low-cost solution for adjusting the gas composition and collecting H-2 in syngas environments. In this study, a zeolitic imidazolate framework-8 (ZIF-8)-based membrane, which is a potential candidate for H-2 separation from biomass-derived syngas, was successfully fabricated through the seeded (secondary) growth method and the subsequent post-treatments. The prepared ZIF-8 membrane exhibited a modest H-2 separation performance for H-2/CO2 and H-2/CO, with separation factors of 4.95 and 6.08, respectively, and a H-2 permeance of 7.81 x 10(-8) mol m(-2) s(-1) Pa-1 at 200 degrees C in the simulated biomass-derived syngas environments (H-2/CO2/CO) with the presence of steam. In particular, the H-2/CO2 and H-2/CO separation factors were increased by 36% and 97%, respectively, with respect to those obtained through bare supports. Despite the promising H2 perm-selectivity, the H-2/CO2 and H-2/CO separation factors of ZIF-8 membranes at 200 degrees C under water-containing syngas environments were maintained up to 10 h but the longer exposure led to the gradual degradation and eventual reduction toward those of bare supports after 15 h seemingly due to the water-involved membrane degradation. This study provides the availability and limitation of ZIF-8 membranes for H-2 separations in stimulated biomass-derived syngas environments. (C) 2015 Elsevier Inc. All rights reserved.</P>


        Surveillance of avian influenza virus in wild bird fecal samples from South Korea, 2003-2008.

        Kang, H M,Jeong, O M,Kim, M C,Kwon, J S,Paek, M R,Choi, J G,Lee, E K,Kim, Y J,Kwon, J H,Lee, Y J [Wildlife Disease Association] 2010 JOURNAL OF WILDLIFE DISEASES Vol.46 No.3

        <P>We analyzed the results from nationwide surveillance of avian influenza (AI) from birds in South Korea's major wild bird habitats and the demilitarized zone of South Korea, 2003-2008. Of 28,214 fecal samples analyzed, 225 yielded influenza viruses, for a prevalence of 0.8%. Hemagglutinin (HA) subtypes H1-H12 and all nine neuraminidase (NA) subtypes were detected. The dominant HA subtypes were H6, H1, and H4, and the most common NA subtypes were N2, N1, and N6. Among the 38 HA/NA subtype combinations, the most common were H4N6, H6N1, and H5N2. Thirty-seven low-pathogenic AI (LPAI) viruses of the H5 and H7 subtype were detected. Among them, we identified bird species for 16 H5- and H7-positive fecal samples using a DNA bar-coding system instituted in 2007; all birds were identified as Anseriformes. The HA gene of the H5 wild bird isolates belonged to the Eurasian avian lineage, and could be clearly distinguished from the sublineage H5N1 highly pathogenic AI (HPAI) of the Eurasian and American avian lineages. Whereas H7 LPAI viruses did not group as a separate sublineage with H7 HPAI viruses, H7 isolates were closely related with the Eurasian avian lineage.</P>

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