• Title/Summary/Keyword: Glycerol analysis

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Bacterial Expression of the scFv Fragment of a Recombinant Antibody Specific for Burkholderia pseudomallei Exotoxin

  • Su, Yu-Ching;Lim, Kue-Peng;Nathan, Sheila
    • BMB Reports
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    • v.36 no.5
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    • pp.493-498
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    • 2003
  • The scFv antibody towards the Burkholderia pseudomallei exotoxin was previously constructed by phage display and exhibited good specificity towards the exotoxin. We report here the optimization of the scFv expression in an E. coli expression system. Four different E. coli strains (ER2537, TG1, HB2151, and XL1-Blue) were examined for optimal expression of the scFv protein. Two types of carbon source (i.e. 0.2% glucose and 0.2% glycerol) were also tested for their ability to induce the scFv expression. Cells that carried the scFv construct were grown at $30^{\circ}C$ and induced with 0.05 mM IPTG. The expression was then monitored by SDS-PAGE, Western blotting, and indirect ELISA. The Western blot profile showed different levels of the scFv expression among the host strains; XL1-Blue exhibited the highest level of the scFv protein expression. Glycerol at a concentration of 0.2% (v/v) significantly increased the scFv protein expression level when compared to 0.2% (w/v) glucose. Further optimization demonstrated that the scFv protein expression in XL1-Blue was the most optimal with a glycerol concentration as low as 0.05%. However, by indirect ELISA, only the scFv protein that was expressed in 0.2% (v/v) glycerol exhibited high specificity towards the Burkholderia pseudomallei exotoxin.

A Study for the Removal of Phosphorous Using Coated Exfoliated Vermiculite (인 제거를 위한 코팅 발포질석 적용 가능성 연구)

  • Kim, Seogku;Lee, Taeyoon
    • Journal of the Korean GEO-environmental Society
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    • v.15 no.12
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    • pp.5-13
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    • 2014
  • In this study, exfoliated vermiculite (EV) coated with glycerol was tested for its abiility to remove phosphorus in aqueous solution. The glycerol modified vermiculite (GS) was prepared with EV/glycerol ratio of 1/4 where glycerol contained 4 mol% $H_2SO_4$ and heated until designated temperature. GS heated at $380^{\circ}C$ showed that the specific surface area was $53.1m^2/g$ and mass loss due to oxidation of carbon was maximum from TGA analysis. Removal of phosphorus using GS heated at $380^{\circ}C$ was well explained by Langmuir isotherm model and maximum sorption capacity of 714.3 mg/kg is comparable or greater than those of other clay orignated sorbents for phosphorus.

Influence of Osmolality of Complete Semen Extender on Motion Characteristics of Frozen-thawed Ram Spermatozoa

  • Joshi, Anil;Mathur, A.K.;Naqvi, S.M.K.;Mittal, J.P.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.12
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    • pp.1716-1721
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    • 2006
  • The present study was conducted to observe the effect of osmolality of glycerolated TEST-yolk glycerol extenders on post-thawing sperm kinematics of ram spermatozoa of the native Malpura breed maintained in a semi-arid tropical environment. Good quality semen obtained from adult rams was pooled, split and diluted to 1,000 million spermatozoa per ml in complete TEST-yolk-glycerol extenders of 900, 1,200, 1,500 and 1,800 mOsm/kg osmolality. Diluted semen samples were loaded in 0.25 ml straws and cooled down to $-125^{\circ}C$ freezing temperature at the rate of $-25^{\circ}C$ per minute under controlled conditions before plunging into liquid nitrogen for storage. The thawing of straws was performed at $50^{\circ}C$ in a water bath for 10 seconds and sperm kinematics of the frozen-thawed spermatozoa were assessed by a computer-assisted sperm analysis technique. Osmolality of diluent had no significant effect on post-thawing % motility, % rapid, % medium and % slow moving frozen-thawed spermatozoa but significantly (p< 0.05) affected the % linearity and % straightness. The post-thawing % motility and % rapid motile spermatozoa were highest in samples extended in diluent of 1,500 mOsm/kg osmolality and lowest in 900 mOsm/kg. The curvilinear velocity of spermatozoa was significantly (p<0.05) higher for samples extended in 1,800 mOsm/kg, compared to those in 900 and 1,200 mOsm/kg, but the effect was not significantly different to those extended in diluent of 1,500 mOsm/kg osmolality. The study indicated that ram spermatozoa could tolerate a wide osmolality range for dilution in the complete TEST-yolk-glycerol extender for their cryosurvival. The highest recovery of motile spermatozoa following thawing was achieved in samples extended in the TEST-yolk-glycerol diluent of 1,500 mOsm/kg osmolality.

Statistical Optimization of Medium Components by Response Surface Methodology to Enhance Menaquinone-7 (Vitamin K2) Production by Bacillus subtilis

  • Wu, Wei-Jie;Ahn, Byung-Yong
    • Journal of Microbiology and Biotechnology
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    • v.28 no.6
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    • pp.902-908
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    • 2018
  • Optimization of the culture medium to maximize menaquinone-7 (MK-7) production by Bacillus subtilis strain KCTC 12392BP in static culture was carried out using statistical experimental methods, including one factor at a time, fractional factorial design, and response surface methodology (RSM). Maltose (carbon source), tryptone (nitrogen source), and glycerol (activator) were identified as the key medium components for MK-7 synthesis by the fractional factorial design, and were selected for statistical optimization by RSM. The statistical analysis indicated that, in the range that was studied, maltose, tryptone, and glycerol were all critical factors having profound effects on the production of MK-7, with their coefficients for linear and quadratic all significant at the p < 0.05 level. The established model was efficient and feasible, with a determination coefficient ($R^2$) of 0.9419. The predicted concentrations of maltose, tryptone, and glycerol in the optimal medium were determined as 36.78, 62.76, and 58.90 g/l, respectively. In this optimized medium, the maximum yield of MK-7 reached a remarkably high level of $71.95{\pm}1.00{\mu}g/ml$ after 9 days of static fermentation, which further verified the practicability of this optimized strategy.

The biomechanical and biological effect of supercooling on cortical bone allograft

  • MuYoung Kim ;Hun-Young Yoon
    • Journal of Veterinary Science
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    • v.24 no.6
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    • pp.79.1-79.16
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    • 2023
  • Background: The need for a storage method capable of preserving the intrinsic properties of bones without using toxic substances has always been raised. Supercooling is a relatively recently introduced preservation method that meets this need. Supercooling refers to the phenomenon of liquid in which the temperature drops below its freezing point without solidifying or crystallizing. Objectives: The purpose of this study was to identify the preservation efficiency and applicability of the supercooling technique as a cortical bone allograft storage modality. Methods: The biomechanical effects of various storage methods, including deep freezing, cryopreservation, lyophilization, glycerol preservation, and supercooling, were evaluated with the three-point banding test, axial compression test, and electron microscopy. Additionally, cortical bone allografts were applied to the radial bone defect in New Zealand White rabbits to determine the biological effects. The degree of bone union was assessed with postoperative clinical signs, radiography, micro-computed tomography, and biomechanical analysis. Results: The biomechanical properties of cortical bone grafts preserved using glycerol and supercooling method were found to be comparable to those of normal bone while also significantly stronger than deep-frozen, cryopreserved, and lyophilized bone grafts. Preclinical research performed in rabbit radial defect models revealed that supercooled and glycerol-preserved bone allografts exhibited significantly better bone union than other groups. Conclusions: Considering the biomechanical and biological superiority, the supercooling technique could be one of the optimal preservation methods for cortical bone allografts. This study will form the basis for a novel application of supercooling as a bone material preservation technique.

Evaluation of Newly Isolated Klebsiella pneumoniae Strains for the Co-Production of 3-hydroxypropionic acid and 1,3-propanediol from Glycerol (새로이 분리된 Klebsiella pneumoniae 균주들의 글리세롤 기반 3-hydroxypropionic acid 및 1,3-propanediol 동시 생산성 평가)

  • Ko, Yeounjoo;Seol, Eunhee;Sekar, Balaji Sundara;Kwon, Seongjin;Lee, Jaehyeon;Park, Sunghoon
    • KSBB Journal
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    • v.31 no.4
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    • pp.246-255
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    • 2016
  • Co-production of 3-hydroxypropionic acid (3-HP) and 1,3-propanediol (1,3-PDO) was suggested as an innovative strategy to overcome several limitations occurring in the single production of 3-HP from glycerol. In this study, two new isolates of Klebsiella pneumoniae, which produce less lipopolysaccharide (LPS) thus considered less pathogenic than K. pneumoniae DSM 2026, were compared and evaluated for their potential for the co-production of 3-HP and 1,3-PDO. The newly isolated strains showed significantly faster sedimentation rate than DSM, which should be beneficial for downstream processing. Analysis of genome sequences of the isolates confirmed the presence of all genes necessary for glycerol assimilation, 1,3-PDO production and biosynthesis of coenzyme $B_{12}$. Co-production yield was highest under anaerobic condition while cell growth was highest under aerobic condition. Both strains showed similarly good performance for the co-production although J2B gave the slightly higher co-production yield of 0.80 mol/mol than GSC021 (0.75 mol/mol). The evaluation of the newly developed strains presented here should be useful in designing similar evaluation experiments for other microorganisms.

Development of reference materials for cement paste

  • Lee, Dong Kyu;Choi, Myoung Sung
    • Advances in concrete construction
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    • v.9 no.6
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    • pp.547-556
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    • 2020
  • This study aimed to develop reference materials (RMs) that are chemically stable and can simulate the flow characteristics of cement paste. To this end, the candidate components of RMs were selected considering the currently required properties of RMs. Limestone, slag, silica, and kaolin were selected as substitutes for cement, while glycerol and corn syrup were selected as matrix fluids. Moreover, distilled water was used for mixing. To select the combinations of materials that meet all the required properties of RMs, flow characteristics were first analyzed. The results revealed that silica and kaolin exhibited bilateral nonlinearity. When an analysis was conducted over time, slag exhibited chemical reactions, including strength development. Moreover, fungi were observed in all mixtures with corn syrup. On the other hand, the combination of limestone, glycerol, and water exhibited a performance that met all the required properties of RMs. Thus, limestone, glycerol, and water were selected as the components of the RMs. When the influence of each component of the RMs on flow characteristics was analyzed, it was found that limestone affects the yield value, while the ratio of water and glycerol affects the plastic viscosity. Based on this, it was possible to select the mixing ratios for the RMs that can simulate the flow characteristics of cement paste under each mixing ratio. This relationship was established as an equation, which was verified under various mixing ratios. Finally, when the flow characteristics were analyzed under various temperature conditions, cement paste and the RMs exhibited similar tendencies in terms of flow characteristics. This indicated that the combinations of the selected materials could be used as RMs that can simulate the flow characteristics of cement paste with constant quality under various mixing ratio conditions and construction environment conditions.

Economic Evaluation with Uncertainty Analysis of Glycerol Steam Reforming for the H2 Production Capacity of 300 m3 h-1 (수소 생산 규모 300 m3 h-1급 글리세롤 수증기 개질반응에 대한 경제적 불확실성 분석)

  • Heo, Juheon;Lee, Boreum;Kim, Sehwa;Kang, Sung-Mook;Lim, Hankwon
    • Applied Chemistry for Engineering
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    • v.29 no.5
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    • pp.589-593
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    • 2018
  • In this paper, an economic evaluation with the uncertainty analysis using a Monte-Carlo simulation method was performed for the glycerol steam reforming to produce $H_2$ at a capacity of $300m^3h^{-1}$. Fluctuations in a unit $H_2$ production cost were identified based on the variation of key economic factors at ${\pm}10-{\pm}40%$ and the probability of 30.9% was obtained for a previously reported unit $H_2$ production cost of 5.10 $ $kgH{_2}^{-1}$. In addition, fluctuations in the B/C ratio were obtained by varying the fixed capital investment (${\pm}20%$), cost of manufacturing (${\pm}20%$), revenue (${\pm}20%$), and discount rate (2-10%) and the probability ranging from 17 to 55% was observed to meet a minimum B/C ratio of 1 for the economic feasibility of the glycerol steam reforming to produce $H_2$.

Visible Surface-Assisted Laser Desorption/Ionization Mass Spectrometry of Small Macromolecules Deposited on the Graphite Plate

  • Kim, Jung-Hwan;Paek, Kyung-Soo;Kang, Wee-Kyung
    • Bulletin of the Korean Chemical Society
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    • v.23 no.2
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    • pp.315-319
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    • 2002
  • Visible surface-assisted desorption/ionization mass spectrometry (SALDI-MS) has been investigated for several small macromolecules deposited on the graphite plate using laser radiation at 532 nm where most of the macromolecules are transparent. The graphite surface functioned well as a photon absorbing material and an energy transfer mediator for visible light. The results show that visible SALDI is a much softer ionization technique than UV-MALDI and FAB-MS in our results with synthetic macromolecules, PPG, PPGMBE and cavitand molecules. For the SALDI of biomolecules, glycerol as a proton source was essential with the graphite plate. As in visible SALDI, the role division of the photon absorbing material and the cationization agent can provide a generality in mass spectrometric analysis of macromolecules compared with MALDI using the dual functional matrix.

Cytotoxic Constituents of Diadema setosum

  • Minh, Chau-Van;Kiem, Phan-Van;Huong, Le-Mai;Kim, Young-Ho
    • Archives of Pharmacal Research
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    • v.27 no.7
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    • pp.734-737
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    • 2004
  • 5,8-Epidioxycholest-6-en-3-ol (1), cholesterol (2), glycerol 1-palmitate (3) and glycerol 1,3-dioleate-2-stearate (4) were isolated from the methanol extract of the sea urchin Diadema setosum, which was collected from the Halong sea, Vietnam. Chemical structures were established based on extensive 1D, 2D-NMR, FAB-MS, EI-MS spectroscopic data and GC-MS analysis. The NMR spectral data of compound 1 were reassigned by using HMQC and HMBC. Compound 1 was found to have strong cytotoxic effect against various cancer cell lines, such as KB ($IC_{50}$, 2.0 $\mu\textrm{g}$/mL), FL($IC_{50}$, 3.93 $\mu\textrm{g}$/mL), and Hep-2 ($IC_{50}$, 2.4 $\mu\textrm{g}$/mL) by in vitro assay.