• Applied Biological Chemistry
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• v.38 no.4
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• pp.320-324
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• 1995

Solubilization of microsomal ${\Delta}^{5}-3{\beta}$-hydroxy steroid acyl transfearse(${\Delta}^{5}-3{\beta}$-OH-SAT) of rat brain and its reconstitution into liposomes were investigated. Among the detergents utilized for the solubilization, deoxycholic acid was superior to Tween 80 or Triton X-100 for the reconstituted activity of ${\Delta}^{5}-3{\beta}$-OH-SAT. The enzyme activity was shown to be affected by the nature of phospholipids used for the preparation of the liposome. Phosphatidylcholines from egg yolk and soybean showed the highest activity of ${\Delta}^{5}-3{\beta}$-OH-SAT and phosphatidylethanolamine came next. However phosphatidylserine and phosphatidic acid showed a lower activity than those obtained before the reconstitution. This study suggests that the presence of quaternary ammonium salt or amine group in the phospholipids stimulates the activity of ${\Delta}^{5}-3{\beta}$-OH-SAT. However the presence of a carboxylic group or the absence of the amine group may have an inhibitory effect on the ${\Delta}^{5}-3{\beta}$-OH SAT.

• Journal of Soil and Groundwater Environment
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• v.11 no.5
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• pp.43-50
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• 2006

Bioreporter bacteria, such as recombinant bioluminescent bacteria, have been used for the detection of specific compounds in complex environmental media. In this study, optimum conditions for the preparation and application of deep-freezed and Iyophilized recombinant bioluminescent strain KG1206 were investigated for the future application on contaminated environmental sites. Genetically engineered microorganism, Pseudomonas putida mt-2 KG1206, contains TOL plasmid and the plasmid inserted $P_{m}$, promoter on the upper part of lux gone in vector pUCD615, and m-toluate and benzoate are considered direct inducers for bioluminescence. Optimum conditions determined for the preparation and application of the deep-freezed and lyophilized strain were followings: cryoprotective agent (24% sucrose), lyophilization time (12 hrs), strain concentration ($OD_{600}=0.6$), reconstitution for freezed strain (quick reconstitution at $35^{\circ}C$), reconstitution for lyophilized strain ($3{\sim}6$ hrs exposure on LB medium), carrying conditions (keep at $20^{\circ}C$ after reconstitution). These results demonstrate the feasibility of deep-freezed or lyophilized state of genetically engineered bioluminescent strain for environmental usage.

• Journal of Microbiology and Biotechnology
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• v.20 no.4
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• pp.727-731
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• 2010

The gene APE0743 encoding the superoxide dismutase (ApSOD) of a hyperthermophilic archaeon Aeropyrum pernix K1 was cloned and overexpressed as a GST fusion protein at a high level in Escherichia coli. The expressed protein was simply purified by the process of glutathione affinity chromatography and thrombin treatment. The ApSOD was a homodimer of 25 kDa subunits and a cambialistic SOD, which was active with either Fe(II) or Mn(II) as a cofactor. The ApSOD was highly stable against high temperature. This thermostable ApSOD is expected to be applicable as a useful biocatalyst for medicine and bioindustrial processes.

• BMB Reports
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• v.53 no.9
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• pp.466-471
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• 2020

Several humanized mouse models are being used to study humanspecific immune responses and diseases. However, the pivotal needs of fetal tissues for the humanized mice model have been huddled because of the demand for ethical and medical approval. Thus, we have verified the hematopoietic and immunomodulatory function of HepaRG and developed a new and easy humanized mouse model to replace the use of fetal liver tissue. HepaRG co-transplanted Hu-NSG mice significantly increased CD45+ lymphocytes and CD19+ B cells and CD3+ T cells than normal Hu-NSG, suggesting enhanced reconstitution of the human immune system. These results have improved the applicability of humanized mice by developing new models easily accessible.

• BMB Reports
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• v.35 no.3
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• pp.255-261
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• 2002

GTP cyclohydrolase I (E.C. 3.5.4.16) is a homodecameric protein that catalyzes the conversion of GTP to 7,8-dihydroneopterin triphosphate (H2NTP), the initial step in the biosynthesis of pteridines. It was proposed that the enzyme complex could be composed of a dimer of two pentamers, or a pentamer of tightly associated dimers; then the active site of the enzyme was located at the interface of three monomers (Nar et al. 1995a, b). Using mutant enzymes that were made by site-directed mutagenesis, we showed that a decamer of GTP cyclohydrolase I should be composed of a pentamer of five dimers, and that the active site is located between dimers, as analyzed by a series of size exclusion chromatography and the reconstitution experiment. We also show that the residues Lys 136, Arg139, and Glu152 are of particular importance for the oligomerization of the enzyme complex from five dimers to a decamer.

• Food Science of Animal Resources
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• v.38 no.6
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• pp.1273-1285
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• 2018

The aim of this study is to determine the some physicochemical (proximate composition, pH and $a_w$ values) and reconstitution (wettability, dispersibility and solubility index (SI)) properties of spray-dried bone broth powder (BBP) along with the effects of the addition of different carrier agents. Subsequently, the powdered products were stored to determine the storage stability (pH, lipid oxidation, color, browning index) for 3 mon at $-18^{\circ}C$. For this purpose, firstly marrow-containing bones (Os femur and Os humerus) have been boiled to get the bone broth. Three different emulsions were respectively prepared (1) 20% maltodextrin (MD) added BBP, (2) 20% whey powder isolate (WPI) added BBP and (3) 10% MD and 10% WPI added BBP and the emulsions were dried using a spray-dryer with $185^{\circ}C$ inlet and $95^{\circ}C$ outlet temperature. The proximate composition and reconstitution properties of BBP were found statistically different (p<0.05) depending on the use of different carrier agents. MD added BBP showed uniform and smooth morphology. The color, browning index, thiobarbituric acid reactive substances (TBARS) and pH values of BBP significantly changed (p<0.05) during storage. In conclusion, both the results of physicochemical and storage period analyses showed that the most suitable encapsulation material in the production of the bone broth powder is MD.

• BMB Reports
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• v.54 no.3
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• pp.157-163
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• 2021

The transient interactions between cellular components, particularly on membrane surfaces, are critical in the proper function of many biochemical reactions. For example, many signaling pathways involve dimerization, oligomerization, or other types of clustering of signaling proteins as a key step in the signaling cascade. However, it is often experimentally challenging to directly observe and characterize the molecular mechanisms such interactions-the greatest difficulty lies in the fact that living cells have an unknown number of background processes that may or may not participate in the molecular process of interest, and as a consequence, it is usually impossible to definitively correlate an observation to a well-defined cellular mechanism. One of the experimental methods that can quantitatively capture these interactions is through membrane reconstitution, whereby a lipid bilayer is fabricated to mimic the membrane environment, and the biological components of interest are systematically introduced, without unknown background processes. This configuration allows the extensive use of fluorescence techniques, particularly fluorescence fluctuation spectroscopy and single-molecule fluorescence microscopy. In this review, we describe how the equilibrium diffusion of two proteins, K-Ras4B and the PH domain of Bruton's tyrosine kinase (Btk), on fluid lipid membranes can be used to determine the kinetics of homodimerization reactions.

• Clinical and Experimental Pediatrics
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• v.56 no.1
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• pp.26-31
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• 2013

Purpose: Lymphocyte subset recovery is an important factor that determines the success of hematopoietic stem cell transplantation (HSCT). Temporal differences in the recovery of lymphocyte subsets and the factors influencing this recovery are important variables that affect a patient's posttransplant immune reconstitution, and therefore require investigation. Methods: The time taken to achieve lymphocyte subset recovery and the factors influencing this recovery were investigated in 59 children who had undergone HSCT at the Department of Pediatrics, The Catholic University of Korea Seoul St. Mary's Hospital, and who had an uneventful follow-up period of at least 1 year. Analyses were carried out at 3 and 12 months post-transplant. An additional study was performed 1 month post-transplant to evaluate natural killer (NK) cell recovery. The impact of pre- and post-transplant variables, including diagnosis of Epstein-Barr virus (EBV) DNAemia posttransplant, on lymphocyte recovery was evaluated. Results: The lymphocyte subsets recovered in the following order: NK cells, cytotoxic T cells, B cells, and helper T cells. At 1 month post-transplant, acute graft-versus-host disease was found to contribute significantly to the delay of $CD16^+/56^+$ cell recovery. Younger patients showed delayed recovery of both $CD3^+/CD8^+$ and $CD19^+$ cells. EBV DNAemia had a deleterious impact on the recovery of both $CD3^+$ and $CD3^+/CD4^+$ lymphocytes at 1 year post-transplant. Conclusion: In our pediatric allogeneic HSCT cohort, helper T cells were the last subset to recover. Younger age and EBV DNAemia had a negative impact on the post-transplant recovery of T cells and B cells.

• Korean Journal of Food Science and Technology
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• v.32 no.4
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• pp.881-888
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• 2000

Lactic acid bacteria(LAB) fermented food was prepared with milk or egg white powder(EWP) and added with freeze drying protectant(FDP). 0.2% of Tween 80 or 1% of ascorbate was added to milk sample and 3% of raffinose or 1% of ascorbate was added to EWP sample. Effects of FDP on sensory property, volatile aroma compounds and physical property of LAB fermented food were investigated. In case of non-freeze dried samples, sensory properties of milk sample with ascorbate were slightly better than those of reference sample(milk), while sensory properties of EWP sample or EWP sample with FDP were slightly inferior to reference sample. Sensory properties of all of the freeze dried/reconstituted samples were not different. Sensory properties of milk sample with ascorbate were reduced by freeze drying/reconstitution, while those of sample with ascorbate were not changed. Although all of the volatile aroma compounds were reduced by freeze drying, the residual ratio was slightly different between milk samples and EWP samples. Difference in volatile aroma compounds between milk samples and EWP samples before freeze drying was relatively large, while difference between two sample groups after freeze drying/reconstitution was relatively small. Rheological properties of milk samples were markedly changed by freeze drying/reconstitution, while those of EWP samples were changed slightly.

• Food Engineering Progress
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• v.14 no.1
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• pp.54-59
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• 2010

Instantized thin gruels have been popular to supplement patients who want nutritional, ready-to-eat, and easy-to-use products. In this study, rice-based thin gruels were developed by use of gelatinized rice powder which was manufactured by extrudating rice in a twin-screw extruder. Subsequently, the rice paste from the extruder were dried and ground into fine powder. The gelatinized rice powder was mixed with the powders of various grains, soy beans, nuts, oil seeds, and vegetables to formulate the instantized thin rice gruel with well-balanced nutrients (mixed powder). The mixed powder was granulated to improve reconstitution capability in a fluid bed spray granulator (granulated powder). Lipid and protein contents were higher by 0.9 and 1.9%, respectively, in the granulated powder whereas carbohydrate content was higher by 3.2% in the mixed powder. The calculated dispersibility was 93.7 and 77.0% for the granulated and the mixed powders, respectively. The reconstitution time was 122.3 and 305.3 for the granulated and the mixed powders, respectively. In conclusion, the granulation of the mixed powder improved the dispersibility. This study will be helpful to develop a variety of processed rice products and promote rice process industry.