• BMB Reports
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• v.48 no.8
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• pp.438-444
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• 2015

Lysosomal storage diseases (LSDs) are a group of inherent diseases characterized by massive accumulation of undigested compounds in lysosomes, which is caused by genetic defects resulting in the deficiency of a lysosomal hydrolase. Currently, enzyme replacement therapy has been successfully used for treatment of 7 LSDs with 10 approved therapeutic enzymes whereas new approaches such as pharmacological chaperones and gene therapy still await evaluation in clinical trials. While therapeutic enzymes for Gaucher disease have N-glycans with terminal mannose residues for targeting to macrophages, the others require N-glycans containing mannose-6-phosphates that are recognized by mannose-6-phosphate receptors on the plasma membrane for cellular uptake and targeting to lysosomes. Due to the fact that efficient lysosomal delivery of therapeutic enzymes is essential for the clearance of accumulated compounds, the suitable glycan structure and its high content are key factors for efficient therapeutic efficacy. Therefore, glycan remodeling strategies to improve lysosomal targeting and tissue distribution have been highlighted. This review describes the glycan structures that are important for lysosomal targeting and provides information on recent glyco-engineering technologies for the development of therapeutic enzymes with improved efficacy. [BMB Reports 2015; 48(8): 438-444]

• Molecules and Cells
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• v.25 no.4
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• pp.494-503
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• 2008

Many therapeutic glycoproteins have been successfully generated in plants. Plants have advantages regarding practical and economic concerns, and safety of protein production over other existing systems. However, plants are not ideal expression systems for the production of biopharmaceutical proteins, due to the fact that they are incapable of the authentic human N-glycosylation process. The majority of therapeutic proteins are glycoproteins which harbor N-glycans, which are often essential for their stability, folding, and biological activity. Thus, several glyco-engineering strategies have emerged for the tailor-making of N-glycosylation in plants, including glycoprotein subcellular targeting, the inhibition of plant specific glycosyltranferases, or the addition of human specific glycosyltransferases. This article focuses on plant N-glycosylation structure, glycosylation variation in plant cell, plant expression system of glycoproteins, and impact of glycosylation on immunological function. Furthermore, plant glyco-engineering techniques currently being developed to overcome the limitations of plant expression systems in the production of therapeutic glycoproteins will be discussed in this review.

• Korean Journal of Materials Research
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• v.9 no.5
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• pp.467-472
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• 1999

The isothermal cure behavior of diglycidyl ether of bisphenol A(DGEBA) 4,4'-methylene dianiline(MDA) system with various contents of neopentyl glycol(NPG) has been analyzed by differential scanning calorimetry(DSC). TO increase the cure rate of DGEBA/MDA system, NPG was introduced as an accelerator. Regardless of the NPG content, the shape of the conversion curves showed sigmoid indication that DGEBA/MDA/NPG system followed autocatalytic cure reaction. The cure reaction of DGEBA/MDA system increased with the increment of NPG content and it was due to the catalytic role of hydroxyl groups of NPG.

• Microbiology and Biotechnology Letters
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• v.21 no.4
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• pp.336-341
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• 1993

In metal-affinify aqueous two-phase systems, protein partitioning is affected by a variety of parameters such as pH, the number of surface-accessible histidines, and the amount and partition coefficient of metallated polythylene glyco(PEG) ligand. To enhance partitioning of proteins with surface-accessible histidines, we have synthesized and used a (Cu(II)-ininodiacetic acid)$_2$-PEG20,000($Cu(II)_2IDA_2$-PEG20,000) as well as Cu(II)IDA-PEG5,000 as an affinity ligand. The partition coefficient of $Cu(II)_2-IDA_2$-PEG20,000 in a PEG5,000/dextran two-phase system was 30.1, which corresponded to a 3.8-fold increase over that of Cu(II)IDA-PEG5,000. The partitioning experiments were performed on four proteins, horse cytochrome c, S. cerevisiae cytochrome c, horse myoglobin, and sheep myoglobin. Partitioning of proteins which convey surface-accessible histidines was enhanced dramatically by the addition of $Cu(II)_2IDA_2$-PEG20,000 ligand. These results demonstrate that enhanced partitioning of metal-binding proteins in an aqueous two -phase system can by achieved by using an appropriate metallated PEG ligand.

• Korean Journal of Food Science and Technology
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• v.21 no.4
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• pp.505-510
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• 1989

Red muscle and white muscle were separated from bovine, porcine and poultry skeletal muscles, respectively. Intramuscular lipids were extracted and fractionated to neutral-, glyco- and phospho-lipid by silica gel chromagraphy and then fatty acid composition were analyzed with gas chromatography. The results obtained were as follows; Total lipid content of red muscle was higher than that of white muscle in case of beef and chicken. In pork, however, total lipid content of white muscle was higher than red muscle The content ratio of neutral lipid to phospholipid revealed a number of distinctions between red and white muscle among animals. There were noteworthy differences in respect of polyunsaturated fatty acid. The intramuscular fat of pork had the higher content of highly polyunsaturated fatty acid such as arachidonic acid in contrast to beef.

• Journal of Plant Biotechnology
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• v.49 no.1
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• pp.39-45
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• 2022

Conserved domains are defined as recurring units in molecular evolution and are commonly used to interpret the molecular function and biochemical structure of proteins. Herein, the ADP-glucose pyrophosphorylase (AGPase) amino acid sequences of three species of the Ipomoea genus [Ipomoea trifida, I. triloba, and I. batatas (L.) Lam. (sweetpotato)] were identified to investigate their physicochemical and biochemical characteristics. The molecular weight, isoelectric point, instability index, and grand average of hyropathy markedly differed among the three species. The aliphatic index values of sweetpotato AGPase proteins were higher in the small subunit than in the large subunit. The AGPase proteins from sweetpotato were found to contain an LbH_G1P_AT_C domain in the C-terminal region and various domains (NTP_transferase, ADP_Glucose_PP, or Glyco_tranf_GTA) in the N-terminal region. Conversely, most of its two relatives (I. trifida and I. triloba) were found to only contain the NTP_transferase domain in the N-terminal region. These findings suggested that these conserved domains were species-specific and related to the subunit types of AGPase proteins. The study may enable research on the AGPase-related specific characteristics of sweetpotatoes that do not exist in the other two species, such as starch metabolism and tuberization mechanism.

• Membrane Journal
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• v.24 no.2
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• pp.107-112
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• 2014

The composite membranes were prepared on the surface of hydrophobic porous poly (vinylidene fluoride) (PVDF) hollow fiber membranes through the interfacial polymerization. The preparation variables were the concentrations of piperazine (PIP), trimesoyl chloride (TMC) and the contents of polyethylene glyco l (PEG). The separation characterization of the resulting membranes were carried out for aqueous 100 ppm solution of NaCl, $CaSO_4$, and $MgCl_2$ and also mixed 300 ppm solution of NaCl and $CaSO_4$ in terms of the flux and rejection. Both the flux and rejection were the highest when the interfacial polymerization was conducted using TMC. When TMC concentration was 0.1 wt%, the flux and rejection were shown 48.3 LMH ($L/m^2{\cdot}hr$) and 59%, respectively. To improve the flux, the annealing post-treatment and the addition of PEG into piperazine were done. As expected, the overall flux was enhanced while the rejection was reduced.

• KSBB Journal
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• v.21 no.4
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• pp.248-254
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• 2006

Animal cell culture industry has a large market and an exponential growth rate among biological industry field. Chines hamster ovary(CHO) cells are the most widely used cell lines for recombinant protein production. They can avoid infection from polio, herpes, hepatitis B, HIV, measles, adenovirus and etc. Moreover it is easy to transfection recombinant genes and possible to suspension culture. Serum free media is one of the most important factor of protein production. Because serum has problems. Serum is not defined the contents until now, it has a number of proteins, lipids, carbohydrates and unknown molecules that cause of risk involve in infection and high cost of product purification. CHO cell line cultured using serum free media were the basis of a very successful method to produce(glyco-)protein in mammalian cells, which are then used as pharmaceutical products. Also, the low protein content of the developed medium facilitates downstream processing and product purification. But non-adapted CHO cells have a limit of proliferation cultured using serum free media and it takes very long time to adapt non-adapted cells to serum free media. There are a number of causes of a limit of proliferation using serum free media. Absence of growth factors and growth stimulating molecules is a major factor of the reasons. It makes growth signals and moves cell cycle. And increase of cellular stress is another reason. It induces increase of intraceullar ROS concentration. The purpose of this study is about improvement of proliferation capacity of non-adapted CHO cells cultured using serum free media without adaptation process.