• BMB Reports
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• v.37 no.4
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• pp.466-473
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• 2004

A new acid deoxyribonuclease (DNase) was purified from the cultured mycelia of Cordyceps sinensis, and designated CSDNase. CSDNase was purified by $(NH_4)_2SO_4$ precipitation, Sephacryl S-100 HR gel filtration, weak anion-exchange HPLC, and gel filtration HPLC. The protein was single-chained, with an apparent molecular mass of ca. 34 kDa, as revealed by SDS-PAGE, and an isoelectric point of 7.05, as estimated by isoelectric focusing. CSDNase acted on both double-stranded (ds) and single- stranded (ss) DNA, but preferentially on dsDNA. The optimum pH of CSDNase was pH 5.5 and its optimum temperature 55. The activity of CSDNase was not dependent on divalent cations, but its enzymic activity was inhibited by high concentration of the cation: $MgCl_2$ above 150 mM, $MnCl_2$ above 200 mM, $ZnCl_2$ above 150 mM, $CaCl_2$ above 200 mM, NaCl above 300 mM, and KCl above 300 mM. CSDNase was found to hydrolyze DNA, and to generate 3-phosphate and 5-OH termini. These results indicate that the nucleolytic properties of CSDNase are essentially the same as those of other well-characterized acid DNases, and that CSDNase is a member of the acid DNase family. To our knowledge, this is the first report of an acid DNase in a fungus.

• Applied Biological Chemistry
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• v.22 no.1
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• pp.1-9
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• 1979

The physical and chemical properties of ${\gamma}-conglycinin$ of soybean (Glycine max) were investigated. The soybean protein extracted from soybean meal using 0.2M NaCl solution at pH 4.5 was passed through a Sephadex G-150 column to isolate 7S globulin. ${\gamma}$-Conglycinin was isolated and purified from the 7S globulin with a DEAE Sephadex A-50 column chromatography. The protein preparation was pure on immunoelectrophoresis, polyacrylamide gel electrophoresis and gel isoelectric fouling. It had an isoelectric point at pH 5.4 and contained 16.12% nitrogen, 4.18% mannose and 1.21% glucosamine. Amino acid composition, in general, shaved that ${\gamma}-conglycinin$ contained higher contents of lysine, dicarboxylic acids and ammonia nitrogen, and lower contents of sulfur-containing amino acids and tryptophan. The subunits of ${\gamma}-conglycinin$ were distributed in the range of pH 4.6-5.5. The subunits located in the pH region of 4.6-5.0 and 5.0-5.5 were glycopeptides (molecular weight of 38,000) and simple peptide (MW of 32,000), respectively.

• Journal of the Korean Wood Science and Technology
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• v.32 no.4
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• pp.58-65
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• 2004

This study was performed to investigate the protease activity from fruit body of Sarcodon aspratus and its features. The specific protease activity was increased with the increasing purification steps, 2.62 times by desalting, 17 times by CMC column chromatography, 113.8 times by DEAE-Sephadex A-50 column chromatography, and 728.3 times by Sephadex G-75 column chromatography. Proteases were identified as two different enzymes having different isoelectric points at pH 4.35 (its recovery rate 8%) and pH 4.7 (its recovery rate 3.5%). Those proteases were purified by 3,025 folds and 3,257 folds in terms of specific activity. Two proteases having different isoelectric points had similar enzymatic properties. This protease was estimated to be 43,000 daltons of molecular weights by SDS-PAGE. This protease with optimum pH 4 was almost stable in the pH range of 4~7. Optimal temperature of protease activity was 40 to 50℃, and the protease activity was completely inhibited at 70℃ for 30 min.

• Journal of the Korean Applied Science and Technology
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• v.10 no.2
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• pp.81-90
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• 1993

The Surfactants composed of acylated aterocollagen which is produced by the acylation of the side chain amino radicals of aterocollagen with an aliphatic acid having 12 to 18 carbon atoms will be discussed in this study. This condensation is done at moderate reaction temperature (<$25^{\circ}C$) in aqueous alkaline solution. The products of this reaction were identified by UV/VIS spectroscopy and infrared spectroscopy. For these compounds, surface active properties and physical properties including isoelectric point, Krafft point, surface tension, critical micelle concentration(cmc), foaming power, viscosity behaviour, water holding capacity, skin irritation and emulsifying power were measured respectively. The experimental results received that the products have a good emulsifying power, excellent water holding capacity while having low skin irritation. Thus, these derivatives will be expected to be used as an emulsifying agent for O/W type cosmetic emulsion.

• Journal of the Korean Society of Tobacco Science
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• v.18 no.1
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• pp.39-48
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• 1996

A storage protein(SP-2) was confirmed in haemolymph during larval-pupal-adult development of tobacco burdworm(Helicoverpa assulta Guenee), and its biochemical characteristics were investigated. The titer of SP-2 showed a peak at mature larva, decreased gradually through the late pupal stage, and became undetectable at adult period. As the results from electrophoretic mnysis, SP-2 was confirmed to be glycolipoprotein(M.W. 332kDa) relatively stable to heat( $\leq$ 68$^{\circ}C$ ). This storage protein was determined to be a tetramer composed of a single subunit with MW of 83kDa, and the isoelectric point was 5.7. The amino acid composition of the SP-2 was characterized. It has relatively high content of methionine and histidine, whereas the contents of tyrosine and phenylalanine were relatively low.

• The Korea Journal of Herbology
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• v.22 no.4
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• pp.21-28
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• 2007

Objectives : Kyenegum has been popularly used long as the digestive. The purpose of this study was to investigate the purification and characteristics of protease obtained from Kyenegum crude enzyme. Methods : Kyenegum protease was purified by precipitation with ammonium sulfate followed by SP-Spharose ion exchange chromatography. The molecular weight of Kyenegum protease was estimated by SDS-PAGE electrophoresis. Results : Kyenegum protease was 3,087 units/mg protein specific activity, 14.5 purification fold and 9.8 % recovery. The molecular weight of protease was estimated to be 18 kDa. The isoelectric point was pI 8.97 and values of Km and Vmax of its were 48 mg/mL and 2 units/min, respectively. Conclusion : The result suggests that the protease obtained from Kyenegum has excellent stability of temperature, acid and collagen substrate specificity.

• Animal cells and systems
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• v.13 no.3
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• pp.345-350
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• 2009

We have isolated and characterized a new insect chicken type (c-type) lysozyme gene from the common cutworm, Spodoptera litura. The full-length cDNA of Spodoptera lysozyme is cloned by rapid amplification of cDNA ends PCR (RACE-PCR). The isolated cDNA consists of 1039 bp including the coding region for a 142-amino acid residue polypeptide, which included a signal peptide of 21-amino acid residue and a mature protein of 121-amino acid residue. The predicted molecular weight of mature lysozyme and its theoretical isoelectric point from amino acid composition is 13964.8 Da and 9.05, respectively. The deduced amino acid sequence of Spodoptera lysozyme gene shows the highest similarity (96.7%) to Spodoptera exigua lysozyme among other lepidopteran species. Amino acid sequence comparison with other the c-type lysozymes, Spodoptera lysozyme has the completely conserved $Glu^{32}$ and $Asp^{50}$ of the active site and eight Cys residues are completely conserved in the same position as that of other lepidopteran lysozymes.

• The Korean Journal of Food And Nutrition
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• v.9 no.3
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• pp.319-324
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• 1996

The functional properties of protein recovered from red crab (Chitinonecetes opiiie) processing in water (RCP) were examined and compared with those of soybean protein isolate at pH 2~10 in water and NaCl solu5ion. The solubilities of RCP and SPI were miniumu at pH 4, the isoelectric point and increased significantly at lower or higher than pH 4. Solubilities in NaCl solution for both proteins decreased with incr NaCl concentration increase at all pH ranges. Emulsion capacity for both proteins was also minimum at pH 4 and increased as protein concentration increased from 2 to 6%. Emulsion capacity of RCP was higher than these of SPI at pH 6∼10 and all protein concentrations. Emulsion stability showed a similar trend to that of emulsion capacity. RCP had higher oft absorption capacity and lower water absorption capacity than SPI.

• Journal of Microbiology and Biotechnology
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• v.6 no.6
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• pp.402-406
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• 1996

Inulin fructotransferase (depolymerizing) (EC 2.4.1.93) was purified 34-fold from the culture broth of Arthrobacter sp. A-6 by using a combination of ammonium sulfate fractionation, DEAE-Sepharose CL-6B chromatography and Sephacryl S-200 gel filtration. The purified enzyme converts inulin into di-D-fructofuranose dianhydride III(DFA III) and small quantities of fructo-oligosaccharides. The temperature and pH optima of the enzyme were $70^{\circ}C$ and 6.0, respectively. Molecular weight of the enzyme was determined to be 49 kDa by 12$%$ SDS-polyacrylamide gel electrophoresis, and 145 kDa by Sephacryl S-200gel filtration. This indicates that the functional inulin fructotransferase of Arthrobacter sp. A-6 has a homomeric trimer structure. The enzyme had an isoelectric point of pH 4.6. The N-terminal amino acid sequence of the purified enzyme subunit was Ala-Asp-Asn-Pro-Asp-Gly(\ulcorner)-Ser-Asn-Met(or Glu)-Tyr-Asp-Val.

• Food Science and Preservation
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• v.17 no.4
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• pp.435-443
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• 2010

Physicochemical and functional properties of collage from skate skin (Raja Kenojei) are examined depending on pH and NaCl concentration in the medium. The solubility decreased as NaCl concentration increased but, turbidity increased as concentration of collagen increased. Oil-holding capacity and water-holding capacity were similar to other fish skin collagens. Emulsion activity, creaming stability, and viscosity were lowest at where pH levels were isoelectric point regions of collagens. However, the higher pH values at 7.0-9.0 caused increasing foam expansion, foam viscosity, and gel strength. These results indicated that collagen from skate skin could be used as a functional ingredient for food and industrial applications.