• Microbiology and Biotechnology Letters
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• v.23 no.2
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• pp.145-149
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• 1995

For the purpose of developing a new biodegradable detergent, we have isolated a gene encoding wide-range temperature applicable alkaline protease from Xanthomonas sp. YL-37 (Lee et al., 1994, Kor. J. Appl. Microbiol. Biotechnol.). An alkaline protease gene was isolated from the gene bank that was prepared from the chromosomal DNA of Xanthomonas sp. YL-37. From the results of agarose gel electrophoresis and a restriction enzyme mapping, a 2.7 kb DNA fragment containing the alkaline protease gene was inserted in the plasmid pUC9. Extracellular activity of a clone having alkaline protease gene was detected on SDS-polyacrylamide gel with activity staining assay. The molecular weight of alkaline protease was determined to be about 64 kDa from 11% SDS-PAGE analysis. Alkaline protease activity, produced from E. coli which harboring the plasmid, showed no difference at reaction temperature 20, 30 and 40$\circ$C, respectively. This result showed that alkaline protease produced from E. coli harboring the plasmid was apparently the same as that of Xanthomonas sp. YL-37.

• Korean Journal of Breeding Science
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• v.40 no.1
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• pp.26-30
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• 2008

Normal soybean seed contains three lipoxygenase isozymes called L-1, L-2, and L-3, respectively, which are responsible for the generation of undesirable grassy-beany flavors. Simple and effective methods for the detection of lipoxygenase isozymes were developed in soybean seeds. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) has been tried in separating these isozymes. It was done effectively on 7.5% separating gel and 4.5% stacking gel. However, no reliable method has been developed specifically for separating L-3, L-13 and L-23. Visual judging methods were based on the bleaching activities of lipoxygenase in contact with methylene blue and ${\beta}$-carotene. Sodium linoleate bleaching method was adopted to determine L-1 and L-2. Carotene bleaching and spectrophotometric methods were used to determine L-3. These systems were very rapid within one minute, furthermore only required a small piece of cotyledon (below 10 mg) and the other part could be used for generation advance after analysis. It was demonstrated that 200 seed samples could be analyzed per day by one laboratory assistant. The combination of visual judging methods and electrophoresis is suitable for breeding programs. It took 6.5 hours for analysis of 100 seed samples by one person.

• Analytical Science and Technology
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• v.8 no.4
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• pp.865-868
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• 1995

The method of matrix-assisted laser desorption ionization mass spectrometry has been used to solve some bioanalytical problems, which is difficult to analyse by general methods. For the selection of proper laser wavelength and matrices, eight matriees was used with laser wavelength of 226 and 355nm. The result shows that with wavelength of 355nm better results could be obtained with most of the matrices. The molecular weight of eytochrome C, which was seperated by gel electrophoresis and electro-blotted onto NC membrane is determined by MALDI. The accuracy is better than 0.1%, which is much higher than that of SDS-PAGE. Protein mixture extracted from crude peanut oil is directly determined by MALDI. The molecuiar weight of its three components are determined, and the result also demonstrated that these proteins are in free manner. As proteins arc in 2S bond, with the traditional method, SDS-PAGE, it is not able to decide whether protein exists in combination mode or in free manner. In the technique of two phase aquesous solution, which is used for separating biomaterials, water soluble polymers stained with dyes are used in this technique. By the use of MALDI the number or the dye molecules react with the polymer PEG molecule are determined, and that is difficult to determined by other methods.

• Archives of Pharmacal Research
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• v.21 no.5
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• pp.521-526
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• 1998

The physicochmical properties of recombinant hepatitis B surface antigen (r-HBsAg), which was expressed in C127 mammalian cell were studied. Using roller bottle culture in DMEM supplemented with fetal bovine serum, 10-15 mg/L of r-HBsAg was produced with about 31% of purification yield. The purity of r-HBsAg by HPLC was 99.8% and electron microscopic examination showed homogeneous spherical particle with 22 nm in diameter, a morphological characteristic of HBsAg. The density of r-HBsAg by CsCI density gradient method was 1.19g/ml and the isoelectric point by Mono $P^{TM}$ HR 5/20 column was 4.6. The analysis of subunit protein pattern using SDS-PAGE followed by scanning densitometry gave 81.3% of S protein and 18.7% of pre-S protein. fluorophore-assisted-carbohydrate-electrophoresis analysis showed the relative amount of carbohydrate to protein was 1.7% and it smajr component was N-acetyl glucosamine, which was about 39% of total carbohydrate. The relative amount of lipid to protein determined by vanillin phosphoric acid method was 32.5% and its major component was phospholipid, which was about 70% of total lipid. The physicochemical properties of C127 mammalian cell-derved r-HBsAg are similar to those of p-HBsAg, suggesting that the r-HBsAg can be used in developing a new preventive vaccine against hepatitis B.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.28 no.6
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• pp.753-760
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• 1995

This study was conducted to determine the serum vitellogenin (VTG) concentration changes during the ovulation period in elkhorn sculpin, Alcichthys alcicornis. The results of sepacryl S-300 showed that the molecular weight of VTG could be 380,000. SDS-polyacrylamide gel electrophoresis (SDS-PAGE) analysis may indicate that the purified VTG consists of three subunits with molecular weights of 180,000, 118,000 and 85,000, respectively. Yolk protein purified from the egg extracts was eluted on an equilibrated sephacryl S-300 column, and its molecular weight was estimated 250,000. The precipitation lines of the female serum against the antiserum of the egg extracts were fused completely by immunoelectrophoresis and immunodiffusion analysis. VTG was detected in the serum, and hepatocytes from males injected with $17\beta-estradiol\;(E_2).$ Furthermore, VTG was immunochemically similar to yolk proteins. The concentration of VTG was high before ovulation $(9.80\pm0.81-11.02\pm0.09 mg/ml),$ and then decreased rapidly after ovulation $(less\;than\;6.19\pm0.59 mg/ml).$ This study suggested that VTG was synthesized in the liver by the action of $E_2$ and released to blood, and then incorporated into oocytes.

• Journal of Animal Science and Technology
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• v.63 no.2
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• pp.405-416
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• 2021

The aim of the study was to determine the effect of whole milk powder (WMP) as heterologous proteins on chicken breast emulsion-type sausages. The quality properties of WMP on such chicken breast emulsion-type sausages were investigated by measuring the proximate composition, pH, color, cooking yield, protein solubility, and by applying other methods, such as texture profile analysis (TPA), microphotograph, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and electronic nose. The crude fat, protein, and ash contents of 15% WMP samples were significantly higher than the control samples (p < 0.05). The redness of the cooked samples significantly increased with an increase in the WMP contents (p < 0.05). The cooking yield of WMP treated samples was significantly higher than the control sample (p < 0.05). Additionally, the hardness, gumminess, and chewiness of WMP treated samples were significantly higher than the control sample (p < 0.05). The sarcoplasmic and myofibrillar proteins of samples containing 15% WMP were significantly higher than the control samples (p < 0.05). The result of SDS-PAGE showed that the C protein, sarcoplasmic protein, actin, and tropomyosin increased with an increase in the WMP contents. The principal component analysis plot of WMP-treated samples was clearly different from that of the control samples. Based on these results, it was predicted that WMP could be useful as heterologous protein on emulsion-type sausage.

• Korean Journal of Agricultural Science
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• v.45 no.4
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• pp.635-643
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• 2018

Bovine milk is widely consumed by humans and is a primary ingredient of dairy foods. Proteomic approaches have the potential to elucidate complex milk proteins and have been used to study milk of various species. Here, we performed a proteomic analysis using 2-dimensional electrophoresis (2-DE) and matrix assisted laser desorption ionization-time of flight mass spectrometer (MALDI-TOF MS) to identify whey proteins in bovine milk obtained soon after parturition (bovine early milk). The major casein proteins were removed, and the whey proteins were analyzed with 2-dimensional polyacrylamide gel electrophoresis (2-D PAGE). The whey proteins (2 mg) were separated by pI and molecular weight across pH ranges of 3.0 - 10.0 and 4.0 - 7.0. The 2-DE gels held about 300 to 700 detectable protein spots. We randomly picked 12 and nine spots that were consistently expressed in the pH 3.0 - 10.0 and pH 4.0 - 7.0 ranges, respectively. Following MALDI-TOF MS analysis, the 21 randomly selected proteins included proteins known to be present in bovine milk, such as albumin, lactoferrin, serum albumin precursor, T cell receptor, polymeric immunoglobulin receptor, pancreatic trypsin inhibitor, aldehyde oxidase and microglobulin. These proteins have major functions in immune responses, metabolism and protein binding. In summary, we herein identified both known and novel whey proteins present in bovine early milk, and our sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis revealed their expression pattern.

• Journal of Ginseng Research
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• v.23 no.1 s.53
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• pp.55-59
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• 1999

For the first time we have developed a simple method for conjugation between ginsenoside Rf (Rf) and bovine serum albumin (BSA) or ovalbumin (OVA) by a periodate oxidation method. The Rf-BSA conjugate is confirmed by ultraviolate (UV) scanning, thin layer chromatography (TLC), and SDS-gel electrophoresis. In UV scanning Rf showed three small peaks approximately at 230, 265 and 280 nm. BSA showed a peak at 280 nm. The Rf-BSA conjugate showed right shifted three peaks that BSA alone did not show. In TLC analysis, the Rf-BSA conjugate did not show mobility in silica gel but showed a slight stream of trace. In SDS-gel electrophoresis, Rf-BSA conjugate show a slight less mobility than BSA alone. Rf-OVA conjugate also showed similar patterns with Rf-BSA conjugate. These results demonstrated that periodate oxidation method could be used to produce a stable Rf-BSA or Rf-OV A conjugate and also could be applied for other ginsenoside(s).

• The Korean Journal of Pesticide Science
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• v.2 no.1
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• pp.91-96
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• 1998

For the expression of the cry11Aa gene highly toxic to dipteran insects, we constructed two cyanobacteria-Escherichia coli(E. coli) shuttle vectors, pCYASK5-l and pCYASK5-2. These vectors were transformed into E. coli and selected with kanamycin. The expression of the cry11Aa gene in E. coli was characterized by SDS-polyacrylamide gel electrophoresis and Western blot analysis. Two E. coli transformants harboring pCYASK5-1 and pCYASK5-2 expressed the cry11Aa gene in size of 72 kDa and 64 kDa, respectively and showed over 89% mortality against Culex pipiens larvae.

• Journal of Animal Science and Technology
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• v.63 no.2
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• pp.417-425
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• 2021

The use of edible insects to replace meat protein is important to ensure future global food security. However, processed foods using edible insects require development to enhance consumer perception. Here, we examined the physicochemical characteristics and rheological properties of emulsions prepared from different edible insect larvae. Three edible insect species (Tenebrio molitor, Allomyrina dichotoma and Protaetia brevitarsis seulensis) were used to prepare larval emulsions that were formulated with 65% of insect larvae, 20% of pork back fat, and 15% ice. The A. dichotoma emulsion had the highest pH and lightness, redness, and yellowness values, while the T. molitor emulsion had the lowest pH and lightness, redness, and yellowness values. The T. molitor emulsion had the highest hardness, gumminess, chewiness, and apparent viscosity values but the lowest springiness and cohesiveness values. According to the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis, T. molitor had the thickest bands, followed by P. brevitarsis seulensis. The differential scanning calorimetry distributions for the T. molitor and A. dichotoma emulsions showed one peak, while that of the P. brevitarsis seulensis emulsion had two peaks. The collective results suggest that T. molitor was the most suitable candidate (of the three tested species) for use as a meat replacement in terms of its physicochemical and rheological properties. It is important that such properties of insect-based emulsions are maintained using various technologies.