• Journal of the Korean Society of Food Science and Nutrition
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• v.28 no.2
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• pp.348-352
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• 1999

We have intended to accelerate the secretion of human lysozyme(HLY) with permeabilizing agents from the cultivated cells of the recombinant Saccharomyces cerevisiae. The five agents CaCl2, Tween 80, ethanol, Triton X 100, and cetyltrimethylammonium bromide(CTAB) were used as permeabilizing agents. Treatments of the yeast cell with CaCl2, Tween 80, and ethanol were effective to increase the secretion from the yeast cells. Especially, treatment of 10% ethanol increased the extracellular HLY activity by 38.6% at 30oC for 48 h in culture broth. But Triton X 100 and CTAB unexpectedly didn't play a role in increase of HLY secretion. Recovery of a foreign protein by permeabilizing agents is easier than by osmotic shock, and is less expensive than enzymatic digestion.

• KSBB Journal
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• v.18 no.6
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• pp.435-439
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• 2003

Plant cell culture can be divide into two classes non-organic culture and organic culture. Non-organic culture such as suspension culture has many researches, however organic culture about recombinant protein production has little researches. Recombinant protein produced through organ culture is quite stable and it can make proteins by itself without any grow regulators. Therefore organ culture is much easier than other methods. In this research, we used transformed tobacco seed. At first we germinated the seed then separated stems and leaves from the grown plant. And raised in liquid medium by in vitro vegetative reproduction. Continuing most suitable conditions, we compared the Quantities of recombinant protein from intra cellular with from extra cellular. And adding some permeabilizing agents (Pluronic F-68, Triton X-100, DMSO, PEG8000), we increased the productivity of the recombinant protein.

• Korean Journal of Food Science and Technology
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• v.31 no.6
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• pp.1529-1535
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• 1999

This study was done for the extraction of carotenoid from Phaffia rhodozyma in combination with PEF and other methods. PEF treatment conditions were $30{\sim}80\;kV/cm,\;100{\sim}1000\;Hz\;and\;100{\sim}1000\;{\mu}s$. In order to increase permeability of yeast cell wall, various methods such as freezing-thawing, mechanical treatment, solvents, permeabilizing agents, and yeast cell wall lytic enzyme were used before PEF treatment. The combination of PEF $(50\;kV/cm,\;300\;Hz,\;1000\;{\mu}s)$ and conventional methods such as solvent and freezing-thawing pre-treatment had no effects on the extraction of carotenoid pigments. The extent of extracted carotenoid by the PEF $treatment(50\;kV/cm,\;300\;Hz,\;1000\;{\mu}s)$ combined with yeast cell wall lytic enzyme and mechanical pre-treatment increased 52% and 69.8% more than the sum of that by each treatment, respectively. Permeabilizing agents, especially Tween 20 and capric acid, enhanced the extraction efficiency of carotenoid pigments from P. rhodozyma cells. These results indicated the feasibility for the continuous extracting carotenoid pigments from P. rhodozyma by PEF combined with other permeabilizing methods.

• Korean Journal of Food Science and Technology
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• v.51 no.2
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• pp.188-191
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• 2019

In this study, the poor electro-transformant yield of large DNA in bifidobacteria was improved by increasing the DNA concentration, which was amplified by enhancing electroporation conditions: treating the cell wall weakening agent and cell membrane permeabilizing molecule as well as changing the electrical parameter. In the enhanced conditions, the electroporation frequency increased from 15 to 71 times compared to the initial conditions at the same DNA concentration. As the DNA concentration increased, the difference in the electroporation frequency between the two conditions became greater, and the curve of the enhanced conditions seemed to be saturated with a DNA concentration over $4{\mu}g$. The present study provided a clue to the recovery of the electroporation frequency with large DNA and formulated the relationship between the DNA concentration, the DNA size and the electroporation frequency in bifidobacteria. Therefore, this study will contribute to the expansion of molecular genetic studies of bifidobacteria.

• KSBB Journal
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• v.10 no.3
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• pp.292-297
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• 1995

Studies on the production of Newcastle disease virus(NDV) were carried out to optimize culture conditions such as initial pH, temperature, serum concentration, multiplicity of infection(M.O.I.) as well as the addition of polycation, antioxidant, and DMSO. Initial pH from 7.2 to 8.1 showed little difference on NDV production but the initial pH below 6.8 resulted in the negative effect. The highest NDV titer was obtained at 0.1 M.O.I. In addition, the maximum production of virus was achieved at 2% FBS and optimum temperature was found to be $34^{\circ}C$. Treatment of polycoation increased the virus production. When ascorbic acid was added as an antioxidant, NDV production was also enhanced. Utilization of DMSO, a well-known permeabilizing agent, showed an inhibitory effect on the propagation of NDV.

• KSBB Journal
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• v.31 no.4
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• pp.277-283
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• 2016

In this system, rice cells were genetically modified to express human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig) using RAmy3D promoter induced by sugar depletion. Even though the target protein fused with signal sequence peptide, plant cell wall can be a barrier against secretion of recombinant proteins. Therefore, hCTLA4Ig can be trapped inside cell wall or remained in intracellular space. In this study, to enhance the secretion of hCTLA4Ig from cytoplasm and cell walls into the medium, permeabilizing agents, such as dimethyl sulfoxide (DMSO), Triton X-100 and Tween 20, were applied in transgenic rice cell cultures. When 0.5% (v/v) of DMSO was added in sugar-free medium, intracellullar hCTLA4Ig was increased, on the other hand, the secreted extracellular hCTLA4Ig was lower than that of control. DMSO did not give permeable effects on transgenic rice cell cultures. And Triton X-100 was toxic to rice cells and also did not give enhancing permeability of cells. When 0.05% (v/v) Tween 20 was added in rice cell cultures, however, intracellular hCTLA4Ig was lower than that of control cultures. And the maximum 44.76 mg/L hCTLA4Ig was produced for 10 days after induction, which was 1.4-fold increase compared to that of control cultures. Especially, Tween 20 at 0.05% (v/v) showed the positive effect on the secretion of hCTLA4Ig though the decrease of intracellular hCTLA4Ig. Also, Tween 20 as a non-toxic surfactant did not affect the cell growth, cell viability and protease activity. In conclusion, secretion of hCTLA4Ig could be increased by enhancing permeability of cells regardless of the cell growth, cell viability and protease activity.

• Journal of Oral Medicine and Pain
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• v.32 no.1
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• pp.45-55
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• 2007

Plant extract has attracted considerable interest in oral disease therapy. The present study was performed to observe the antibacterial effect on cariogenic Streptococcus mutans GS5 and Streptococcus sobrinus 6715, and periodontopathic Actinobacillus actinomycetemcomitans Y4 of phytoncide from Chamaecyparis obtusa Sieb. et Zucc employing the measurement of optical density, viable cell counts, and antibiotic sensitivity. The results were as follows: 1. Minimum inhibitory concentration of the phytoncide for S. mutans, S. sobrinus, and A. actinomycetemcomitans was observed to be 0.5%, 1%, and 0.2%, respectively. 2. Minimum bactericidal concentration of the phytoncide for S. mutans, S. sobrinus, and A. actinomycetemcomitans was determined to be 0.5%, 2%, and 0.2%, respectively. 3. The bacteria exposed to the phytoncide become more sensitive to antibiotics. The phytoncide enhanced significantly antibacterial activity of ampicillin against S. mutans and S. sobrinus. It also increased significantly the activity of penicillin and amoxicillin against S. sobrinus. In contrast, the phytoncide augmented the activity of amoxicillin and cefotaxime against A. actinomycetemcomitans but the increase was not statistically significant. The overall results indicate that phytoncide from Chamaecyparis obtusa Sieb. et Zucc used for this study has a strong antibacterial activity against cariogenic and periodontopathic bacteria and that it also has permeabilizing effect on certain antibiotics against these bacteria. Therefore, the phytoncide may be used as a candidate for prevention and therapeutic agent against oral infectious disease including dental caries and periodontal disease.