• Microbiology and Biotechnology Letters
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• v.21 no.4
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• pp.329-335
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• 1993

Ascorbate oxidase activity in various cucumber tissue extracts was highest in young fruit peeling. Cucumber callus was induced from young fruit peeling and callus cell lines were selected for more than 7 months, which porduced high levels of ascorbate oxidase and had a high growth rate. Induction of callus was optimized with Linsmaier-Skoog(LS) medium at 25$^{\circ}C$ in dark phase. Ascorbate oxidase activity reached a maximum at 5 days after transfer to LS basal liquid-medium ant then declined. The enzyme activity in callus cells was stimulated by addition of 10${\mu}$M $CuSO_4$ in the early logarithmic phase of growth. And also, adding 10${\mu}$M $CuSO_4$ at 3rd day 7th day of culture period, ascorbate oxidase activity in callus cells was maintained to high level. Maximum yield of ascorbate oxidase was found at the 25th day by flask shaking culture, but three-fold of ascorbate oxidase activity was obtained at the 16th day by jar fermentation.

• Journal of Microbiology and Biotechnology
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• v.20 no.11
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• pp.1555-1562
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• 2010

Development of the strain and the fermentation process of Hansenula polymorpha was implemented for the production of ${\gamma}$-linolenic acid ($GLA,\;C18:3{\Delta}^{6,9,12}$), an n-6 polyunsaturated fatty acid (PUFA) that has been reported to possess a number of health benefits. The mutated ${\Delta}^6$-desaturase (S213A) gene of Mucor rouxii was expressed in H. polymorpha under the control of the methanol oxidase (MOX) promoter. Without the utilization of methanol, a high-cell-density culture of the yeast recombinant carrying the ${\Delta}^6$-desaturase gene was then achieved by fed-batch fermentation under glycerol-limited conditions. As a result, high levels of the ${\Delta}^6$-desaturated products, octadecadienoic acid ($C18:2{\Delta}^{6,9}$), GLA, and stearidonic acid ($C18:4{\Delta}^{6,9,12,15}$), were accumulated under the derepression conditions. The GLA production was also optimized by adjusting the specific growth rate. The results show that the specific growth rate affected both the lipid content and the fatty acid composition of the GLA-producing recombinant. Among the various specific growth rates tested, the highest GLA concentration of 697 mg/l was obtained in the culture with a specific growth rate of 0.08 /h. Interestingly, the fatty acid profile of the yeast recombinant bearing the Mucor ${\Delta}^6$-desaturase gene was similar to that of blackcurrant oil, with both containing similar proportions of n-3 and n-6 essential fatty acids.

• Korean Journal of Plant Tissue Culture
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• v.21 no.1
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• pp.35-39
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• 1994

Is obtain cell lines showing high level of rice cold tolerance, direct in vitro selection through cold stress on primary pollen callus derived from anther culture was carried out Genotypic difference in callus formation and plant regeneration was recognized Rates of albino was increased along the duration of cold stress. Reciprocal effects were not noticed in anther culturability There was no variants related to rice leaf discoloration in pollen derived lines from parental varieties, regardless of days of cold stress. The regeneration and recombination of rice leaf discoloration in 146 pollen-derived lines, 70 pollen-derived lines from cold stress at $0^{\circ}C$ for 10 days, and 830 F$_2$ plants presented normal distribution curves with skewness in tolerance and no significant difference among 3 populations. Direct in vitro selection for rice cold tolerance through cold stress on primary pollen callus derived from anther culture, therefore, was revealed ineffective as a in vitro technology.

• The Journal of the Convergence on Culture Technology
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• v.9 no.5
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• pp.661-667
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• 2023

Five commercially available edible sweeteners are used as diet products because they can replace sucrose. In studies on the effects on animals and the human body, stability has been proven by excreting-oriented studies with characteristics of animal cells, and accumulation in small amounts has been ignored. On the other hand, plants can absorb, degrade, and accumulate foreign substances, so the effect of degradability and accumulation potential can be studied using plants. Metabolic effects in plants of commercially available saccharin and acesulfame potassium (Ace K) were tested using germinated barley and bean sprouts. In germinated barley and bean sprouts, saccharin and ace K showed inhibitory effects on plant growth in all organs from low concentrations in leaves, stems and roots. In addition, it can be observed that the symptoms of death appear clearly over time, so it can be seen that they are accumulated in the body of the plant. As the accumulated amount increases, the toxic effect increases and the plant reaches a state where it is unable to metabolize, turning black from the tip of the leaf and reaching a state of death. In order to remove the accumulated artificial sweetener, recovery was attempted by culturing in distilled water, but it acts as a substance that is not degraded and dies without avoiding toxicity. Saccharin and ace K cannot be excreted from the cell. Its toxic effects are thought to be persistent, inhibiting growth and eventually leading to cell death.

• Journal of Plant Biotechnology
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• v.35 no.3
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• pp.235-243
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• 2008

Here we describe a procedure for Chinese cabbage protoplast culture and effects of various treatments. Chinese cabbage protoplasts were isolated from different parts of young seedlings as using an enzyme mixture, of which yield was maximized in seven hours around after digestion. The highest rate of initial cell division followed by micro-callus formation was obtained in the medium with 1.0 mg/L 2,4-D, 0.5 mg/L NAA, and 1.0 mg/L BA when the cotyledon-derived protoplasts were cultured. Initiation of cell division and micro-callus proliferation significantly depended upon Chinese cabbage genotype under a same culture circumstances. The micro-calli developed from cotyledon tissue of Norang-Bom cultivar successfully grew toward callus colonies on the solidified medium with 0.2 mg/L zeatin and 0.1 mM spermidine. The callus colonies generated de novo shoots at the maximum frequency of 4.3% on the medium with 5.0 mg/L BA and 1.0 mg/L NM. Our results might be helpful for further studies to enhance the regeneration efficiency in Chinese cabbage protoplast culture.

• Applied Biological Chemistry
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• v.35 no.6
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• pp.443-448
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• 1992

To investigate the production of monoterpenoids by Mentha pipperita cells in suspension culture, effects of media formulation, plant growth hormones, initial pH of the media, and cold stress on the production of essential oil and menthol were analyzed. Among the media employed, Lin-Staba medium resulted in the best essential oil production. Addition of 100 mg/l of yeast extract to the Lin-Staba medium induced the cells to produce large amount of essential oil and high content of menthol (0.39 g/l and 19.6%, respectively). In the effect of plant growth hormone, auxine were more effective than cytokinins. At initial pH of 4.7, oil production was good but menthol content was low. However at pH 5.7 the trend was reversed. When the culture temperature was lowered from $27^{\circ}$ to $10^{\circ}$ during 6 hour-dark period, growth was not changed much but essential oil production and menthol content was increased and reached to 528 mg/l and 21%, respectively.

• Horticultural Science & Technology
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• v.33 no.2
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• pp.251-258
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• 2015

Eurycoma longifolia is an important rare medicinal plant that contains valuable bioactive compounds. In the present study, cell suspension culture of E. longifolia was established for the production of biomass and phenolic compounds. Various medium parameters, such as concentration of auxin, salt strength of the medium, and sucrose and nitrogen concentrations, were optimized for the production of biomass at the flask-scale level. Full strength Murashige and Skoog (MS) medium supplemented with $3.0mg{\cdot}L^{-1}$ naphthaleneacetic acid (NAA), 3% (w/v) sucrose, 0:60 $NH{_4}^+:NO{_3}^-$ was found suitable for biomass accumulation. Based on the optimized flask-scale parameters, cell suspension cultures were established in balloon-type bubble bioreactors, and bioprocess parameters such as inoculum density and aeration rate were optimized. Inoculum density of $50g{\cdot}L^{-1}$ and increasing aeration rate from 0.05 to 0.3 vvm, with increases every 7 days, were suitable for the accumulation of both biomass and phenolic compounds. With the optimized conditions, $14.70g{\cdot}L^{-1}$ dry biomass, $10.33mg{\cdot}g^{-1}$ DW of phenolics and $3.89mg{\cdot}g^{-1}$ DW of flavonoids could be achieved. Phenolics isolated from the cell biomass showed optimal free radical scavenging activity.

• Journal of Life Science
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• v.28 no.8
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• pp.938-944
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• 2018

This study evaluated tyrosinase, elastase inhibitory, and antioxidant activities of fermented Rhododendron micranthum Turcz. extract using a lactic acid bacterium, Lactobacillus rhamnosus. The optimum conditions for fermentation of R. micranthum Turcz. extract were $37^{\circ}C$ and 3% R. micranthum Turcz. extract for 3 days based on the bacterial cell number, total phenolic compounds, DPPH radical scavenging activity, and tyrosinase and elastase inhibitory activity. After culture for 3 days using 3% R. micranthum Turcz. extract, the cell mass of L. rhamnosus reached $5.7{\times}10^9CFU/ml$. The results indicated that R. micranthum Turcz. extract can be used for industrial lactic acid bacteria culture. After fermentation under optimum conditions, the total content of phenolic compounds of the fermented R. micranthum Turcz. extract was 157 GAE mg/ml, and the $IC_{50s}$ of DPPH radical scavenging activity, ABTS radical scavenging activity, and tyrosinase and elastase inhibitory effects were 78.8, 79.8, 329.1, and $449.5{\mu}g/ml$, respectively. The fermented R. micranthum Turcz. extract exhibited 1.2-, 1.3-, 1.5-, 2.4-, and 5.6-fold higher total content of phenolic compounds, DPPH radical scavenging activity, ABTS radical scavenging activity, and tyrosinase and elastase inhibitory effects than the nonfermented R. micranthum Turcz. extract. These results indicated that fermented R. micranthum Turcz. extract using L. rhamnosus can be used for developing new natural functional ingredients for the health food or cosmetic industry.

• Journal of Applied Biological Chemistry
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• v.63 no.3
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• pp.283-290
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• 2020

Oxidative stress is one of the contributors of neurodegenerative disorders including Alzheimer's disease. According to previous studies, Aster yomena (Kitam.) Honda (AY) possesses variable pharmacological activities including anti-coagulant and anti-obesity effect. In this study, we aimed to determine the neuroprotective effect of ethyl acetate fraction from Aster yomena (Kitam.) Honda (EFAY) against oxidative stress. Therefore, we carried out 3-(4,5-dimethylthiazol-2-yl)-2,3-diphenyl tetrazolium bromide, lactate dehydrogenase (LDH), and 2',7'-dichlorofluorescin diacetate assays in SH-SY5Y neuronal cells treated with hydrogen peroxide (H₂O₂). H₂O₂-treated control cells exhibited reduced viability of cells, and increased LDH release and reactive oxygen species (ROS) production compared to normal cells. However, treatment with EFAY restored the cell viability and inhibited LDH release and ROS production. To investigate the underlying mechanisms by which EFAY attenuated neuronal oxidative damage, we measured protein expressions using Western blot analysis. Consequently, it was observed that EFAY down-regulated cyclooxygenase-2 and interleukin-1β protein expressions in H₂O₂-treated SH-SY5Y cells that mediated inflammatory reaction. In addition, apoptosis-related proteins including B-cell lymphoma-2-associated X protein/B-cell lymphoma-2 ratio, cleaved caspase-9, and cleaved-poly (ADP-ribose) polymerase protein expressions were suppressed when H₂O₂-treated cells were exposed to EFAY. Our results indicate that EFAY ameliorated H₂O₂-induced neuronal damage by regulating inflammation and apoptosis. Altogether, AY could be potential therapeutic agent for neurodegenerative diseases.

• Korean Journal of Breeding Science
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• v.49 no.3
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• pp.170-179
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• 2017

Plant molecular farming has attracted a lot of attention lately in the field of mass production of industrially valuable materials by extending application of the plant as a kind of factory concept. Among them, protein expression system using rice(Oryza sativa L.) callus is a technology capable of mass culture and industrialization because of a high expression rate of a target protein. This study was carried out to develop an Agrobacterium-mediated transformation system to increase the utilization of rice callus. The transformation efficiency was improved by using the hand when seeds were de-husked for callus induction. Furthermore, we were possible induction of callus from 6 years old seed smoothly. Selection of the callus contained the target gene was required a cultivation period of at least 3 weeks, and the most efficient selection period was after 6 weeks of culture including one passage. This selection was confirmed that the gene was stably inserted into the genomic DNA of the plant cell by the southern blot analysis and progeny test. Such an efficient selection system of rice callus that can be cultured in the long term will be contribute to the industrialization of useful recombinant proteins using rice.