• Title/Summary/Keyword: Optimal culture

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Establishment of Acclimatization System and Growth Characteristics for Regenerated Plants of Oplopanax elatus Nakai (땃두릅나무 재분화 유식물체의 순화 체계 및 생육 특성)

  • Seong, Eun Soo;Yoo, Ji Hye;Kim, Hee Young;Choi, Hye Lim;Seo, Ji Won;Hwang, Myeong Ha;Kim, Myong Jo;Yu, Chang Yeon
    • Korean Journal of Medicinal Crop Science
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    • v.27 no.6
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    • pp.397-403
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    • 2019
  • Background: Oplopanax elatus is widely distributed at high altitudes (about 1,100 m) in China, Russia and Korea. It is hard to propagate, breed, and difficult to grow. Hence, it has been designated as a rare and endangered medicinal plant. A study was conducted to establish a system for large scale seedling production of Oplopanax elatus in vitro and to find the ideal environment for its seedling growth. Methods and Results: In this study, the explants produced under in vitro conditions during our previous study were grouped into three categories (under 10 mm, 10 mm - 30 mm and above 30 mm) based on plant height and were transferred to the growth-chamber and greenhouse for two weeks in each setting for acclimatization. The plantlet category of above 30 mm showed good performance, and was further evaluated under three acclimatization methods as follows: three different growth media (commercial soil, commercial soil + perlite, commercial soil + sand), four shading levels (0%, 50%, 70%, 90%) and four altitude levels (157 m, 218 m, 601 m, 870 m) in Gangwon province of South Korea. As results, O. elatus seedlings showed better growth characteristics at 870 m of altitude, 70% shading level and in the commercial soil compared to other treatments. Conclusions: The regenerated seedlings of Oplopanax elatus obtained through plant tissue culture would be advantageous for use in large scale seedling production systems paired with a good acclimation method. For obtaining optimal results, it is recommended that seedling be acclimatized in a high altitude environment.

Effect of kinds of medium and plant growth regulators for plantlets regeneration by bud culture in Disporum sessile (윤판나물(Disporum sessile) 아배양에 의한 식물체 재분화에 영향하는 배지 종류 및 생장조절물질 효과)

  • Lee, Na-Nyum;Kim, Ji-Ah;Kim, Tae Dong;Kim, Yong Wook
    • Journal of Plant Biotechnology
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    • v.44 no.1
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    • pp.42-48
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    • 2017
  • This study was conducted to establish the optimal condition for in vitro propagation of Disporum sessile. In the experiments with different kinds of media (MS, B5 and WPM) and explant types for shoot induction and elongation, the highest number of shoot inductions (2.5/explant) was shown when the axillary bud explants were cultured on MS medium without plant growth regulators (PGRs). The best shoot elongation (7.2 cm) was also obtained when the apical buds were cultured on MS medium. The effect of BA pulse treatments with in vitro shoots was also examined. The highest in shoot induction (2.29/explant) and elongation (7.28 cm) was observed when the shoots from axillary buds were cultured on the media without PGRs. However, it decreased with increasing duration of BA pulse-treatment. The highest rooting rate (100%) and number of root inductions (21.3/explant) were achieved with 1.0 mg/L indole-3-butyric acid (IBA) treatment, whereas no differences were observed by different shoot types. The regenerated in vitro plantlets were hardened and successfully established in soil.

Optimal Fermentation Conditions of Hyaluronidase Inhibition Activity on Asparagus cochinchinensis Merrill by Weissella cibaria

  • Kim, Minji;Kim, Won-Baek;Koo, Kyoung Yoon;Kim, Bo Ram;Kim, Doohyun;Lee, Seoyoun;Son, Hong Joo;Hwang, Dae Youn;Kim, Dong Seob;Lee, Chung Yeoul;Lee, Heeseob
    • Journal of Microbiology and Biotechnology
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    • v.27 no.4
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    • pp.701-708
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    • 2017
  • This study was conducted to evaluate the hyaluronidase (HAase) inhibition activity of Asparagus cochinchinesis (AC) extracts following fermentation by Weissella cibaria through response surface methodology. To optimize the HAase inhibition activity, a central composite design was introduced based on four variables: the concentration of AC extract ($X_1$: 1-5%), amount of starter culture ($X_2$: 1-5%), pH ($X_3$: 4-8), and fermentation time ($X_4$: 0-10 days). The experimental data were fitted to quadratic regression equations, the accuracy of the equations was analyzed by ANOVA, and the regression coefficients for the surface quadratic model of HAase inhibition activity in the fermented AC extract were estimated by the F test and the corresponding p values. The HAase inhibition activity indicated that fermentation time was most significant among the parameters within the conditions tested. To validate the model, two different conditions among those generated by the Design Expert program were selected. Under both conditions, predicted and experimental data agreed well. Moreover, the content of protodioscin (a well-known compound related to anti-inflammation activity) was elevated after fermentation of the AC extract at the optimized fermentation condition.

Optimizing the Production of 5-Aminolevulinic Acid by Recombinant Escherichia coli Containing the Rhodobacter capsulatus hemA Gene (Rhodobacter capsulatus hemA 유전자 발현 대장균에 의한 5-Aminolevulinic Acid 생산의 최적화)

  • Yang, Dong-Soo;Park, Moon-Won;Lim, Soo-Jin;Kim, Min-Jeong;Shin, Yu-Ri;Park, Chan-Soo;Hyun, Young;Kang, Dae-Kyung
    • Microbiology and Biotechnology Letters
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    • v.37 no.2
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    • pp.153-159
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    • 2009
  • Recombinant Escherichia coli BLR(DE3) harboring the hemA gene from Rhodobacter capsulatus under the control of a constitutive promoter, which we constructed previously, was used for the extracellular production of 5-aminolevulinic acid (ALA). The effects of several factors on ALA production were investigated in flask culture. ALA production by the recombinant E. coli was more efficient at $30^{\circ}C$ than $37^{\circ}C$. The glycine concentration had an important effect on cell growth. Glycine and succinic acid concentration of 5-10 and 10-20 g/L, respectively, resulted in high ALA production. In addition, the partial replacement of succinic acid by sodium glutamate increased the ALA production. The ALA production was inhibited by the presence of glucose in the medium. Using the optimal conditions, an ALA concentration of 8.2 g/L was achieved in jar fermentation without an added inducer or ALA dehydratase inhibitor; this is the highest reported concentration.

Characteristics of the protease from the extreme halophile, Halobacterium sp. (고도 호염성 Halobacterium sp.가 생산하는 protease의 특성)

  • Ahan, Young-Seok;Kim, Chan-Jo;Choi, Seong-Hyun
    • Applied Biological Chemistry
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    • v.33 no.4
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    • pp.337-342
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    • 1990
  • The pretense from Halobacterium sp. was purified by ethanol precipitation and gel filtration on Sephadex G-75 and G-100. The purified enzyme was found to be homogeneous by polyacrylamide gel electrophoresis It's specific activity was 364units/mg protein and yield was 14% of the total activity of the culture filtrate. The Km value against casein was determined to be $4.2{\times}10^{-4}M$ by Lineweaver-Burk plot The optimal temperature and pH for the enzyme activity were $35^{\circ}C$ and pH 8.0, respectively. The enzyme was stable from 5.0 to 11.0 at relatively wide range of pH but was inactivated at the temperature above $50^{\circ}C$. $Ca^{2+}$ and $Mg^{2+}$ appeared to react as activators whereas $Fe^{3+},\;Zn^{2+},\;Cu^{2+},\;Hg^{2+}\;and\;Cd^{2+}$ as inhibitors. The enzyme activity reduced with increasing the concentration of NaCl : the apparent activity with 2M NaCl was 65% as compared with that without the salt However the enzyme was unstable without salts : the activity was lost when dialyzed against distilled water for 2hr, whereas maintained against 0.1M solution of $CaCl_2$ for 6hr.

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Optimization of Production Conditions of Biosurfactant from Bacillus sp. and its Purification (Bacillus sp.에 의한 생물계면활성제의 생산 및 그의 성질)

  • Kim, Jin-Sook;Song, Hee-Sang;Chung, Nam-Hyun;Bang, Won-Gi
    • Applied Biological Chemistry
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    • v.48 no.2
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    • pp.109-114
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    • 2005
  • A bacterium capable of emulsifying hydrocarbon, n-hexadecane, and decreasing surface tension of the culture media using oil collapsing method was isolated. The bacterium was partially identified as Bacillus sp. and named BJS-51. n-Hexadecane was the most effective carbon source for production of biosurfactant. Surface tension was decreased from 76 dyne/cm to 31 dyne/cm and CMD (critical micelle dilution) had the highest value of 5.7 at 3% n-hexadecane. Ammonium phosphate was the most effective nitrogen source, when C/N ratio was 60, surface tension and CMD were 29 dyne/cm and 9.2, respectively. Optimum pH and temperature were 7.2 and $30^{\circ}C$, respectively. Produced biosurfactant was extracted and purified using organic solvent extraction method and preparative HPLC systems. After analysis by various color reaction, this biosurfactant was identified as lipopolysaccharide. Surface tension and CMC (critical micelle concentration) of purified biosurfactant were 27 dyne/cm and 0.08 g/l, repectively. CMD was 9.2, so the yield of biosurfactant was about 0.74 g/l at the optimal conditions. The biosurfactant was very stable at wide range of $pH\;2{\sim}12$ with surface tension $29{\sim}31\;dyne/cm$ and showed $29{\sim}30\;dyne/cm$ of surface tension after heat treatment at $100^{\circ}C$ for 60 min.

EFFECTS OF FLUORIDE MOUTHRINSE ON CELL ACTIVITY OF GINGIVAL FIBROBLASTS OF CHILDREN (불소양치용액이 소아 치은 섬유아세포의 세포활성에 미치는 영향에 관한 연구)

  • Lee, Dong-Hyun;Lee, Kwang-Hee
    • Journal of the korean academy of Pediatric Dentistry
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    • v.24 no.1
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    • pp.204-219
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    • 1997
  • The use of fluoride is one of the most effective methods for caries prevention. Fluoridation of public water supply has been recognized, for many years, as an effective way to reduce dental caries. The fluoride supplement has been recommended when the natural fluoride was unavailable or below the optimal range. However the mechanism of caries prevention by fluoride has not yet been clarified and it is well known that an overdose of fluoride results inacute and chronic toxicity, especially dental fluorosis. Fluoride mouthrinsing solution is widely used in dentistry due to its effectiveness in carrying anticariogenic action. Understanding the effects of fluoride mouthrinsing solution on human gingival fibroblasts will provide the safety rationale for its use during the caries preventive therapy. The purpose of this study was to evaluate the cytotoxic effect of fluoride mouthrinsing solution on the human gingival fibroblast in vitro. The human gingival fibroblasts were cultured from healthy gingiva on the extracted deciduous teeth of children. Cells were inoculated into a 24-well plate with $1{\times}10^4cells/well$ of medium at $37^{\circ}C$, 100% humidity, 5% $CO_2$ incubator for 24 hours. And the cells were counted by using the hemocytometer at each designed study. Human gingival fibroblasts were cultured in growth medium after one minute application range of 0.02%-0.2% NaF solution and 0.1% $SnF_2$ solution. The cells used in this study were between fifth to eighth passage number. The cell morphology was examined by inverted microscope and cell proliferation was measured by incorporating $[^3H]$-thymidine into DNA. DNA synthesis by human gingival fibroblasts was assessed by $[^3H]$-thymidine uptake assays while the cell activity was measured by MTT assay. Each concentrated fluoride mouthrinsing solution was estimated for its biocompatability with fibroblasts by the tissue culture technique. The results of this study were as follows : 1. It was observed that at 0.05%, 0.2% NaF mouthrinsing solution the cytoplasmic processes became globular. When 0.1% $SnF_2$ mouthrinsing solution was applied, the cytoplasmic process and cell morphology were disappeared. 2. DNA synthetic activity was reduced regardless of the concentration of the fluoride mouthrinsing solution. However, the result is statistically insignificant except 0.1% $SnF_2$ mouthrinsing solution(p<0.05). 3. Our results indicate that 0.02%, 0.05% concentrations of NaF mouthrinsing solution caused minimal cytotoxicity. But 0.2% NaF and 0.1% $SnF_2$ concentration were a significant difference between the cell activity in the experimental group and control group (p<0.05). 4. After appling 0.05% & 0.02% NaF fluoride mouthrinsing solution, cell activity was restored to the control groups level according to incubating time. The results suggest that direct exposure to fluoride solution inhibits gingival fibroblast activity. Therefore, for the most effective use of fluoride use, lowering the concentration of fluoride mouthrinsing is advisable because it maintains biocompatability and free ion in the oral fluid.

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Screening of Non-Biogenic-Amine-Producing Bacillus subtilis and Medium Optimization for Improving Biomass by the Response Surface Methodology (바이오제닉 아민 비생성 Bacillus subtilis의 선별 및 반응표면 분석법에 의한 균체량 증가를 위한 배지 최적화)

  • Yang, Hee-Jong;Jeong, Su-Ji;Jeong, Seong-Yeop;Heo, Ju-Hee;Choi, Nack-Shick;Jeong, Do-Youn
    • Journal of Life Science
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    • v.26 no.5
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    • pp.571-583
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    • 2016
  • Biogenic amines are produced primarily by microorganisms found in fermented foods and are often implicated in poisoning incidents in humans. In this study, 620 strains of microorganisms were isolated from traditional Korean fermented food in Sunchang in order to screen for non-biogenicamine-producing microorganisms present in these foods. One strain was identified and named Bacillus subtilis SCJ1, by using 16S rRNA sequencing and biochemical characterization. We investigated the cell growth of this organism in order to understand its potential for industrial application. To this end, we optimized the culture medium constituents by using the response surface methodology. The Plackett-Burman experimental design was used for screening of the medium constituents, such as molasses, yeast extract and peptone, for improving cell growth. In order to determine the optimal concentration of each constituent, we used a central composite design. Consequently, the optimized concentrations of molasses, yeast extract and peptone were predicted to be 27.5 g/l, 7.5 g/l and 17.5 g/l, respectively. By model verification, we confirmed that a 1.49-fold increase in dry cell weight compared to the basal medium-from 1.32 g/l, to 1.9722 g/l-was achieved.

Effects of Natural Compounds from Various Plant Eradicate the Persister Cell of Edwardsiella tarda Treated with Antibiotics of Florfenicol and Amoxicillin (천연 식물 추출물 첨가에 의한 어류 에드워드증(Edwardsiellosis) 발생균인 Edwardsiella tarda에 항생제 투여로 생성되는 persister cell 저감 효과)

  • Kim, Na-Kyoung;Kweon, Dae-Hyuk;Kim, Sung-Koo
    • Journal of Life Science
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    • v.22 no.6
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    • pp.788-793
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    • 2012
  • High concentration of antibiotics has been used to treat the outbreak of edwardsiellosis caused by Edwardsiella tarda in aquaculture. However, not all of the bacteria have been killed with high concentrations of antibiotics treatment by the formation of persister cells with a dormant state. The main objective of this study was to kill persister cell using antibiotics with the addition of natural plant compounds. Antibiotics used in this study consist of 100 mg/ml florfenicol and 100 mg/ml amoxicillin. Ten natural plant compounds with persister cell inhibitor activity to E. coli were obtained from Protein Engineering and Systems Biology Lab. of Sungkyunkwan University. The persister cell inhibition activities of those natural plant compounds were evaluated in test tube. Concentrations of the antibiotics were in the ranges of 25~200 ${\mu}g/ml$. The persister cell formation was observed after 16 hours of culture. Persister cells were killed by antibiotics with natural plant compounds. Among ten natural plant compounds, Gynostemma pentaphyllum, Mallotus japonicus, and Orixa japonica showed persister cell formation inhibition activities. The optimal concentrations of G. pentaphyllum, M. japonicus, and O. japonica for the inhibitor of persister cell formation were 100 ${\mu}g/ml$, 100 ${\mu}g/ml$, and 200 ${\mu}g/ml$, respectively. In vivo study was carried out to evaluate the effect of the antibiotics with natural plant compounds using aquacultural fish, olive flounder, as test animals. G. pentaphyllum, M. japonicus, and O. japonica of 30 ${\mu}g/ml$, 10 ${\mu}g/ml$, and 10 ${\mu}g/ml$ with antibiotics reduced cumulative mortalities, showing the effectiveness of persister cell inhibition.

Effects of Fertilization Methods on the Growth and Physiological Characteristics of $Larix$ $kaempferi$ Seedlings in the Container Nursery System (시비처리 방법에 따른 낙엽송 용기묘의 생장 및 생리 특성)

  • Cho, Min-Seok;Lee, Soo-Won;Park, Byung-Bae
    • Journal of Bio-Environment Control
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    • v.21 no.1
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    • pp.57-65
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    • 2012
  • Fertilization is essential to seedling production in nursery culture, but excessive fertilization can contaminate surface and ground water around the nursery. The objective of this study was to find optimal fertilization practice of container seedling production for reducing soil and water contamination around the nursery without compromising seedling quality. This study was conducted to investigate chemical properties of the growth medium, growth performance, chlorophyll fluorescence, and chlorophyll contents of larch ($Larix$ $kaempferi$) growing under three different fertilization treatments (Constant rate, Three stage rate, and Exponential rate fertilization). Root collar diameter and height of larch were not significantly different among treatments even though the nutrient supply of the exponential treatment was half that of the constant and three stage treatments. Chemical properties of the growth medium showed the same trends as root collar diameter and height. The total biomass and seedling quality index (SQI) were higher at Constant than at other treatments, but both SQI of Constant and Exponential were not significantly different. Photochemical efficiency and chlorophyll contents were lower at Exponential than at other treatments, but not significantly different among treatments. Therefore, Exponential fertilization which is 50% fertilizer of other treatments would maximize seedling growth and minimize nutrient loss.