• The Korean Journal of Physiology
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• v.30 no.1
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• pp.63-76
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• 1996

The aim of present study was to characterize phosphate uptake and to investigate the mechanism for the insulin and insulin-like growth factor(IGF) stimulation of phosphate uptake in primary cultured rabbit renal proximal tubule cells. Results were as follows : 1. The primary cultured proximal tubule cells had accumulated $6.68{\pm}0.70$ nmole phosphate/mg protein in the presence of 140 mM NaCl and $2.07{\pm}0.17$ nmole phosphate/mg protein in the presence of 140 mM KCl during a 60 minute uptake period. Raising the concentration of extracellular phosphate to 100 mM$(48.33{\pm}1.76\;pmole/mg\;protein/min)$ induced decrease in phosphate uptake compared with that in control cells maintained in 1 mM phosphate$(190.66{\pm}13.01\;pmole/mg\;protein/min)$. Optimal phosphate uptake was observed at pH 6.5 in the presence of 140 mM NaCl. Phosphate uptake at pH 7.2 and pH 7.9 decreased to $83.06{\pm}5.75%\;and\;74.61{\pm}3.29%$ of that of pH 6.5, respectively. 2. Phosphate uptake was inhibited by iodoacetic acid(IAA) or valinomycin treatment $(62.41{\pm}4.40%\;and\;12.80{\pm}1.64%\;of\;that\;of\;control,\;respectively)$. When IAA and valinomycin were added together, phosphate uptake was inhibited to $8.04{\pm}0.61%$ of that of control. Phosphate uptake by the primary proximal tubule cells was significantly reduced by ouabain treatment$(80.27{\pm}6.96%\;of\;that\;of\;control)$. Inhibition of protein and/or RNA synthesis by either cycloheximide or actinomycin D markedly attenuated phosphate uptake. 3. Extracellular CAMP and phorbol 12-myristate 13 acetate(PMA) decreased phosphate uptake in a dose-dependent manner in all experimental conditions. Treatment of cells with pertussis toxin or cholera toxin inhibited phosphate uptake. cAMP concentration between $10^{-6}\;M\;and\;10^{-4}\;M$ significantly inhibited phosphate uptake. Phosphate uptake was blocked to about 25% of that of control at 100 ng/ml PMA. 3-Isobutyl-1-methyl-xanthine(IBMX) inhibited phosphate uptake. However, in the presence of IBMX, the inhibitory effect of exogenous cAMP was not significantly potentiated. Forskolin decreased phosphate transport. Acetylsalicylic acid did not inhibit phosphate uptake. The 1,2-dioctanoyl-sn-glycorol(DAG) and 1-oleoyl-2-acetyl-sn- glycerol(OAG) showed a inhibitory effect. However, staurosporine had no effect on phosphate uptake. When PMA and staurosporine were treated together, inhibition of phosphate uptake was not observed. In conclusion, phosphate uptake is stimulated by high sodium and low phosphate and pH 6.5 in the culture medium. Membrane potential and intracellular energy levels are also an important factor fer phosphate transport. Insulin and IGF-I stimulate phosphate uptake through a mechanisms that involve do novo protein and/or RNA synthesis and decrease of intracellular cAMP level. Also protein kinase C(PKC) is may play a regulatory role in transducing the insulin and IGF-I signal for phosphate transport in primary cultured proximal tubule cells.

• The Korean Journal of Mycology
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• v.38 no.2
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• pp.152-159
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• 2010

The influence of urushiol, as an allergen on laccase property of Fomitella fraxinea was investigated. The enzyme production was reached to the highest level after 10 days, cultivation and the activity and mycelial biomass were increased by 2.5 and 1.5 folds, respectively, by adding urushiol in the culture medium. In liquid cultures using a Cu Mn-free medium, laccase lactivity was decreased by 3.8-9.2 folds, with similar dry cell weight. Two isoenzymes, were purified using anion exchange, hydrophobic interaction and size-exclusion chromatographies. Both isoenzymes are monomeric proteins, with $M_W$ around 67 kDa(Lac1) and 66 kDa(Lac2), and isoelectric points of 3.67 and 3.81. The optimal conditions for purified isoenzymes were found to be pH 4.5-5.0 and $30-35^{\circ}C$. Activity decreased by the addition of $Fe^{2+}$, $Mg^{2+}$, $Na^+$, and strongly inhibited by EDTA and sodium azide.

• Korean Journal of Plant Tissue Culture
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• v.28 no.2
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• pp.61-67
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• 2001

Using mature seed-derived callus, optimal conditions for efficient callus growth and plant regeneration, and regeneration efficiency by callus type were investigated in zoysiagrass (Zoysia japonica steud.). Callus induction was highest when the seeds were cultured on MS medium containing 2 mg/L 2,4-D, 0.2 mg/L BAP, 4 mg/L thiamine-HCl and 100 mg/L $\alpha$-ketoglutaric acid. Callus growth was highest when callus were cultured on MS medium containing 0.5 mg/L 2,4-D, 0.05 mg/L BAP, 4 mg/L thiamine-HCl and 100 mg/L $\alpha$-ketoglutaric acid. Plant regeneration was highest when callus was transferred on MS medium containing 3% maltose and 1 mg/L BAP, or 1 mg/L thidiazuron (TDZ). The combinations and concentrations of 2,4-D and BAP were shown to be critical factors for both the frequency and the type of callus. And four morphologically distinct types of callus were induced from the 2,4-D and BAP treatment. Type I,II and III calli produced shoots upon subculture, while the watery callus, type IV produced roots without shoots. Of four types of callus, type I exhibited the maximum frequency (82%) of shoot regeneration and the minimum frequency (4%) of albinism.

• Proceedings of the Korean Society of Life Science Conference
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• 2001.06a
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• pp.67-86
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• 2001

A strain producing strongly fibrinolytic enzyme was isolated from soil and was identified to be Bacillus subtilis by biochemical and physiological characterization. The optimal culture conditions for the production of fibrinolytic enzyme was determined to be 1.0% tryptone, 1.5% soluble starch, 0.5% Peptone, 0.5% NaCl, $(NH_{4})_{3}PO_4.3H_{2}O, and MgSO_{4}.7H_{2}O.$ Initial pH and temperature were pH 8.0 and $30^{\circ}C$ , respectively, The highest enzyme production was observed at 30 hours of cultivation at $30^{\circ}C$ The fibrinolytic enzyme was purified to homogeneity by DEAE Sephadex A-50 ion exchange column chromatography, 70% ammonium sulfate precipitation, Sephadex G-200 and G-75 gel filtration column chromatography. The molecular weight of the purified enzyme was 28,000 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A gene encoding the fibrinolytic enzyme was cloned into a plasmid vector pBluescript, transforming E.coli XL-1 Blue. The clone was able to degrade fibrin, This indicated that the gene could encode a fibrinolytic enzyme. The nucleotide sequence of the 2.7 kb insert was determined in both direction. One open reading frame composed of 1023 nucleotides was found to be a potential protein coding region. There was the putative Shine-Dalgano sequence and TATA box upstream of the open reading frame. The homology search data in the genome database showed that both the 2.7 kb insert and 1 kb open reading frame carried no significance in the nucleotide sequence of known fibrinolytic enzyme from Bacillus serovars. The recombinant cell harboring the novel gene involved in fibrinolysis was subjected to protein purification. The molecular mass of the purified fibrinolytic enzyme was determined to be 31864 Dalton, which was highly in accordance with the molecular mass(33 kDa) of the fibrinolytic gene deduced from the insert. The fibrinolytic enzyme was Purified 50.5 folds to homogeneity in overall yield of 10.7% by DEAE Sephadex A-50 ion exchange, 85% ammonium sulfate precipitation, Sephadex G-50, Superdex 75 HR FPLC gel filtration. In conclusion, a novel fibrinolytic gene from Bacillus subtilis was identified and characterized by cloning a genomic library of Bacillus subtilis into pBleuscript. For the soybean fermented by this strain, it is found that there increased assistant protein about 20% compared to the soybean not fermented and increased about 30% according to amino acid analysis and, in particular, essential amino acid increased about 40%. When keeping this fermented soybean powder at room temperature for about 70days, it showed very high stability maintaining almost perfect activity and, therefore, it gave us great suggestion its possibility of development as a new functional food.

• Korean Journal of Food Science and Technology
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• v.34 no.4
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• pp.661-665
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• 2002

In order to develop ginseng tea powder with spore forming lactic acid bacteria Lactobacillus sporogenes was used. In the jar fermentor experiment under optimal culture conditions, the number of spore of L. sporogenes reached about $20{\times}10^8\;CFU/mL$ and sporulation rate was 97%. Granulated ginseng tea was made from glucose 7 kg, lactose 2 kg, ginseng extract 1 kg and spores 5 g $(5200{\times}10^8\;CFU/g)$. In the treatment of artificial gastric juice (pH 3.0) for 4 h and artificial bile for 8 h, the survival rate of spores in the granulated ginseng tea was 55.4% and 90.0% respectively. The spores survived 77.6% after incubation for 20 min in boiled water. Its storage stability was about 75% for 12 months at room temperature.

• Korean Journal of Organic Agriculture
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• v.27 no.1
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• pp.65-76
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• 2019

Bacillus genus are found abundantly in various sites and their secondary metabolites were used as potential agents in agriculture, notably plant growth promoting and bio-control. The objective of this study was to develop the culture conditions of GH1-13 strain including higher cell growth, stable endospore-forming and enhancement of potential agents which are related with plant growth promoting and phytopathogen suppression. The optimal carbon and nitrogen sources were determined by glucose and soy bean flour, respectively, then resulted in $7.5{\times}10^9cells/mL$, $6.8{\times}10^9\;endospore\;cells/mL$ and sporulation yield of 90% after 30 h cultivation in 500 L submerged fermenter at $37^{\circ}C$, pH 7.0. Cells and cell-free supernatant of GH1-13 strains showed the potent antifungal activity against phytopathogenic fungi of Colletotrichum gloeosporioides. It was also confirmed that indole-3-acetic acid (IAA) production of GH1-13 strain was greatly increased by addition of 0.3% tryptophan.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2019.04a
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• pp.56-56
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• 2019

본 연구는 식용과 약용으로 이용되는 국내 자생식물인 야산고비(Onoclea sensibilis var. interrupta Maxim.)를 기내에서 대량증식하기 위한 조건을 구명하기 위하여 수행되었다. 무가온 온실에서 수집한 포자를 기내에서 발아시켜 전엽체를 획득하였으며, 계대배양하며 실험의 재료로 사용하였다. 전엽체의 대량증식을 위해 전엽체 0.3g을 메스로 균일하게 다진 후 증류수 1ml와 함께 배지에 고루 펼쳐서 배양하는 방법을 사용하였으며, 증식에 미치는 배지의 영향을 확인하기 위하여 1/4, 1/2, 1, 2배로 조절한 MS배지를 조성하여 8주간 배양하였다. 이후, 증식이 우수한 배지를 선정하여 sucrose와 질소급원의 농도를 조절하였으며, 활성탄을 첨가하여 증식에 미치는 영향을 확인하였다. 배지종류 실험의 결과, 생체중이 1MS에서 10.2g으로 초기 접종량에 비해 가장 많이 증가하였으며, 현미경을 이용하여 관찰한 결과에서도 정상적인 전엽체의 형태인 심장형으로 발달하였다. 증식이 우수하였던 1MS배지에 sucrose의 농도를 조절하여 배양한 결과에서는 1%의 처리구에서 가장 증식률이 좋았으며, 질소급원의 경우 30mM의 농도로 조절한 처리구가 가장 좋은 결과를 보였다. 배지 내 활성탄의 첨가는 처리구당 증가된 전엽체의 생체중이 유의적인 차이를 보이지 않았다. 포자체 대량 형성을 위한 적정 배양토의 혼합조건을 확인하기 위하여 원예상토, 피트모스, 펄라이트 및 마사토의 혼합비율을 5종류로 달리하여 조성하여 사각분($7.5{\times}7.5{\times}7.5cm$)에 충진하였다. 조성한 배양토에 기내에서 배양한 전엽체 1g을 증류수와 함께 블렌더를 이용하여 10초간 분쇄하여 토양표면에 고루 분주하였다. 이후 12주간 재배한 결과, 모든 토양조합에서 포자체가 형성되었다. 그중 원예상토와 마사토를 2:1(v:v)로 혼합한 토양에서 포트 당 405.0개로 가장 많은 포자체가 형성되었으며, 전체적인 생육 또한 비교적 양호한 결과를 보였다. 따라서 야산 고비의 전엽체 대량증식에 적합한 배지는 경제성과 생육수준을 고려하여 1%의 sucrose와 질소급원의 농도를 30mM로 조절한 1MS 배지가 적합하며, 포자체 대량생산을 위해서는 원예상토, 마사토를 2:1(v:v)로 혼합한 토양이 적합하다고 판단되었다.

• Korean Journal of Plant Resources
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• v.32 no.4
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• pp.290-295
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• 2019

This study was carried out to select the optimal condition for prothallus propagation and sporophyte formation of Leptogramma pozoi (Lag.) Ching subsp. mollissima (Fisch. ex Kunze) Nakaike and to provide basic data for mass production system. In the propagation, 300 mg of prothallus were inoculated in different kinds of medium [1/4, 1/2, Murashige and Skoog (MS), Knop], sucrose (0, 1, 2, 3, 4%), and cultured for 8 weeks. Sporophyte formation studies were carried out by inoculating blended prothallus into artificial soils. The soils were mixed with horticultural substrate, peatmoss, perlite, and decomposed granite at different ratios or only with the horticultural substrate. Then the prothallus was cultivated for 12 weeks to figure out the formation and growth of the sporophytes. The growth and development of prothalli were excellent on MS medium. According to medium components, prothallus growth was favorable in all treatments except 0% sucrose treatment and the highest in 2% sucrose. In the experiment of soil mixtures, sporophyte formation was the highest in the horticultural substrate:perlite 2:1 (v:v) and horticultural substrate:decomposed granite 2:1 (v:v) treatment, and the overall growth was good in the horticultural substrate:decomposed granite 2:1 (v:v) mixed soil.

• Journal of Microbiology and Biotechnology
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• v.32 no.5
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• pp.630-637
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• 2022

The objective of this study was to optimize industrial-grade media for improving the biomass production of Weissella cibaria JW15 (JW15) using a statistical approach. Eleven variables comprising three carbon sources (glucose, fructose, and sucrose), three nitrogen sources (protease peptone, yeast extract, and soy peptone), and five mineral sources (K₂HPO₄, potassium citrate, ⳑ-cysteine phosphate, MgSO₄, and MnSO₄) were screened by using the Plackett-Burman design. Consequently, glucose, sucrose, and soy peptone were used as significant variables in response surface methodology (RSM). The composition of the optimal medium (OM) was 22.35 g/l glucose, 15.57 g/l sucrose, and 10.05 g/l soy peptone, 2.0 g/l K₂HPO₄, 5.0 g/l sodium acetate, 0.1 g/l MgSO₄·7H₂O, 0.05 g/l MnSO₄·H₂O, and 1.0 g/l Tween 80. The OM significantly improved the biomass production of JW15 over an established commercial medium (MRS). After fermenting OM, the dry cell weight of JW15 was 4.89 g/l, which was comparable to the predicted value (4.77 g/l), and 1.67 times higher than that of the MRS medium (3.02 g/l). Correspondingly, JW15 showed a rapid and increased production of lactic and acetic acid in the OM. To perform a scale-up validation, batch fermentation was executed in a 5-l bioreactor at 37℃ with or without a pH control at 6.0 ± 0.1. The biomass production of JW15 significantly improved (1.98 times higher) under the pH control, and the cost of OM was reduced by two-thirds compared to that in the MRS medium. In conclusion, OM may be utilized for mass producing JW15 for industrial use.

• Korean Journal of Plant Resources
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• v.35 no.4
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• pp.502-507
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• 2022

In the present study, plant regeneration through adventitious shoot formation from hypocotyl segments of in vitro seedlings of groundcherry (Physalis angulata L.) was investigated to determine the optimum culture conditions for highly efficient regeneration of the species. Adventitious shoots in hypocotyl segments were efficiently induced on MS media with low concentrations of BAP, specifically, with 0.5-1.0 mg/L BAP singly or in combination with 0.1-0.5 mg/L NAA. The 1.0 mg/L BAP single treatment was most effective for forming multiple adventitious shoots. When the induced shoots were transferred to the root induction media, low concentrations of NAA, IBA, and IAA enhanced the development of adventitious roots from adventitious shoots, suggesting that low concentrations of auxins were optimal for producing regenerated plantlets. The number of roots per shoot was large (> 2.0), and the root length exceeded 8.0 cm. In particular, the development and the overall shape of the roots were ideal. Furthermore, the number and length of shoots exceeded 2 and 6.0 cm, respectively. When the regenerated plantlets were transferred to compost soil, the root and shoot systems had developed well to the point that all of the regenerated plantlets acclimated successfully, resulting in normal morphology and growth characteristics, similar to those of the mother plant. Therefore, plant regeneration via adventitious shoot formation is expected to be one of the main methods for producing groundcherry on a large scale for a stable supply of the raw materials.