• Journal of Plant Biotechnology
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• v.46 no.2
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• pp.127-135
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• 2019

The purpose of this study was to investigate suitable parts for callus induction and optimal concentrations of growth regulators, contained in the medium affecting shoot and rooting for in vitro mass production of Haworthia truncata. Leaves and flower bud showed 100% callus formation rate at NAA $1{\sim}2mgL^{-1}$ treatment, and NAA $1mgL^{-1}$ + TDZ $2mgL^{-1}$ treatment. The flower stalk showed 75% callus formation rate, at NAA $2mgL^{-1}$ + TDZ $2mgL^{-1}$ treatment in H. truncata. While the rate of callus formation was high in leaves and flower bud, leaves were the most efficient in obtaining most culture parts. Shoot induction rate from callus was highest, at NAA $0.1mgL^{-1}$ treatment in H. truncata. Additionally, the number of shoots formation was 66.3 shoots high, in NAA $1mgL^{-1}$ + BA $0.1mgL^{-1}$ treatment in H. truncata. In the case of acclimatization of regenerated plant, growth characteristics did not show significant difference (95%) shading with respect to the different ratio of substrate mixture, and it was determined that would be appropriate, considering plant height and appearance preference of H. truncata. It was established that optimization of culture condition, was responsible for mass propagation in vitro cultures of H. truncata.

• Journal of Plant Biotechnology
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• v.46 no.1
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• pp.22-31
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• 2019

The objective of this study was to investigate the suitable parts for callus induction and optimal concentrations of growth regulators contained in the medium affecting shooting and rooting Echeveria laui and Echeveria elegans for in vitro mass production. To determine the suitable plant parts for callus induction, the leaves were divided into upper, medium and bottom parts and cultured on MS medium at different concentrations with $0{\sim}2mgL^{-1}\;NAA$ and $0{\sim}4 mgL^{-1}BA$. The upper and middle parts of leaves both showed 100% callus formation rate with $NAA\;1\;mgL^{-1}$ and $BA\;1\;mgL^{-1}$ treatment in E. laui. The middle parts of leaves showed 83.3% callus formation rate at $NAA\;2\;mgL^{-1}$ and BA 4 mgL-1 treatment in E. elegans. The shoot induction rate from callus was highest at $NAA\;0.1\;mgL^{-1}$ and $BA\;3\;mgL^{-1}$ treatment in E. laui and $NAA\;0.3\;mgL^{-1}$ in E. elegans. In addition, the number of shoots formation was 10.4 shoots high in $NAA\;1\;mgL^{-1}$ and $BA\;1\;mgL^{-1}$ treatment in E. laui and 12.0 shoots in most effective $NAA\;1\;mgL^{-1}$ and $BA\;0.1\;mgL^{-1}$ treatment in E. elegans. In the case of acclimatization of regenerated plant, growth characteristics did not show any significant difference (35 ~ 55%) shading with respect to the different ratio of substrate mixture, and it was determined that would be appropriate considered plant height and appearance preference of E. laui and E. elegans. It was established that the optimization of culture condition was responsible for the mass propagation in vitro cultures of E. laui and E. elegans.

• Journal of Life Science
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• v.31 no.1
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• pp.90-99
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• 2021

In this study, to reduce the production cost of poly(3-hydroxyalkanoates) (PHA), optimal cell growth and PHA biosynthesis conditions of the isolated strain Pseudomonas sp. EML8 were established using waste frying oil (WFO) as the cheap carbon source. Gas chromatography (GC) and GC mass spectrometry analysis of the medium-chain-length PHA (mcl-PHAWFO) obtained by Pseudomonas sp. EML8 of WFO indicated that it was composed of 7.28 mol% 3-hydrxoyhexanoate, 39.04 mol% 3-hydroxyoctanoate, 37.11 mol% 3-hydroxydecanoate, and 16.58 mol% 3-hydroxvdodecanoate monomers. When Pseudomonas sp. EML8 were culture in flask, the maximum dry cell weight (DCW) and the mcl-PHAWFO yield (g/l) were showed under WFO (20 g/l), (NH4)2SO4 (0.5 g/l), pH 7, and 25℃ culture conditions. Based on this, the highest DCW, mcl-PHAWFO content, and mcl-PHAWFO yield from 3-l-jar fermentation was obtained after 48 hr. Similar results were obtained using 20 g/l of fresh frying oil (FFO) as a control carbon source. In this case, the DCW, the mcl-PHAFFO content, and the mcl-PHAFFO yields were 2.7 g/l, 62 wt%, and 1.6 g/l, respectively. Gel permeation chromatography analysis confirmed the average molecular weight of the mcl-PHAWFO and mcl-PHAFFO to be between 165-175 kDa. Thermogravimetric analysis showed decomposition temperature values of 260℃ and 274.7℃ for mcl-PHAWFO and mcl-PHAFFO, respectively. In conclusion, Pseudomonas sp. EML8 and WFO could be suggested as a new candidate and substrate for the industrial production of PHA.

• Journal of Bio-Environment Control
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• v.32 no.3
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• pp.249-255
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• 2023

Korean melon (Cucumis melo L.) is grown mostly in Northeast Asia area, and as a fruit mainly produced in Korea, the yield per unit area continues to improve, but the cultivation method is limited to soil cultivation, so it is necessary to develop hydroponic cultivation technology for scale and labor-saving is needed. As the ratio of NO₃^- increased, the plant height, the leaf length, the leaf width, and the internode length became longer and larger. On the other hand, the SPAD value decreased. The lower the ratio of NO₃^-, the faster the female flower bloom, and there was no difference in fruit maturity between treatments. There was no difference in the shape of fruit according to the ratio of NO₃^-, and the hardness was higher as the ratio of NO₃^- was lower. The total yield from March to July was KM3 5,650 kg/10a and KM1 4,439 kg/10a, 27% higher in KM3 and, in particular, 36% higher in quantity from March to May, when Korean melon prices were high season. Therefore, it was judged that it would be appropriate to supply NO₃^- suitable for hydroponic cultivation of Korean melon, which was formalized in December and produced from spring, at the level of 6.5 to 10 me·L^-1.

• Applied Biological Chemistry
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• v.27
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• pp.29-46
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• 1984

To utilize several species of hard wood as raw materials of feed products, fermentation characteristics of cellulosic substrates to single cell protein was investigated, and results were summarized as follows. Among the microorganisms investigated, Tricoderma viride was selected as one of the most cellulolytic. Mixed culture of fungi did not show a synergistic effect on cellulose degradation. When the fungi were cultured at $28^{\circ}C$ for 7 days in a medium containing wheat bran 25 g, cellulose 0.25 g, proteose peptone 0.025 g and tween 800.025 g, cellulotic activities on carboxy methyl cellulose and filter paper reached maximum at 12 hr. The alkali treatment resulted in increased degradation of substrate from 13 to 18% when treated with enzymes for 12h, and reducing sugar formation increased with decreased size of substrates. Glucose was a very good feedback inhibitor of the enzyme from T.viride than that of xylose. When the substrate was rehydrolyzed, hydrolysis rate was 31% to reducing sugars within 12 hr. Quantative anlysis with HPLC showed the ratio of glucose to xylose in sugar syrups as 1.77 to 1. For the purpose of producing cellulosic-single cell protein from the sawdust of mulberry tree, 15 strains of xylose-assimilating yeast were isolated from 42 samples of rotten woods and compost soils and examined for their ability to utilize xylose. Then three strains were selected by their strong xylose-assimilating activities. The cultivative condition, the growth characteristics, and protein and nucleic acid productivities of three strains were investigated. The results obtained were, 1. Wood hydrolysate of mulberry tree was assimilated by 5 strains of CHS-2, CHS-3, ST-40, CHS-12 and CHS-13. 2. The optimum initial pH and temperature for the growth of strain CHS-13 were 4.4 and $30^{\circ}C$. 3. The specific growth rate of strain CHS-13 was $0.23h^{-1}$ and generation time was 3.01 hrs at the optimum condition. 4. CHS-13 strain assimilated 81 % of sugar in wood hydrolysate. 5. CHS-13 strain was identified as Candida guilliermondii var. guilliermondii 6. When the CHS-13 strain was cultured in the wood hydrolysate containing yeast extract, L-protein content was increased with yeast extract concentration. 7. The L-protein and nucleic acid yields from wood hydrolysate were 0.73 mg/ml and $4.92{\times}10^{-2}\;mg/ml$ respectively. 8. An optimal nucleic acid content of CHS-13 strain was observed in the medium containing 0.2% of yeast extract.

• Journal of Mushroom
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• v.13 no.1
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• pp.56-62
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• 2015

A lipase producing bacterium was isolated from button mushroom bed, which showing high clear zone on agar media containing Tributyrin as the substrate. The strain was identified as Burkholderia cepacia by analysis of 16S rDNA gene sequence. Crude lipase (CL) was partially purified from 70% ammonium sulfate precipitation using the culture filtrate of B. cepacia. Immobilized lipases were prepared by cross-linking method with CL from B. cepacia and Novozyme lipase (NL) onto silanized Silica-gel as support. Residual activitiy of the immobilized CL (ICL) and immobilized NL (INL) was maintained upto 61% and 72%, respectively. Biodiesel (Fatty acid methyl ester, FAME) was recovered by transesterification and methanolysis of Canola oil using NaOH, CL and ICL as the catalysts to compare the composition of fatty acids and the yield of FAME. Total FAME content was NaOH $781mg\;L^{-1}$, CL $681mg\;L^{-1}$ and ICL $596mg\;L^{-1}$, in which the highest levels of FAME was observed to 50% oleic acid (C18:1) and 22% stearic acid (C18:0). In addition, the unsaturated FAME (C18:1, C18:2) decreased, while saturated FAME (C16:0, C18:0) increased according to increasing the reaction times with both CL and ICL, supporting CL possess both transesterification and interesterification activity. When reusability of ICL and INL was estimated by using the continuous reaction of 4 cycles, the activity of ICL and INL was respectively maintained 66% and 79% until the fourth reaction.

• Microbiology and Biotechnology Letters
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• v.34 no.1
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• pp.7-14
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• 2006

Feather, generated in large quantities as a byproduct of commercial poultry processing, is almost pure keratin, which is not easily degradable by common professes. Four strains, SMMJ-2, FL-3, NO-4 and RM-12 were isolated from soil for production of extracellular keratinolytic protease. They were identified as Bacillus sp. based on their morphological and physiological characteristics. They shown high protease activity on 5.0% skim milk agar medium and produced a substrate like mucoid on keratin agar medium. Bacillus sp. SMMJ-2 had a faster production time for producing keratinolytic protease than other strains. This strain did not completely degrade whole chicken feather for five days in basal medium but completely degraded whole chicken feather when supplied with nitrogen source for 40hours in keratinolytic producing medium ($0.7%\;K_{2}HPO_{4},\;0.2%\;KH_{2}PO_{4},\;0.1%$ fructose, 1.2% whole chicken feather, $0.01%\;Na_{2}CO_3$, pH 7.0). When supplied with chicken feather as nitrogen source, keratinolytic protease activity was 89 units/ml/min. When soybean meal was used as nitrogen source, the keratinolytic protease production reached a maximum of 106 units/ml/min after 48 hours under $30^{\circ}C$, 180 agitation. To isolate the keratinolytic protease, the culture filtrate was precipitated with $(NH_4)_{2}SO_4$ and acetone. The recovery rate of keratinolytic protease was about 96% after treatment with 50% acetone. The enzyme was stable in the range of $30{\sim}50^{\circ}C$ and pH $6.0{\sim}12.0$.

• The Korean Journal of Pharmacology
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• v.26 no.1
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• pp.1-6
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• 1990

We established an in vitro system of central serotonergic neurons by culturing dissociated rat embryonic (El4) brainstem cells to 14 days in vitro and monitored the serotonergic neuronal growth by measuring 5-hydroxytryptamine (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) contents in the cells with hish performance liquid chromatography with electrochemical detection (HPLC-EC). We studied also tile effects of various drugs on the contents of 5-HT and 5-HIAA, confirming in vivo reports. The 5-HT content (13 ng/mg protein) and 5-HT turnover rate (17 pmol/mg protein/h) at 14 days in vitro were in good agreement with those reported in the adult rat brain. The 5-HT content was more easily depleted with p-chlorophenylalanine, a tryptophan hydroxylase inhibitor than with NSD 1015 (3-hydroxybenzylhydrazine), an aromatic L-amino acid decarboxylase (AADC) inhibitor. Incubation of the cultures with tryptophan or 5-hydroxytryptophan increased the rate of serotonin formation implying that neither tryptophan hydroxylase nor AADC is saturated with its amino acid substrate in this in vitro system . The 5-HT content was depleted by reserpine. The 5-HT and 5-HIAA contents were increased and decreased, respectively, by monoamine oxidase inhibitors. All the above results indicate that the biochemical properties of the central serotonergic neurons in this culture system reflect reliably those of central serotonergic neurons in vivo. We suggest that measuring 5-HT and 5-HIAA contents in the primary cultured dissociated brainstem-cells with HPLC-EC is useful in the study of pharmacology as well as toxicolgy of the central serotonergic neurons.

• The Korean Journal of Mycology
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• v.26 no.2 s.85
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• pp.265-274
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• 1998

This study was carried out to obtain the basic data for an artificial cultivation of Phellinus sp.. The optimum conditions for the mycelial growth on the different sawdusts (Quercus aliena, Morns alba and Alnus japonica) substrate of an isolated Phellinus sp. were optimized by response surface methodology. The ratio of rice bran addition to sawdust and the suitable moisture content for the mycelial growth in the all sawdust media were about 30% (w/w) and $65{\sim}70%$ (w/v), respectively. The initial pHs for the mycelial growth of Quercus aliena and Morns alba were in the range of $pH\;5{\sim}6$, whereas Alnus japonica was obtained at pH 6. The optimum temperature for the mycelial growth was about $25{\sim}30^{\circ}C$, depending on the different kinds of wood substrates. From the response surface analysis, the values of independent variables of Quercus aliena at stationary points were determined to be 31.01 % (w/w) of rice bran, pH of 5.31 and 69.03% (w/v) of moisture content, and the expected value of mycelial growth was about 8.32 cm. Both the ratio of rice bran addition to sawdust $(X_1)$ and moisture content $(X_3)$ were effective to the mycelial growth. In the case of Morns alba, the ratio of rice bran addition to sawdust, initial pH and moisture content at the stationary points were 28.77% (w/w), 5.28 and 69.8 (w/v),respectively, and the expected mycelial growth of 7.60 cm was obtained. Stationary points for the mycelial growth in the sawdust media of Alnus japonica were 28.74% (w/w) of rice bran, pH of 6. 04 and 66.96% (w/v) of moisture content, and the expected values of mycelial growth was about 5.38 cm. Based on the above results, there was correlations between the mycelial growth and independent variables, and the effect of rice bran $(X_1)$ and initial pH $(X_2)$ for the mycelial growth were higher than the moisture content $(X_3)$. The optimum species of sawdust media for the my celial growth of Phellinus sp. was in the order of Quercus aliena > Morns alba > Alnus japonica.

• THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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• v.6 no.1
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• pp.81-97
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• 2000

Objectives: This experimental study was carried out to evaluate the effects of aqueous and methanol extracts of Hedyotis diffusa which has long been used for cancer treatment in oriental medicines on the induction of apoptotic cell death in human lymphoid leukemia cell line, HL-60. Methods: Cells were treated with various concentrations (200 to $0.4{\mu}g$) and periods (6 to 30 hr) of $H_2O$ and methanol extracts of Hedyotis diffusa. Then, cells were tested for viability by MTT assay. Cells wrere treated with $200{\mu}g/ml$ of methanol extract fork various periods. Genomic DNA was isolated, separated, on 1.5% agarose gels, stained with ethidium bromide and visualized under UV light. Cells were treated with $200{\mu}g/ml$ of each extract for 16 hr. Then, cells were treated with Hoechst dye 33342 and observed by fluorescence microscopy. Cells were treated with various doses of each for 12 hr and $100{\mu}g/ml$ of methanol extract for various periods. Lysate from the cells used to measure the activity of Caspase-1 and-3 proteases by using fluorogenic peptide substrates including acetyl-YVAD-AMC and acetyl-DEVD-AMC, respectively. Cells were treated with $200{\mu}g/ml$ of each extract for various periods. Cell lysates were immunoprecipated with anti-JNKl antibodies. The immune complex was reacted with $32^p-ATP$ and c-Jun as a substrate. The phosphotransferase activity of JNKI was measured by using PhosphoImage analyzer (Fuji Co., Japan). Nuclear extracts were isolated and incubated with oligonucleotide probe of $NF-{\kappa}B$. Transcriptional activation of ${\kappa}B$ was measured by using EMSA and visualized by PhosphoImage analyzer (Fuji Co, Japan). Cell lysates were prepared and analyzed by Western blotting with anti-Bc12 antibodies and anti-Bax antibodies. Cells were pretreated with various doses of methanol extract for 2 hr. Then, the extract was removed by centrifugation. Cells were resuspended with RPMI-1640 media containing 0.3% agarose, 10% FBS, overlayred onto bottom layer agarose and incubated at $CO_2$ incubator for 6 days. The number of colony was counted under light microscopy ($\time100$). Results: The death of HL-60 cells was markedly induced by the addition of methanol extract of Hedyotis diffusa in a dose and time-dependent manners. The apoptotic characteristic ladder pattern of DNA strand break was observed in death of HL-60 cells. In addition, it was shown nucleus chromatin condensation and fragmentation under Hoechst staining. Therefore, Hedyotis diffusa extract-induced death of HL-60 cells is mediated by apoptotic signaling processes. The activity of Caspase 3-like proteases remained in a basal level in HL-60 cells treated with aqueous extract of Hedyotis diffusa. However, it was markedly increased in HL-60 cells treated with methanol extract of Hedyotis diffusa. In addition, the phosphotransferase activity of JNKl was increased in HL-60 cells treated with methanol extract of Hedyotis diffusa. Furthermore, the activation of transcriptional activator, $NF-{\kappa}B$ was markedly induced by methanol extract of Hedyotis diffusa. Anti-apoptotic Bc12 was cleaved into 23Kda fragment by treatment of methanol extract of Hedyotis diffusa. However, expression of proapoptotic Bax protein was increased by treatment of methanol extract of Hedyotis diffusa in a time-dependent manner. Furthermore, methanol extract markedly inhibited the colony forming efficiency of HL-60 cells in semisolid agar culture. Conclusions: Above results suggest that methanol extract of Hedyotis diffusa induces the apoptotic death of human leukemic HL-60 cells via activations of Caspase-3 proteases, JNKI, transcriptional activator $NF-{\kappa}B$, In addition, our results also suggest that methanol extract of Hedyotis diffusa reduces the malignant potential of HL-60 cells via down regulation of colony forming effciency through cleavage of Bc12 as well as induction of Bax.