• Mycobiology
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• v.41 no.3
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• pp.159-163
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• 2013

A chemical mutagenesis technique was employed for development of mutant strains of Sparassis crispa targeting the shortened cultivation time and the high ${\beta}$-glucan content. The homogenized mycelial fragments of S. crispa IUM4010 strain were treated with 0.2 vol% methyl methanesulfonate, an alkylating agent, yielding 199 mutant strains. Subsequent screening in terms of growth and ${\beta}$-glucan content yielded two mutant strains, B4 and S7. Both mutants exhibited a significant increase in ${\beta}$-glucan productivity by producing 0.254 and 0.236 mg soluble ${\beta}$-glucan/mg dry cell weight for the B4 and S7 strains, respectively, whereas the wild type strain produced 0.102 mg soluble ${\beta}$-glucan/mg dry cell weight. The results demonstrate the usefulness of chemical mutagenesis for generation of mutant mushroom strains.

• Microbiology and Biotechnology Letters
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• v.23 no.5
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• pp.588-592
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• 1995

Fed-batch fermentation was used to produce the high concentrations of poly-$\beta $-hydroxybutyrate (PHB) and poly-$\beta $-(hydroxybutyrate-co-hydroxyvalerate) (PHB/V). Specific growth rate ($\mu $), yield of cell from glucose (Y$_{x/s}$) were calculated from the two samples in 3 to 5 hours of interval and they were reflected on the determination of glucose feeding rate to maintain the glucose concentration at around 10 g/l in the culture broth. PHB was accumulated after the nitrogen became limited at 60 g/l of dry cell weight by changing ammonia water to 4N-NaOH solution. As results, the final dry cell weight (DCW) of 170 g/l, PHB of 115 g/l were obtained in 50 hours and the overall productivity was 2.4 g/l$\cdot $h. After PHB accumulation, cosubstrate of glucose and propionic acid (PA) was fed to accumulate PHB/V. But, PA feeding rate was decreased from 3 g/l$\cdot $h to 1 g/l$\cdot $h to prevent PA from accumulating to high level in the broth, which is very inhibitory to the cells. As results, DCW, PHB and PHV were 147.5 g/l, 90 g/l and 8 mole % of hydroxyvalerate, respectively.

• Journal of Microbiology and Biotechnology
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• v.9 no.5
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• pp.593-603
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• 1999

Poly(3-hydroxybutyrate) (PHB) production by fermentation was examined under both restricted- and ample-oxygen supply conditions in a single fed-batch fermentation. Recombinant Escherichia coli transformed with the PHB production plasmid pSYLl07 was grown to reach high cell density (227 g/l dry cell weight) with a high PHB content (78％ of dry cell weight), using a glucose-based minimal medium. A simple flux model containing 12 fluxes was developed and applied to the fermentation data. A superior closure (95％) of the carbon mass balance was achieved. When the data were put into use, the results demonstrated a surprisingly large excretion of formate and lactate. Even though periods of severe oxygen limitation coincided with rapid acetate and lactate excretion, PHB productivity and carbon utilization efficiency were not significantly impaired. These results are very positive in reducing oxygen demand in an industrial PHA fermentation without sacrificing its PHA productivity, thereby reducing overall production costs.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.3
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• pp.192-197
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• 2005

The effects of macronutrients $(NO_3^-,\; NH_4^+\;and\;PO_4^{3-})$ on cell growth and triterpenoids production in Centella asiatica cell suspension cultures were analyzed using the Box­Behnken response surface model experimental design. In screening and optimization experiments, $PO_4^{3-}$ as a single factor significantly influenced cell growth where increasing the phosphate level from 0.1 to 2.4 or 2.6 mM, elevated cell growth from 3.9 to $14\~16g/L$. The optimum values predicted from the response surface model are 5.05mM $NH_4^+$, 15.0mM $NO_3^-$ and 2.6mM $PO_4^{3-}$, yielding 16.0g/L cell dry weight with $99\%$ fitness to the experimental data. While the $NH_4^+-NO_3^-$ interaction influenced cell growth positively in the optimization experiment, $NH_4^+$ and $NO_3^-$ as single factors; and interactions of $NO_3^--PO_4^{3-},\;NH_4^+-PO_4^{3-}$ and $NH_4^+-NO_3^-$ were all negative in the screening experiment. Cell growth and the final pH level were positively affected by $PO_4^{3-}$, but negatively affected by $NH_4^+\;and\;NH_4^+-PO_4^{3-}$ interactions. The different effects of factors and their interactions on cell growth and final pH are influenced by a broad or narrow range of macronutrient concentrations. The productions of triterpenoids however were lower than 4mg/g cell dry weight.

• KSBB Journal
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• v.11 no.2
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• pp.201-210
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• 1996

The production of polyhydroxyalkanoate(PHA) from glucose by batch and fed-batch culture of Pseudomonas sp. HJ was studied. In batch culture using fermentor, 400 rpm of agitalion speed, 2 vvm of aeration rate, 18 hours of inoculum age, and 5% (vlv) of inoculum size were optimal. PHA production was not increased by deficiency of oxygen. In a batch culture, the final call mass was $6.251g/\ell$, and PHA content was 20% of dry cell weight. In a constant feeding fed-batch culture, cell mass increased to $33.24g/\ell$, and PHA content reached 48.9% of dry cell weight. In an intermittent feeding fed-batch culture, cell mass increased to $37.89g/\ell$, and PHA content reached 53.5% of dty cell weight.

• Journal of Plant Biotechnology
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• v.45 no.4
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• pp.340-346
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• 2018

In this study, the effect of elicitors such as yeast extract (YE), methyl jasmonate (MeJA) and salicylic acid (SA) on the accumulation of eurycomanone in Eurycoma longifolia cell cultures were investigated. Suspension cells of E. longifolia was cultured in Murashige and Skoog (MS) medium supplemented with 30 g/L sucrose, 1.25 mg/L naphthaleneacetic acid (NAA) and 1 mg/L kinetin at a shaking speed of 120 rpm. Elicitors were added in the culture at different concentrations and times to stimulate eurycomanone accumulation in the Eurycoma longifolia cells. Eurycomanone content was determined by HPLC with a C18 column, flow rate of 0.8 mL/min, run time of 17.5 min, and a detector wavelength of 254 nm. The stationary phase was silica gel and the mobile phase was acetonitrile: $H_2O$. Non-elicited cells were used as the control. The study showed the effect of different elicitor concentrations, YE at 200 mg/L, MeJA at $20{\mu}M$ and SA at $20{\mu}M$ stimulated high production of eurycomanone. In which, treatment of $20{\mu}M$ MeJA after 4 days of culture resulted in the highest accumulation of this compound (17.36 mg/g dry weight), approximately 10-fold higher than that of untreated cells (1.70 mg/g dry weight).

• KSBB Journal
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• v.11 no.4
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• pp.398-404
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• 1996

Escherichia coli was inoculated in calcium alginate capsules and cultivated to prepare encapsulated whole cell ${\beta}$-galactosidase. The dry cell weight in the capsule reached 100 g/L based on the inner space of the capsule. The activity of the encapsulated whole cell ${\beta}$-galactosidase increased with the dry cell weight increase during cultivation in the production medium. The activity of the encapsulated whole cell ${\beta}$-galactosidase was increased 25% by adding $2{\times}10^{-4}M Zn^{+2}$ ion in the production medium and 10% by coencapsulating with 2% (v/v) sunflower seed oil. The activity of encapsulated whole cell ${\beta}$-galactosidase produced in the concentric air lift reactor in which kLa was 82/hr was 86% higher than that in the shaking flask incubator where kLa was 2.55/hr.

• Journal of Plant Biology
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• v.28 no.2
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• pp.141-148
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• 1985

The effects of sizes and densities of cells cultured, conditioned medium, and media pH on the somatic embryogenesis of carrot (Daucus carota L.) were examined. A large number of globular embryoids was formed after 4 days in cell culture, and later globular embryoids developed into heart and torpedo shape. High cell density resulted in higher number and better growth of embryos, especially on conditioned medium than Murashige-Skoog medium. The fresh weight and number of embryoids formed increased with the decrease in cell size. The significant reduction in fresh weight and number of embryoids was obtained when culturing cells with diameter of over 90 ${\mu}{\textrm}{m}$. Dry weight and number of embryoids were markedly reduced with medium pH of 4 or 7, but promoted with pH 6.0.

• Korean Journal of Plant Tissue Culture
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• v.24 no.6
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• pp.361-367
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• 1997

This study was carried out to select cadmium resistant cell lines from leaf-derived calli of diploid and haploid of Nicotiana tabacum cv. BY4, for understanding adaptation mechanism of plants in cadmium contaminated environment. suspended cell clumps were plated onto selection medium containing 0 to 2,000 $\mu$M cadmium. Cadmium resistant colonies were formed on the selection medium after 3 or 4 weeks of culture. The minimum inhibition concentration (MIC) of cadmium on colony formation were 300 $\mu$M in diploid and 200 $\mu$M in haploid plants, respectively. In order to test the resistance to cadmium, selected cell line on MIC were transferred to medium containing high concentration of cadmium. The selected cell lines, especially haploid cell line, were resistant an the high concentration of cadmium. And dry weight, ash weight, and cadmium contents of cell were increased. These results indicated that the selected cell lines showed higher resistance of cadmium than control cells, and haploid plant is more resistant than diploid plant on medium with cadmium.

• KSBB Journal
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• v.13 no.5
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• pp.569-576
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• 1998

The effects of carbohydrates, hormones and elicitors on both cell growth and betaine production were investigated in the cell cultures of Lycium chinense Mill. The maximum effect of glucose and sucrose was observed in cells cultured in the presence of 3% and 7% for cell growth and betaine production, respectively. The effect of hormones on cell growth and betaine production was prominent in the presence of 10 ${\mu}$M 2, 4-D, 10 ${\mu}$M NAA and 2.5 ${\mu}$M IAA, whereas cell growth and betaine production were excellent at 2.5 ${\mu}$M BA and 10 ${\mu}$M BA, respectively. Abiotic elicitors such as KCI, MnCl2 and NaCl exhibited an inhibitory role on cell growth in all treatment groups. Betaine production was increased according to increase of concentration of abiotic elicitors. methanol-soluble and insoluble components as biotic elicitor remarkably inhibited cell growth from 2 mg and 6 mg, respectively. Betaine production was increased maximally at 2 mg of biotic elicitors. When growth medium was switched to production medium at two stage culture, it resulted that cell fresh weight and dry weight decreased but betaine content increased about 2.2-fold.