• Mycobiology
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• 제41권3호
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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.

• 한국미생물·생명공학회지
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• 제23권5호
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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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• 제9권5호
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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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• 제10권3호
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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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• 제11권2호
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• pp.201-210
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• 1996

본 연구는 단일 탄소원인 포도당으로부터 PHA를 생산하는 공시균 Pseudomonas sp. HJ의 유가배양 을 검토함으로써, PHA 대량생산을 위한 기초자료 를 얻는데 그 목적을 두었으며 그 결과는 다음과 같다. U 발효조를 이용하여 배양시 교반속도 4 400rpm, 통기량 2vvm, 종균 배양시간 18 시간 및 종균 접종량 5%(v/v)일때 최적 생육을 나타내었다. 공시균 Pseudomonas sp. HJ는 용존산소의 결핍 에 의해서 PHA 축적율이 향상되지 않았으며, P PHA 생산단계에 충분한 수준으로 용존산소를 유지 시켜 주어야 함을 알 수 있었다. 공시균 Pseudomo­7 nas sp. HJ를 회분배양했을 때 24시간만에 $6.251g/\ell$의 건조균체량과 20% 의 PHA 축적율을 나타내었으며, constant feeding fed-batch culture로 배 양 45시간만에 최고$33.24g/\ell$의 건조균체량과 48.9%의 PHA 축적율을 나타내었다. 또한 intermit­t tent feeding fed-batch culture로 공시균 Pseu domonas sp. HJ를 배양했을때, 배양 45시간만에 $37.89g/\ell$의 건조균체량과 53.5%의 PHA 축적율을 나타내였다.

• Journal of Plant Biotechnology
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• 제45권4호
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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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• 제11권4호
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• pp.398-404
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• 1996

전세포 효소 고정화 캡슐을 제조하기 위하여 Eschericia coli를 calcium alginate 캡슐내부에 접종하고 배양하였다. E. coli의 캡슐내부 건조중량이 1OOg/L에 달하였다. 생산배지에서 배양하는 동안 캡슐내부에 축적되는 미생물의 양이 많을수록 캡슐 내부에 축적되는 $\beta$galactosidase의 활성도 높았다. 생산배지에 금속이온 $Zn^{+2}를 2{\times}10^{-4} M $ 첨가함으로써 캡슐내부에 축척되는 ${\beta}$-galactosidase의 활성 을 25% 증가시킬 수 있었다. 캡슐제조시 해바라기유를 부피비로 2% 첨가함으로써 캡슐내부에 축적되 는 ${\beta}$-galactosidase의 활성을 10% 증가시킬 수 었었다. 부피산소전달계수, $k_La가 2.55h^- $ 안 플라스크 대신 $k_La가 82h^- $인 concentric air lift reactor 내에서 고정화 E. coli를 배양호농로써 캡슐내부의 전세포${\beta}$-galactosidase의 활성을 86% 증가시킬 수 있었다.

• Journal of Plant Biology
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• 제28권2호
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• pp.141-148
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• 1985

당근(Daucus carota L.) 세포 배양시 무성배 발생에 미치는 세포의 크기, 세포의 밀도, conditioned medium및 배지의 pH효과를 구명하기 위하여 실험한 결과는 다음과 같다. 세포배양시 구상형 배의 발생은 배양 4일째 최대에 달했으며 그 이후부터는 심장형 및 어뢰형 배로 발달되었다. 세포의 밀도가 높을수록 배 발생수가 증가하였으며 MS배지보다는 conditioned medium에서 양호하였고 배의 발달도 촉진되었다. 세포의 크기가 작을수록 생체중 및 배의 형성수가 증가하였으며 9o$\mu\textrm{m}$이상의 세포에서는 현저히 억제되었다. 배지의 pH가 4.0 혹은 7.0일때는 세포의 건물중이 현저히 감소하였고 배의 발생도 억제되었다. 그러나 pH 6에서 건물중이 최대에 달하였고 배의 발생도 양호하였다.

• 식물조직배양학회지
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• 제24권6호
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• pp.361-367
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• 1997

담배(Nicotiana tabacum cv. BY4)의 이배체식물과 담배의 약으로부터 유도된 반수체식물의 잎절편을 사용하여 2.4-D를 단독톄처리하여 캘러스를 유도하였다. 캘러스증식은 이배체에서는 2.0 mg/L 2.4-D와 1.0mg/L 카이네틴의 조합처리구에서, 반수체의 경우에 있어서는 2.0mg/L 2.4-D와 0.1mg/L BAP 조합처리구에서 실시하였으며, 현탁배양도 동일조건으로 실시하였다. 현탁배양 세포괴를 배양 15-20일째가 되면 0.8% low melting agarose와 혼합하여 카드a뮴이 0-2,000 $\mu$M의 농도로 처리된 선발배지에 평판배양하여 카드뮴저항성 colony(cell clump)를 조사관찰하였다. 그 결과 본 실험에서는 이배체는 세포의 성장에 있어서 최하저해농도(MIC : Minimum Inhibition Concentration)는 Cd$^{++}$ 300 $\mu$M이었으며 반수체는 Cd$^{++}$ 200 $\mu$M이었다. 이때 반수체 및 이배체의 MIC에서 선발된 세포주를 카드뮴 500, 1,000, 2,000 $\mu$M 농도로 옳겨 경화시킨 후 카드뮴저항성을 선발하였고 그 특성을 조사하였다. 그 결과 카드뮴 저항성을 띤 반수체 및 이배체유래 캘러스의 경우에는 각각 카드a뮴의 농도가 증가할수록 생중량은 감소하였으나, 건조중량은 오히려 증가하였다. 그리고 각각 선발된 세포주내의 카드뮴의 함량을 AAS를 사용하여 측정하였던 바, 카드뮴의 농도가 높을수록 캘러스에 축적된 카드뮴의 양이 높게 나타났다.

• KSBB Journal
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• 제13권5호
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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.