• 식물조직배양학회지
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• 제24권5호
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• pp.279-284
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• 1997

Hyoscyamus muticus의 phytoalexin으로 알려진 solavetivone, lubimin, rishitin 생합성에 결정적 역할을 담당하는 isoprenoid pathway의 첫 분지효소인 vetispiradiene synthase의 유도특성과 solavetivone, lubimin phytoalexin 생합성과의 관련성을 구명하기 위하여 H. muticus 현탁세포배양 및 모상근배양에 Rhizoctonia solani 추출액을 elicitor로 처리하였다. Elicitor을 처리하지 않은 현탁세포 및 모상근배양에서는 효소활성, 효소단백질 및 solavetivone, lubimin등이 전혀 검출되지 않았으나, elicitor을 처리한 세포에서는 효소활성이 급속히 유도되어 처리 12시간 후에 최고의 활성을 보였으며 그 후 점점 감소하였다. 효소활성의 정도는 면역반응을 이용하여 측정한 효소 단백질의 함량과 밀접한 상관관계를 보였으며, phytoalexin 총 함량은 모상근배양에서 2배이상 높았다. 특히 solavetivone 및 lubimin 생합성은 현탁 및 모상근배양에서 서로 다른 특이성을 보였는데, 현탁배양에서는 lubimin을, 모상근배양에서는 solavetivone을 선택적으로 다량 생산하였다.

• Journal of Plant Biotechnology
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• 제3권2호
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• pp.107-112
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• 2001

The kinetics of Ti-transformed Salvia miltiorrhiza cell cultures was studied in 250-$m\ell$ shake flasks by using B5 medium with addition of 30 gfL of sucrose. In the cell cultures, the maximum cell mass obtained was 11.5 g DW/L on day 15. The highest amount of phenolic compounds - rosmarinic acid (RA) and lithospermic acid B (LAB) reached 871.3 mg/L (day 15) and 121.3 mg/L (day 13), respectively. The total tanshinone production, i.e., intracellular plus extracellular cryptotanshinone, tanshinone 1, and tanshinone IIA, was 5.3 mg/L on day 13. For the cultivations in 2.4-L stirred bioreactors, the residual sugar level and medium conductivity were a little higher in a small turbine impeller reactor ($T_s$) than those in a large turbine impeller reactor ($T_L$), while a higher cell density was obtained in the $T_L$. For the production of tanshinones and phenolics, better results were obtained in the $T_L$ than in the $T_s$. In the $T_L$, similar or even a little higher production titers of tanshinones and phenolic compounds were achieved compared to those in the flasks. The results suggest that the shake flask results could be successfully scaled up to the $T_L$ reactor. Such a large impeller reactor like $T_L$ may be better than a small impeller one for the large-scale production of the valuable metabolites by the suspension cultures of Ti transformed S.miltiorrhiza cells. This is considered due to the beneficial culture environment in the $T_L$, such as low shear rates as estimated theoretically.

• BMB Reports
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• 제32권5호
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• pp.451-455
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• 1999

The concentrations of L-ascorbic acid (AsA, ascorbate, vitamin C) and its biosynthetic and metabolically-related enzymes such as L-galactono-1,4-lactone dehydrogenase (GLDase), ascorbate peroxidase (APX), and ascorbate oxidase (ASO) were investigated in suspension cultures of Scutellaria baicalensis. Cells growing from 4 days after subculture (DAS) to 9 DAS and from 16 DAS to 19 DAS showed a diauxic growth, and then growth rapidly decreased with further culturing. The AsA content slowly increased to 19 DAS, reached a maximum at 21 DAS (ca $120\;{\mu}g/g$ dry cell wt), and then rapidly decreased with further culturing. GLDase and ASO activity were well correlated with the cell growth curve, showing a maximum at 19 DAS, whereas APX activity showed a good correlation with the changes in AsA content, showing a maximum at 21 DAS. The total ascorbate contents (reduced form, AsA, and oxidized form, dehydroascorbate, DHA) were markedly enhanced at 10 DAS when L-galactose and L-galactono-1,4-lactone (25 mM) were added to SH medium supplemented with 20 g/l sucrose at 9 DAS, by 5.5 and 6.8 times, respectively. DHA composed more than 90% of the total ascorbate contents in suspension cultures of S. baicalensis, even though the ratio of reduced to oxidized form slightly varied with cell growth stage. The results indicate that L-galactose and L-galactono-1,4-lactone are effective precursors of AsA in cell cultures of S. baicalensis, and that in vitro cultured cells provide suitable biomaterials for the study of biosynthesis and metabolism of AsA.

• 한국식물생명공학회:학술대회논문집
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• 한국식물생명공학회 2002년도 춘계학술대회
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• pp.159-159
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• 2002

• 한국식물생명공학회:학술대회논문집
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• 한국식물생명공학회 2002년도 춘계학술대회
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• pp.160-160
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• 2002

• Biotechnology and Bioprocess Engineering:BBE
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• 제6권1호
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• pp.51-55
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• 2001

This study was conducted to establish a plant cell culture system for the production of medically important secondary metabolites from Xanthium strumarium. The effects of plant growth regulators including NAA, 2,4-D, kinetin, and ABA were examined in terms of callus induction, maintenance of callus and suspension cultures. It was shown that callus was induced upon treatment with NAA while embryo was induced after treatment with 2,4-D. Callus formation was further improved by treatment with ABA and NAA. The level of callusing increased by 17-29% for the seed case, cotyledon, leaf, and hypocotyl and by 96% in the case of the root. Suspension cell lines were established using calli produced from cotyledon, hypocotyl and root and cultured at 25$\^{C}$ under light conditions. The cells grew up to 15g/L with NAA 2ppm, BA 2ppm, and ABA 1ppm treatment. Supernatants of suspension cultures of cell lines derived from coyledon and hypocotyl produced some distinctive secondary metabolites, one of which was identified as 8-epi-tomentosin, which belongs to the xanthanolides. The amounts of 8-epi-tomentosin produced by the cotyledon- and hypocotylderived cell lines were 13.4mg/L and 11.0mg/L, respectively.

• 한국식물생명공학회:학술대회논문집
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• 한국식물생명공학회 2003년도 식물바이오벤처 페스티발
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• pp.189-189
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• 2003

• KSBB Journal
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• 제15권6호
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• pp.560-565
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• 2000

13-Deacetyl-taxchinin I, a taxoid having a rearranged 11(15\longrightarrow1)-abeo-taxane skeleton, has been isolated and identified from plant cell cultures of Taxus chinensis. The compound has not previously been encountered in nature. Its structure was elucidated by 1-and 2D NMR techniques including H-H COSY, HMQC, and HMBC experiments. This taxoid also provides information for better understanding of structure-activity relationships and biosynthesis, as well as improving the quality control of paclitaxel production.

• Journal of Microbiology and Biotechnology
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• 제1권1호
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• pp.79-83
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• 1991

The effects of phosphate concentration in the medium, feeding of biosynthetic precursor, and the addition of exogenous berberine on cell growth and berberine production were studied in cell suspension cultures of Thalictrum rugosum. The depletion of phosphate in the medium enhanced the specific productivity up to twofold with significant release of berberine into the medium. Extracellular berberine was 19% of the total in the culture without phosphate while it was 2-5% of total berberine in the culture with even low amounts of phosphate. Precursor feeding was not effective in enhancing alkaloid formation. Initial presence of exogenous berberine did not have much effect on cell growth and alkaloid production. It was found that the cells have the capacity to take up large quantities of berberine. When $500{\;}mg{\cdot}l^{-1}$ of berberine was added exogenously at the beginning, 81% of total berberine was found in the cells.

• Journal of Plant Biotechnology
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• 제30권3호
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• pp.293-299
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• 2003

The effects of low dosage ${\gamma}$-radiation on the cell growth and the formation of shikonin derivatives were investigated in callus cultures of Lithospermum erythrorhizon under different medium and light conditions. Gamma radiation significantly affected the cell growed and formation of shikonin derivatives, depending on the culture conditions. In the cell cultures grown on M-9 medium, 2Gy and 16Gy of ${\gamma}$-radiation increased the calli growth and the formation of shikonin derivatives, respectively under 16hr day light condition. When calli were cultured for 60 days in the dark after irradiation of ${\gamma}$-radiation, cell growth was increased at low dosage of 1Gy and 2Gy in LS medium containing BA 2mg/L and IAA 0.2mg/L. Interestingly, calli grown in M-9 medium by 2Gy irradiation for 60 days significantly stimulated the formation of shikonin derivatives(13.21mg/g cell fresh wt), which was approximately 6 times higher than untreated cells.