• 식물조직배양학회지
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• 제27권4호
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• pp.269-282
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• 2000

Benzylisoquinoline alkaloids are a diverse group of natural products that include many pharmacologically active compounds produced in a limited number of plant families. Despite their complexity, intensive biochemical research has extended our knowledge of the chemistry and enzymology of many important benzylisoquinoline alkaloid pathways, such as those leading to the analgesic drugs morphine and codeine, and the antibiotics sanguinarine and berberine. The use of cultured plant cells as an experimental system has facilitated the identification and characterization of more than 30 benzylisoquinoline alkaloid biosynthetic enzymes, and the molecular cloning of the genes that encode at least 8 of these enzymes. The recent expansion of biochemical and molecular technologies has creat-ed unique opportunities to dissect the mechanisms involved in the regulation of benzylisoquinoline alkaloid biosynthesis in plants. Research has suggested that product accumulation is controlled by the developmental and inducible regulation of several benzylisoquinoline alkaloid biosynthetic genes, and by the subcellular compartmentation of biosynthetic enzymes and the intracellular localization and trafficking of pathway intermediates. In this paper, we review our current understanding of the biochemistry, cell biology, and molecular regulation of benzylisoquinoline alkaloid biosynthesis in plants. We also summarize our own research activities, especially those related to the establishment of protocols for the genetic transformation of benzylisoquinoline alkaloid-producing species, and the development of metabolic engineering strategies in these plants.

• Journal of Microbiology and Biotechnology
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• 제1권3호
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• pp.220-226
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• 1991

Large and rapid increases in benzophenanthridine alkaloid production occured in suspension cultures of Eschscholtzia californica cells treated with elicitors. Response to different biotic elicitors showed that elicitors prepared from yeast extract, Collectotrichum lindemuthianum and Verticillium dahliae induced alkaloid formation. Highest alkaloid accumulation was obtained with $60\;\mu\textrm{g}$ of yeast extract elicitor per gram of fresh cell weight. In time course performance after elicitor addition, more than 40 hours were required to obtain saturated alkaloid accumulation. Compounded silicone fluid, an ideal accumulation phase for two-phase culture of E. californica, accumulated a large amount of alkaloids produced in a specific manner. Elicitation in two-phase culture clearly increased net alkaloid production as well as their concentrations in the accumulation phase.

• KSBB Journal
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• 제11권4호
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• pp.411-419
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• 1996

본 연구에서는, 최근 들어 관심의 대상이 되어 의 품 산업에 응용의 수가 늘고 있는 cyclodextrin을 이용하여 세포밖으로 배출된 식물세포 2차 대사물질을 신속히 포획하여 생산수율을 증대시키고자 하였다. 실험결과 양귀비 현탁배양에서 a-, y-cyclodex trin보다는 ${\beta}$-cyclodextrin이 benzophenan-thridine alkaloid를 생산하는데 더 효과적업을 알았다. 이때 ${\beta}$-cyclodextrin의 optimum concentration이 존재하였는데, 그 농도는 현탁배양배지의 1.5% (w/v) 부 근에서였다. 고체 배지에서 ${\beta}$-cyclodextrin의 첨가는 sangumarme의 생산성을 control의 경우보다 약 4 40배(포함), 약 17배(살포) 정도 증가시켰다. ${\beta}$-cyclodextrin은 현탁배양 배지의 수용액상에서 세포 외로 배출된 alkaloid를 신속히 포획하여 inclusion complex를 형생함으로써 product의 배출을 촉진하여 결과적으로, product pattern을 변화시켜 줌으로 써 생산과 분리가 통시에 이루어질 수 있는 연속공정의 가능성을 확인시켜주었다. 또한 $\beta$cyclodex trin 1.5%(w/v)을 첨가시기를 달리하여 실험한 결과 배양초기에 넣어주는 것이 효과적이었으며, 이때 ${\beta}$-cyclodextrin은 세포성장에 미치는 영향이 없음을 확인하였다. ${\beta}$-cyclodextrin에 의한 benzophenan­t thridine alkaloid의 엄청난 생산 중대 효과는 1) In­c clusion complex 형성에 의한 안정성 증대뿐만 아니라 2) elicitor로서의 역할 등의 복합적 인 요인을 생 각해 볼 수 있는데 2)의 경우에 대한 실험을 세포와 ${\beta}$-cyclodextrin간의 직접적인 접촉을 시키지 않고 해본 결과 미세한 영향을 나타내었으므로 elicitation 의 경우를 완전히 배제한다고 할 수 없었다. 그러나 uv에 의한 inclusion complex의 확인이 1)의 경우를 확인하여 주었으므로 본 실험에서 ${\beta}$-cyclodex­t trin의 역할은 inclusion complex에 의한 생산성 증대라 할 수 있다.