Biotransformation with the plant cultured cells-Clarification and development of its enantioselective reactions

植物培养细胞的生物转化-其对映选择性反应的澄清和发展

• 批准号: 02804042
• 负责人: HIRATA Toshifumi
• 金额: $ 1.22万
• 依托单位: HIROSHIMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1990
• 资助国家: 日本
• 起止时间: 1990 至 1992
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02804042/
• 关键词: Plant cultured cells; Epoxidase; Reductase; Enantioselectivity; Substrate specificity; Stereospecificity; Biotransformation; Carbon-Carbon double bond; タバコ培養細胞; エノン系化合物; 炭素ー炭素二重結合

Biochemical potentiality of plant cultured cells to transform "foreign substrates" administered exogenously to the plant cell cultures is of considerable interest as a bioreactor. we have examined that the chiral introduction of hydrogen and oxygen atoms into the carbon-carbon double bond of terpene hydrocarbons with the enzyme preparation from the cultured cells of Nicotiana tabacum.1. Reduction of the C-C double bond of verbenone by a reductase from the cultured cells involves the syn addition of hydrogen atom from the re-re face of the double bond; the hydrogen atoms participating in the reduction at C-2 and C-3 of verbenone originates from the pro-4S hydrogen of NADPH and the medium, respectively.2. The cultured cells of N. tabacum have the ability to introduce enantioselectively the oxydenated of functions into the carbon-carbon double bond and its allylic position. Isolation and purification of a epoxydase from the cultured cells were performed. The epoxydase was found to discriminate the enantiotopic faces of the carbon-carbon double bonds and enantioselectively oxidize them from its re-re face.3. The bioconversion of carbone by the immobilized cells of N. tabacum entrapped in Ca alginate beads was investigated. Efficiency of the bioconversion by the immobilized cells was higher than that by the suspension cells. Only the reduction of the carbon-carbon double bond occurs in the medium of pH 7.5 and the reduction of the carbonyl group is prevented.

作为生物反应器，植物培养细胞转化外源给予植物细胞培养物的“外源底物”的生物化学潜力是相当感兴趣的。我们已经研究了用来自烟草培养细胞的酶制剂将氢和氧原子手性引入萜烯烃的碳-碳双键中。还原酶还原马鞭草烯酮的C-C双键时，双键表面的氢原子发生顺式加成，参与还原马鞭草烯酮C-2和C-3位的氢原子分别来自NADPH和培养基中的pro-4S氢.用N. tabacum具有将官能团的氧化物对映选择性地引入碳-碳双键及其烯丙基位置的能力。从培养的细胞中分离和纯化环氧酶。发现该环氧酶能够区分碳碳双键的对映异构面，并从其表面选择性地氧化碳碳双键.固定化N.海藻酸钙微球包埋烟草的研究。固定化细胞的转化效率高于悬浮细胞。在pH 7.5的介质中，仅发生碳-碳双键的还原，并且阻止了羰基的还原。

项目成果

S.IZUMI: "Synthesis of [10,10,10-^2H_3] Geranyl Diphosphate" J.Labelled Compd. Radiopharm.29. 591-597 (1991)

S.IZUMI：“[10,10,10-^2H_3]香叶基二磷酸的合成”J.Labeled Compd。

T. Hirata: "Enantioselective Introduction of Oxygen Functions into Alkenes. Oxydation of Limonene by the Cultured Suspension Cells of Nicotiana tabacum" Plant Tissue Culture Lett.10. (1993)

T. Hirata：“将氧功能对映选择性引入烯烃中。培养的烟草悬浮细胞对柠檬烯的氧化”植物组织培养快报 10。

T.Hirata, et al: "Volatile Monoterpenoid Constituents of the Plantlets of Mentha spicata Produced by Shoot Tip Culture" Phytochemistry. 29. 493-495 (1990)

T.Hirata 等人：“芽尖培养产生的留兰香植株的挥发性单萜成分”植物化学。

S. Izumi: "Synthesis of[10,10,10ー ^2H_3]Geranyl Diphosphate" J. Labelled Compd. Radiopharm.29. 591-597 (1991)

S. Izumi：“[10,10,10ー ^2H_3]香叶基二磷酸盐的合成”J. Labeled Compd.29 (1991)。

T.Suga,et al: "Biotransfor mation of Exogenous Substrates by Plant Cell Cultures" Phytochemistry. 29. 2393-2405 (1990)

T.Suga 等人：“植物细胞培养物对外源底物的生物转化”植物化学。