Development of useful metabolites production system by protoplasts with artificial cell wall.

通过具有人造细胞壁的原生质体开发有用的代谢物生产系统。

• 批准号: 09555248
• 负责人: TANAKA Hideo
• 金额: $ 7.81万
• 依托单位: University of Tsukuba
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09555248/
• 关键词: artificial cell wall; plant protoplast; microbial protoplast; useful metabolites production; bioreactor; elicitor; alginate; inhibitor of cell wall synthesis; 細胞質合成阻害剤; 活性酸素; ガラクチュロン酸; 内在性エリシター; 微生物プロトプロスト; インドールアルカロイド; 固定化

Study on development of useful metabolites production system by microbial and plant protoplasts with artificial cell wall were carried out. The following results were obtained.(1) Discovery that alginate acts as an elicitor and can be used as an artificial cell wall.Various studies on the functions of alginate showed that alginate acts as an elicitor in promoting the production of many useful compounds by plant cells. The basic analysis of action mechanism of alginate was carried out, and it was postulated that there was a recognition site for alginate on cell membrane. It was found that in the protoplast which has no cell wall, the sensitivity of the alginate was high, and that the elicitor-effect remarkably increased. The elicitor-effect of alginate increased significantly when HィイD22ィエD2OィイD22ィエD2 and CaィイD12+ィエD1 were added simultaneously.(2) Development of simple method for preparation of microbial and plant protoplasts with artificial cell wall.After immobilization of cells in al … More ginate, the beads containing the immobilized cells were treated with cell wall digesting enzyme and protoplasts were formed inside the beads. This is a very simple and rapid method for producing both microbial and plant protoplasts with artificial cell walls.(3) The development of useful compound production system by the plant protoplasts with artificial cell wall.Catharanthus roseus protoplast were immobilized in alginate gel with the elicitor function as an artificial cell wall, and used for indole alkaloids production. In comparison with normal cells, the indole alkaloids production increased more than 30 times. Also system for cultivation of Wasabia japonica protoplast with artificial cell wall and with addition of HィイD22ィエD2OィイD22ィエD2 was developed. In this study, HィイD22ィエD2OィイD22ィエD2 and alginate acted synergistically and chitinase production increased by more than one order when comparison with normal cells.(4) Development of useful compound production system by the microbial protoplast with artificial cell wall.Protoplasts were prepared from yeast cells for production of invertase. Alginate gel with high mass transfer capacity was used as the artificial cell wall. When the protoplasts were cultivated in bubble column reactor, the invertase production increased much larger than that of the cells. Less

对利用人工细胞壁构建微生物和植物原生质体生产有用代谢物体系进行了研究。获得了以下结果。(1)海藻酸盐作为一种诱导子，可用作人工细胞壁的发现。对海藻酸盐功能的各种研究表明，海藻酸盐作为一种诱导子，可促进植物细胞产生许多有用的化合物。对海藻酸钠的作用机理进行了初步分析，认为海藻酸钠在细胞膜上存在识别位点。结果表明，在无细胞壁的原生质体中，藻酸盐的敏感性高，激发子的诱导效果显著增强。当同时加入H_2O_3、H_2O_4、H_4、H_2O_4、H_6、H_2O_4、H_2O (2)利用人工细胞壁制备微生物和植物原生质体的简便方法 ...更多信息 Ginate，用细胞壁消化酶处理含有固定化细胞的珠子，并在珠子内形成原生质体。这是一种非常简单和快速的方法，用于生产具有人工细胞壁的微生物和植物原生质体。(3)构建植物人工细胞壁原生质体生产有用化合物体系以长春花（Catharanthus roseus）原生质体为材料，利用诱导子作为人工细胞壁，将其固定在海藻酸钠凝胶中，用于吲哚生物碱的生产。与正常细胞相比，吲哚生物碱的产量增加了30倍以上。建立了用人工细胞壁和添加H_2O_3-D_22-D_2O-D_22-D_2的山葵原生质体培养体系。在本研究中，H_（22）O_（22 (4)利用具有人工细胞壁的微生物原生质体建立有用的复合物生产系统以生产转化酶的酵母细胞为原料制备原生质体。采用具有高传质能力的海藻酸凝胶作为人工细胞壁。原生质体在鼓泡塔反应器中培养时，转化酶产量的增加幅度远大于细胞本身。少

项目成果

H.Aoyagi and H.Tanaka et al.: "Development of simple methods for preparation of yeast and plant protoplasts immobilized in alginate gel beads"Biotechnology Techniques. 13(4). 253-258 (1999)

H.Aoyagi 和 H.Tanaka 等人：“开发固定在藻酸盐凝胶珠中的酵母和植物原生质体的简单制备方法”生物技术。

H.Aoyagi and H.Tanaka et al.: "Endogeneous elicitor-like effects of alginate on physiological activities of plant cells"Applied Microbiology and Biotechnology. 52(3). 429-436 (1999)

H.Aoyagi 和 H.Tanaka 等人：“藻酸盐对植物细胞生理活性的内源性诱导子样作用”应用微生物学和生物技术。

青柳 秀紀、田中 秀夫: "人工細胞壁を装着した酵母プロトプラストによるインベルターゼの生産" 化学工学会つくば化学技術懇話会 平成10年度研究交流会講演要旨. 34-35 (1999)

Hideki Aoyagi、Hideo Tanaka：“通过配备人工细胞壁的酵母原生质体生产转化酶”化学工程师学会筑波化学技术会议 1998 年研究交流会摘要 34-35（1999）。

H.Aoyagi and H.Tanaka et al.: "Alginate promotes production of various enzymes by Catharanthus roseus cells"Plant Cell Reports. 17(3). 243-247 (1998)

H.Aoyagi 和 H.Tanaka 等人：“藻酸盐促进长春花细胞产生各种酶”《植物细胞报告》。

H.Aoyagi and H.Tanaka et al.: "Synergistic effects of active oxygen species and alginate on chitinase production by Wasabia japonica cell and its application"Journal of Bioscience and Bioengineering. 89(2). 131-137 (2000)

H.Aoyagi 和 H.Tanaka 等：“活性氧和藻酸盐对山葵细胞产生几丁质酶的协同作用及其应用”生物科学与生物工程杂志。