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首页> 外文期刊>Applied Microbiology and Biotechnology >Biotransformation of tryptamine and secologanin into plant terpenoid indole alkaloids by transgenic yeast
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Biotransformation of tryptamine and secologanin into plant terpenoid indole alkaloids by transgenic yeast

机译：转基因酵母将色胺和secologanin生物转化为植物萜类吲哚生物碱

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摘要

A transgenic Saccharomyces cerevisiae was constructed containing the cDNAs coding for strictosidine synthase (STR) and strictosidine β-glucosidase (SGD) from the medicinal plant Catharanthus roseus. Both enzymes are involved in the biosynthesis of terpenoid indole alkaloids. The yeast culture was found to express high levels of both enzymes. STR activity was found both inside the cells (13.2 nkatal/g fresh weight) and in the medium (up to 25 nkatal/l medium), whereas SGD activity was present only inside the yeast cells (2.5 mkatal/g fresh weight). Upon feeding of tryptamine and secologanin, this transgenic yeast culture produced high levels of strictosidine in the medium; levels up to 2 g/l were measured. Inside the yeast cells strictosidine was also detected, although in much lower amounts (0.2 mg/g cells). This was due to the low permeability of the cells towards the substrates, secologanin and tryptamine. However, the strictosidine present in the medium was completely hydrolyzed to cathenamine, after permeabilizing the yeast cells. Furthermore, transgenic S. cerevisiae was able to grow on an extract of Symphoricarpus albus berries serving as a source for secologanin and carbohydrates. Under these conditions, the addition of tryptamine was sufficient for the transgenic yeast culture to produce indole alkaloids. Our results show that transgenic yeast cultures are an interesting alternative for the production of plant alkaloids.
机译：构建了转基因酿酒酵母，其含有来自药用植物长春花的编码严格糖苷合酶（STR）和严格糖苷β-葡萄糖苷酶（SGD）的cDNA。两种酶都参与萜类吲哚生物碱的生物合成。发现酵母培养物表达高水平的两种酶。在细胞内部（13.2纳卡特/克鲜重）和培养基（最高25纳卡特/升培养基）中均发现STR活性，而SGD活性仅在酵母细胞内部（2.5毫卡特/克鲜重）存在。喂食色胺和secologanin后，这种转基因酵母培养物在培养基中产生了高水平的严格的尿苷。最高可测量2 g / l。在酵母细胞内部也检测到了严格的核苷，尽管其含量低得多（0.2 mg / g细胞）。这是由于细胞对底物，次葡聚糖和色胺的渗透性低。然而，在使酵母细胞通透后，存在于培养基中的丁三醇被完全水解为联苯胺。此外，转基因酿酒酵母能够在Symphophcarpus albus浆果的提取物上生长，该提取物可作为secologanin和碳水化合物的来源。在这些条件下，色胺的添加足以使转基因酵母培养物产生吲哚生物碱。我们的结果表明，转基因酵母培养物是生产植物生物碱的有趣替代方法。

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  • 来源
    《Applied Microbiology and Biotechnology》 |2001年第4期|420-424|共5页
  • 作者

    A. Geerlings; F. Redondo; A. Contin; J. Memelink; R. van der Heijden; R. Verpoorte;

  • 作者单位

    Division of Pharmacognosy Leiden/Amsterdam Center for Drug Research Leiden University Gorlaeus Laboratories P.O. Box 9502 2300 RA Leiden The Netherlands;

    Division of Pharmacognosy Leiden/Amsterdam Center for Drug Research Leiden University Gorlaeus Laboratories P.O. Box 9502 2300 RA Leiden The Netherlands;

    Division of Pharmacognosy Leiden/Amsterdam Center for Drug Research Leiden University Gorlaeus Laboratories P.O. Box 9502 2300 RA Leiden The Netherlands;

    Institute of Molecular Plant Sciences Leiden University Clusius Laboratory Wassenaarseweg 64 2333 AL Leiden The Netherlands;

    Division of Pharmacognosy Leiden/Amsterdam Center for Drug Research Leiden University Gorlaeus Laboratories P.O. Box 9502 2300 RA Leiden The Netherlands;

    Division of Pharmacognosy Leiden/Amsterdam Center for Drug Research Leiden University Gorlaeus Laboratories P.O. Box 9502 2300 RA Leiden The Netherlands;

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