Utilization of protoplast for producing and screening of useful metabolites and it's application.

本文介绍了一种利用原生质体快速生产有用材料的系统。在微生物和植物原生质体中，一些产物自由释放到肉汤中，具有提高总体生产率和促进下游加工的双重后果。此外，由于许多高分子量物质（由于细胞壁的存在）不能直接进入细胞膜，因此原生质体产生的代谢物的组成和浓度与细胞产生的代谢物不同。发现在液体静态培养物中培养酵母原生质体，以使培养液中的细胞壁组分扩散，结果在保持原生质体状态的同时，原生质体可以在培养液中产生细胞壁组分。通过优化培养条件，获得了大量具有抗肿瘤活性的细胞壁成分。人工细胞壁的红豆杉原生质体能有效地产生紫杉醇类化合物并将其释放到培养基中。此外，茉莉酸甲酯等物质的充足供应有利于红豆杉原生质体紫杉烷类化合物的合成。这种新的系统在有用的代谢产物生产中具有很大的潜力。

A high-speed system for production of useful materials using protoplasts was developed. In microbial and plant protoplasts, some products are released freely into the broth with the double consequences of increasing overall productivity and facilitating down stream processing. Furthermore, since many high molecular weight substances which are otherwise not accessible to the cells (due to the presence of cell wall) can directly access the cell membrane in protoplasts, the composition and concentrations of the metabolites produced by protoplasts vary from those produced by cells. It was found that the yeast protoplasts are cultured in a liquid static culture so as to diffuse cell wall components in the culture broth with the result that while maintaining the state of the protoplasts, protoplasts could produce the cell wall components in the culture broth. By optimizing the culture conditions, large amount of the cell wall components with antitumor activity was produced. Taxus protoplasts with artificial cell walls produce taxiods efficiently and released the compounds into the medium. Moreover, the adequate supply of methyl-jasmonate, etc. were useful to enhance the taxoids production in Taxus protoplasts. This novel system has great potential in useful metabolite production.