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

开发了一种利用原生质体生产有用材料的高速系统。在微生物和植物原生质体中，一些产品被自由释放到肉汤中，从而产生了提高整体生产率和促进下游加工的双重后果。此外，由于许多原本不能被细胞接触到的高分子物质(由于细胞壁的存在)可以直接进入原生质体的细胞膜，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。研究发现，在液体静态培养条件下，酵母菌原生质体在发酵液中扩散细胞壁成分，在保持原生质体状态的同时，可以产生发酵液中的细胞壁成分。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。具有人工细胞壁的红豆杉原生质体高效地产生类鼻虫，并将化合物释放到培养基中。此外，适量供应茉莉酸甲酯等有利于提高红豆杉原生质体中紫杉类化合物的产量。这一新系统在生产有用的代谢物方面具有巨大的潜力。

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.