Mosbach K, Birnbaum S, Hardy K, Davies J, Bülow L
Nature. 1983 Apr 7;302(5908):543-5. doi: 10.1038/302543a0.
There has been an increasing interest in the use of immobilized cells for the production of pharmaceuticals as well as for products such as high fructose syrup or ethanol. Some of these compounds are now produced on an industrial scale whereby the cells are used in a resting or growing state or in a nonviable form as natural carriers of the enzyme(s) involved in the synthesis. The advantages of immobilized cell technology should also apply to microorganisms modified by recombinant DNA techniques to produce a variety of eukaryotic proteins such as hormones. We describe here the properties of immobilized Bacillus subtilis cells carrying plasmids encoding rat proinsulin. Cell proliferation normally coupled to DNA replication is undesirable in immobilized cell systems as "clogging' of the system occurs due to cells growing outside the beads. Therefore, different ways were investigated to inhibit cell division while allowing continued protein synthesis. We found that the addition of certain antibiotics in the growth medium, such as novobiocin which inhibits DNA replication, fulfills these requirements, allowing proinsulin synthesis and excretion to take place over a period of several days.
人们对使用固定化细胞生产药物以及生产诸如高果糖糖浆或乙醇等产品的兴趣与日俱增。现在，其中一些化合物已实现工业化生产，在此过程中，细胞以静止、生长状态或无活性形式作为参与合成的酶的天然载体被使用。固定化细胞技术的优势也应适用于通过重组DNA技术改造的微生物，以生产多种真核蛋白，如激素。我们在此描述携带编码大鼠胰岛素原质粒的固定化枯草芽孢杆菌细胞的特性。在固定化细胞系统中，通常与DNA复制相关联的细胞增殖是不可取的，因为细胞在珠子外部生长会导致系统“堵塞”。因此，人们研究了不同的方法来抑制细胞分裂，同时允许蛋白质持续合成。我们发现，在生长培养基中添加某些抗生素，如抑制DNA复制的新生霉素，可满足这些要求，使胰岛素原在数天内得以合成和分泌。
