Dissolved oxygen (DO)-stat fed-batch culture, which allows a high cell density culture of microorganisms under constant DO conditions, was applied to anti-CRP single-chain variable fragment (scFv) production using recombinant Escherichia coli. The DO-stat fed-batch culture was successfully performed under various DO conditions for more than 50 h, resulting in increased scFv production from 0.5 to 0.8 g/L by flask and batch cultures to 2.8–3.0 g/L by the fed-batch culture under the conditions of 5–40% of DO saturation. The formation of inclusion bodies was effectively depressed during DO-stat fed-batch operation; consequently, the solubility of anti-CRP scFv was significantly improved from 36–43% by the flask and batch cultures to 96–98% by the DO-stat fed-batch culture under a wide range of DO conditions. From the kinetic analysis of fed-batch experiments, it was also found that the successful folding of anti-CRP scFv in the cytoplasm occurred when metabolic rates, such as the specific growth rate and specific glucose consumption rate, were relatively low. These results show that the fed-batch culture operated by the DO-stat feeding strategy was effective for the enhanced production of anti-CRP scFv with high solubility.

溶解氧 (DO)-stat 分批补料培养，允许在恒定 DO 条件下进行高细胞密度的微生物培养，用于使用重组大肠杆菌生产抗 CRP 单链可变片段 (scFv). DO-stat 分批补料培养在各种 DO 条件下成功进行了 50 小时以上，导致 scFv 产量从 0.5 到 0.8 g/L 的烧瓶和分批培养增加到 2.8–3.0 g/L 的补料分批DO 饱和度为 5-40% 的条件下培养。DO-stat补料分批操作有效抑制了包涵体的形成；因此，在各种溶解氧条件下，抗 CRP scFv 的溶解度从烧瓶和分批培养的 36-43% 显着提高到 DO-stat 补料分批培养的 96-98%。从分批补料实验的动力学分析还发现，当代谢率，例如比生长率和比葡萄糖消耗率相对较低时，抗CRP scFv在细胞质中的成功折叠发生。