Research Center of Bioprocess Engineering and Analytical Techniques, HAW - Hamburg University of Applied Sciences, Hamburg, Germany.
Biotechnol J. 2011 Apr;6(4):428-36. doi: 10.1002/biot.201000384.
An integrated bioprocess was created in a scale-down production plant by developing a two-stage enzyme production process with Pichia pastoris, containing a cell-breeding reactor and a production reactor in combination with a three-stage downstream process. To harvest the secreted enzymes, a disc separator and a cross-flow microfiltration clear the broth from the cells. Purification with hydrophobic interaction chromatography removes other proteins, concentrates the product, and prepares the enzyme solution for lyophilization. Fully automated and broad observable multi-stage parallel process courses have been developed using industrial process control systems and at-line measurements for enzyme concentration and enzyme activity. Optimal process conditions were found by application of Design of Experiments (DoE) for the production process.
采用毕赤酵母（Pichia pastoris）两阶段酶生产工艺，结合细胞培养罐和生产罐，以及三级下游处理工艺，在缩小规模的生产工厂中创建了一个集成式生物工艺。为了收获分泌的酶，采用碟式分离机和错流微滤从培养液中清除细胞。采用疏水相互作用色谱法进行纯化，可去除其他蛋白质，浓缩产物，并为冻干准备酶溶液。使用工业过程控制系统和在线测量酶浓度和酶活性，开发了全自动和宽观测多阶段平行过程课程。通过应用实验设计（DoE），找到了生产工艺的最佳工艺条件。
