BioChem - DOS

An integrated scale-down plant for optimal recombinant enzyme production by Pichia pastoris

Biotechnology journal

Loegering, Kai; Mueller, Christian; Voss, Jan-Patrick; Wagenfuehrer, Claudia; Zahn, Daniel; Bertelsen, Hans-Peter; Scheffler, Ulrich; Luttmann, Reiner

<P>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.</P>

2011

Wiley (John WileySons)

1860-6768

1860-7314

6

4

pp.428-436

10.1002/biot.201000384

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