An integrated scale-down plant for optimal recombinant enzyme production by Pichia pastoris |
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Authors: | Loegering Kai Mueller Christian Voss Jan-Patrick Wagenfuehrer Claudia Zahn Daniel Bertelsen Hans-Peter Scheffler Ulrich Luttmann Reiner |
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Institution: | Research Center of Bioprocess Engineering and Analytical Techniques, HAW - Hamburg University of Applied Sciences, Hamburg, Germany. |
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Abstract: | 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. |
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Keywords: | Automated enzyme production Cross-flow microfiltration Integrated bioprocess Lipase B from Candida antarctica Scale-down clarification |
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