Accelerated prediction of recombinant protein production in Saccharomyces cerevisiae by using rapid monitoring techniques

The use of a stopped-how analyser for the monitoring of the production of a secreted recombinant protein, a wild-type cutinase, from S. cerevisiae CEN .PK111-32D pUR7320 is described. Induction is through use of a galactose pr omoter, and the monitoring facility is used to record the formation of the cutinase and cell density with optical density measurements. A range of ind uction conditions was studied with a view to using the monitoring to predic t the likely level of cutinase formation. Results achieved within 4 to 5 h of induction were of sufficient quality to allow the use of simple modellin g relating cutinase formation and cell production to predict likely final s pecific activities of the product. The utility of such monitoring and predi ction is discussed with regards to improved process confidence and definiti on during fermentation production. (C) 2000 Elsevier Science Inc. All right s reserved.