Accelerated prediction of recombinant protein production in Saccharomyces cerevisiae by using rapid monitoring techniques.
The use of a stopped-flow 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 promoter, and the monitoring facility is used to record the formation of the cutinase and cell density with optical density measurements. A range of induction conditions was studied with a view to using the monitoring to predict 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 modelling relating cutinase formation and cell production to predict likely final specific activities of the product. The utility of such monitoring and prediction is discussed with regards to improved process confidence and definition during fermentation production.
Brown
R
The Advanced Centre for Biochemical Engineering, University College London, Torrington Place, WC1E 7JE, London, UK
O'Kennedy
RD
Helwigh
J
Madden
E
Hoare
M