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

Lookup NU author(s): Dr Ronan O'Kennedy

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Abstract

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.

Publication metadata

Author(s): Brown RA, O'Kennedy RD, Helwigh J, Madden T, Hoare M

Publication type: Article

Publication status: Published

Journal: Enzyme and Microbial Technology

Year: 2000

Volume: 26

Issue: 9-10

Pages: 801-807

ISSN (print): 0141-0229

Publisher: Elsevier Inc.

URL: http://dx.doi.org/10.1016/S0141-0229(00)00174-5

DOI: 10.1016/S0141-0229(00)00174-5

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