Effect of Saccharomyces cerevisiae fermentation conditions on expanded bed adsorption of heterologous cutinase

✍ Scribed by Cecília R C Calado; Joaquim M S Cabral; Luis P Fonseca

• Publisher: Wiley (John Wiley & Sons)
• Year: 2002
• Tongue: English
• Weight: 118 KB
• Volume: 77
• Category: Article
• ISSN: 0268-2575
• DOI: 10.1002/jctb.695

✦ Synopsis

Abstract

The effect of culture media composition, and fermentation conditions and strategy on the growth and cutinase production of recombinant Saccharomyces cerevisiae and subsequent cutinase purification by expanded bed adsorption (EBA) was studied. The reduction in the amount of yeast extract used as nitrogen source from 20 g dm^−3^ to 10 g dm^−3^ in batch cultures led to a 29% decrease in the heterologous cutinase production, while the 5% cutinase dynamic adsorption capacity (__q__5%) on the cation Streamline SP XL was increased 6.7‐fold. By dilution of the whole fermentation broth, performed with the lowest yeast extract content, which reduces conductivity, the __q__5% was additionally increased by 1.9‐fold. After implementation of a fed‐batch strategy the cutinase concentration, cutinase yield on carbon source, cutinase yield on nitrogen source and productivity were increased by 10.8‐, 2.9‐, 5.3‐ and 6.4‐fold, respectively, in relation to the previously‐mentioned batch fermentation. However, the increased cutinase production was compromised by heterologous protein loss during the EBA recovery operation and the cutinase dynamic adsorption capacity and purification productivity decreased by 90% and 75%, respectively. Thus, target protein production by S cerevisiae fermentation and a downstream process with EBA cannot be considered as separate entities, where the understanding of the factors that affect the interactions among them are crucial towards optimization of the overall production process of heterologous proteins.

© 2002 Society of Chemical Industry