Direct product sequestration of a recombinant cutinase from batch fermentations of Saccharomyces cerevisiae.
The recovery of cutinase of Fusarium solani pisi produced by the yeast Saccharomyces cerevisiae was studied in a fluidised bed adsorption system directly integrated with a productive fermenter (so-called direct product sequestration; DPS). The relative efficiency of this system was compared with the one of a conventional purification process by discrete sequences of fermentation, broth clarification, ultrafiltration and fixed bed anion exchange chromatography. By direct product sequestration of the extracellular heterologous cutinase it was possible, through only one unit operation: (i) to perform broth clarification, (ii) to obtain a high cutinase concentration factor, and (iii) to recover cutinase with a specific activity that equalled that obtained with the conventional purification process. It was also possible (iv) to substantially reduce the total process time, (v) to improve the overall yield, and (vi) to increase cutinase productivity. Furthermore, the procedure outlined is suitable for large scale bioprocess exploitation.
Calado
C R
CR
Centro de Engenharia Biológica e Química, Instituto Superior Técnico, Lisboa, Portugal.
Hamilton
G E
GE
Cabral
J M
JM
Fonseca
L P
LP
Lyddiatt
A
A
D002265:Carboxylic Ester Hydrolases / Q000302:isolation & purification*; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D005285:Fermentation; D005670:Fusarium / Q000201:enzymology; D011994:Recombinant Proteins / Q000302:isolation & purification; D012441:Saccharomyces cerevisiae / Q000378:metabolism*
D011994:Recombinant Proteins; D002265:Carboxylic Ester Hydrolases; C019952:cutinase