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Effect of Tween-80 on Stability and Secretion of Hydrophobic Tagged-cutinases

C. R. C. Calado ; Faculty of Engineering, Catholic University of Portugal, Estrada Octávio Pato, 2635-631 Rio de Mouro, Portugal
M. Brandão ; Institute for Biotechnology and Bioengineering, Instituto Superior Técnico, Av Rovisco Pais, 1049-001 Lisboa, Portugal
J. Biscaia ; Institute for Biotechnology and Bioengineering, Instituto Superior Técnico, Av Rovisco Pais, 1049-001 Lisboa, Portugal
J. M. S. Cabral ; Institute for Biotechnology and Bioengineering, Instituto Superior Técnico, Av Rovisco Pais, 1049-001 Lisboa, Portugal
L. P. Fonseca ; Institute for Biotechnology and Bioengineering, Instituto Superior Técnico, Av Rovisco Pais, 1049-001 Lisboa, Portugal


Puni tekst: engleski pdf 1.406 Kb

str. 411-417

preuzimanja: 473

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Sažetak

To significantly enhance the downstream processing, two cutinase variants were constructed by genetic fusion small hydrophobic peptides (WP)2 and (WP)4, respectively. However, the fusion of these peptides impairs cutinase secretion by the host cell
Saccharomyces cerevisiae and increases cutinase inactivation in the culture broth due to cutinase aggregation, resulting in cutinase activities per biomass of 56 % of cutinase-(WP)2 and of 7 % of cutinase-(WP)4, in relation to cutinase without the hydrophobic tags. It was observed that the addition of non-ionic surfactant Tween-80 into the culture broth could minimise the cutinase inactivation. The addition of Tween-80 also results in the enhancement of cutinase secretion by the yeast cell, leading to 1.25 and 2.51 fold-higher extracellular cutinase-(WP)2 and cutinase-(WP)4, respectively, in relation to cultivations performed in the absence of surfactant. Therefore, the addition of Tween-80 on the culture broth partly minimises the effect of fusion of the hydrophobic tags on the
inactivation of the enzymatic activity and on the reduction of the protein secretion. By this way, the use of Tween-80 on the S. cerevisiae cultivation may contribute to the efficiency enhancement of the downstream processing of tagged cutinases.

Ključne riječi

Cutinase, tagged-cutinase, Tween-80, secretion

Hrčak ID:

45380

URI

https://hrcak.srce.hr/45380

Posjeta: 809 *