Expression of Pisum sativum defensin 1 (Psd1) in shaking flasks and bioreactor cultivations of recombinant Pichia pastoris at different pHs

AUTOR(ES)

Larentis, A. L., Almeida, M. S., Cabral, K. M. S., Medeiros, L. N., Kurtenbach, E., Coelho, M. A. Z.

FONTE

Brazilian Journal of Chemical Engineering

DATA DE PUBLICAÇÃO

2004-06

RESUMO

The methylotrophic yeast Pichia pastoris was used to produce the recombinant Pisum sativum defensin (rPsd1), a small peptide from pea seeds that has a high level of antifungal activity. The plasmid rPsd1/pPIC9 was integrated into the yeast genome and methanol was used to induce expression and secretion of the recombinant Psd1, at 30º C in a fed-batch mode. The effects of different pH conditions and process scale-up were evaluated using a Monod-type model where dissolved oxygen was considered the limiting substrate. Parameter estimation showed that the process could be improved by expressing rPsd1 in a 1000 mL bioreactor at pH 4. Structural and functional analyses revealed that the recombinant Psd1 is very similar to the native one.

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