Expression of Pisum sativum defensin 1 (Psd1) in shaking flasks and bioreactor cultivations of recombinant Pichia pastoris at different pHs
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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| Main Authors: | , , , , , |
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| Format: | Digital revista |
| Language: | English |
| Published: |
Brazilian Society of Chemical Engineering
2004
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| Online Access: | http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0104-66322004000200004 |
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