Journal
BIOCHEMICAL ENGINEERING JOURNAL
Volume 46, Issue 3, Pages 271-277Publisher
ELSEVIER
DOI: 10.1016/j.bej.2009.05.018
Keywords
Lip2; Candida rugosa; Pichia pastoris; AOX1 promoter; Enzyme; On-line fed-batch; Stirred tanks; Protein engineering
Funding
- Spanish program on Chemical Science and Technology [CTQ2007-60347/PPQ]
- DURSI (Generalitat de Catalunya)
- [2005-SGR-00698]
Ask authors/readers for more resources
The LIP2 isoenzyme gene from Candida rugosa has been completely synthesised and functionally expressed under the AOX1 promoter control in Pichia pastoris. The on-line monitoring and control of methanol, the key inducer carbon source in fed-batch cultures, has enhanced the yield product/biomass 7.8-fold and the productivity 12.8-fold compared to the best batch cultivation with the Pichia system and, 10-fold compared to the fed-batch cultivation process using the native C. rugosa strain. Nevertheless, the high ionic strength of culture broth favoured aggregation of Lip2, leading to total loss of lipolytic activity. After cultivation, a diaultrafiltration process was implemented to diminish ionic strength, allowing for the recovery of lipolytic activity in the diaultrafiltrate. The developed bioprocess resulted into a reproducible product in terms of quality and productivity. (C) 2009 Elsevier B.V. All rights reserved.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available