Recombinant Candida rugosa LIP2 expression in Pichia pastoris under the control of the AOX1 promoter

Pau Ferrer, Manuel Alarcón, Ramón Ramón, María Dolors Benaiges, Francisco Valero

Research output: Contribution to journalArticleResearchpeer-review

27 Citations (Scopus)

Abstract

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. © 2009 Elsevier B.V. All rights reserved.
Original languageEnglish
Pages (from-to)271-277
JournalBiochemical Engineering Journal
Volume46
DOIs
Publication statusPublished - 1 Nov 2009

Keywords

  • AOX1 promoter
  • Candida rugosa
  • Enzyme
  • Lip2
  • On-line fed-batch
  • Pichia pastoris
  • Protein engineering
  • Stirred tanks

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