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

Paper ID Volume ID Publish Year Pages File Format Full-Text
3997 203 2009 7 PDF Available
Title
Recombinant Candida rugosa LIP2 expression in Pichia pastoris under the control of the AOX1 promoter
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.

Keywords
Lip2; Candida rugosa; Pichia pastoris; AOX1 promoter; Enzyme; On-line fed-batch; Stirred tanks; Protein engineering
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Recombinant Candida rugosa LIP2 expression in Pichia pastoris under the control of the AOX1 promoter
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Publisher
Database: Elsevier - ScienceDirect
Journal: Biochemical Engineering Journal - Volume 46, Issue 3, 1 November 2009, Pages 271–277
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us