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Metabolic engineering of Pichia pastoris X-33 for lycopene production

Paper ID Volume ID Publish Year Pages File Format Full-Text
35060 45071 2009 8 PDF Available
Title
Metabolic engineering of Pichia pastoris X-33 for lycopene production
Abstract

As an alternative carotenoid producer, non-carotenogenic Pichia pastoris was chosen for a reddish carotenoid lycopene production because it can grow to high cell density without accumulation of ethanol and utilize various classes of organic materials such as methanol as carbon sources. Two synthetic lycopene-pathway plasmids, pGAPZB-EBI* and pGAPZB-EpBpI*p, were designed and constructed. The pGAPZB-EpBpI*p plasmid encoded three carotenogenic enzymes that were engineered to be targeted into peroxisomes of P. pastoris whereas the pGAPZB-EBI* plasmid encoded non-targeted enzymes. After both plasmids were transformed into P. pastoris, the lycopene-producing clone containing the pGAPZB-EpBpI*p plasmid, referred to as Ω, was selected and used for further optimization study. Of the carbon sources tested, glucose resulted in the highest level of lycopene production in complex and minimal media. Batch fermentation of the Ω clone resulted in the production of 4.6 mg-lycopene/g-DCW, with a concentration of 73.9 mg/l of lycopene in minimal medium. For the first time non-carotenogenic yeast P. pastoris was metabolically engineered by heterologously expressing lycopene-pathway enzymes and the lycopene concentration of 73.9 mg/l was obtained. This serves as a basis for the development of biological process for carotenoids using P. pastoris at a commercial production level.

Keywords
Lycopene; Pichia pastoris; Metabolic engineering; Carbon source; Carotenoids
First Page Preview
Metabolic engineering of Pichia pastoris X-33 for lycopene production
Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 44, Issue 10, October 2009, Pages 1095–1102
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering