This Article Full Text (PDF) Other Versions of this Article: AEM.02759-06v1 73/13/4342    most recent Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Verwaal, R. Articles by van Ooyen, A. J.J. Search for Related Content PubMed PubMed Citation Articles by Verwaal, R. Articles by van Ooyen, A. J.J. Agricola Articles by Verwaal, R. Articles by van Ooyen, A. J.J.

 Previous Article  |  Next Article 

Appl. Environ. Microbiol. doi:10.1128/AEM.02759-06
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

High level production of beta-carotene in Saccharomyces cerevisiae by successive transformation with carotenogenic genes from Xanthophyllomyces dendrorhous

Fungal Genomics, Laboratory of Microbiology, Wageningen University. Dreijenlaan 2, 6703, HA, Wageningen, the Netherlands; Biosynthesis Group, Molecular Biosciences 213, J.W. Goethe Universitaet, Siesmayerstr. 70, P.O: Box 111932, D-60054, Frankfurt, Germany

^* To whom correspondence should be addressed. Email: Ab.vanOoyen{at}wur.nl.

	Abstract

To determine whether Saccharomyces cerevisiae could serve as a host for efficient carotenoid and especially -carotene production, carotenogenic genes from the carotenoid producing yeast Xanthophyllomyces dendrorhous were introduced and overexpressed in S. cerevisiae. Because overexpression of these genes from an episomal expression vector resulted in instable strains, the genes were integrated into genomic DNA to yield stable, carotenoid producing S. cerevisiae cells. Furthermore, carotenoid production levels were higher in strains containing integrated carotenogenic genes. Overexpression of crtYB (encoding a bi-functional phytoene synthase and lycopene cyclase) and crtI (phytoene desaturase) from X. dendrorhous was sufficient to enable carotenoid production. Carotenoid production levels were increased by additional overexpression of a homologous geranylgeranyl diphosphate (GGPP) synthase from S. cerevisiae, encoded by BTS1. Combined overexpression of crtE (heterologous GGPP synthase) from X. dendrorhous with crtYB and crtI and the introduction of an additional copy of a truncated 3-hydroxy-3-methylglutaryl-coenzyme A reductase (tHMG1) into carotenoid producing cells resulted in a successive increase in carotenoid production levels. The mentioned strains produced high levels of intermediates of the carotenogenic pathway and comparable low levels of the preferred end product -carotene, as determined by HPLC. We finally succeeded to construct a S. cerevisiae strain capable of producing high amounts of -carotene up to levels of 5.9 mg g^-1 dry weight^-1, which was accomplished by introduction of an additional copy of crtI and tHMG1 into carotenoid producing yeast cells. This transformant is promising for further development towards biotechnological production of -carotene by S. cerevisiae.

This article has been cited by other articles:

• Nevoigt, E. (2008). Progress in Metabolic Engineering of Saccharomyces cerevisiae. Microbiol. Mol. Biol. Rev. 72: 379-412 [Abstract] [Full Text]