Increase in the astaxanthin synthase gene (crtS) dose by in vivo DNA fragment assembly in Xanthophyllomyces dendrorhous

BMC Biotechnology

Gabriela Contreras¹  Salvador Barahona¹  María Cecilia Rojas²  Marcelo Baeza¹  Víctor Cifuentes¹  Jennifer Alcaíno¹ 

[1] Departamento de Ciencias Ecológicas, Facultad de Ciencias, Universidad de Chile, Las Palmeras 3425, Casilla, Santiago 653, Chile

[2] Departamento de Química, Facultad de Ciencias, Universidad de Chile, Las Palmeras 3425, Casilla, Santiago 653, Chile

关键词: DNA assembler;    Astaxanthin synthase;    Xanthophyllomyces dendrorhous;   

Others  :  835303
DOI  :  10.1186/1472-6750-13-84

 received in 2013-07-12, accepted in 2013-10-04,  发布年份 2013

【 摘 要 】

Background

Xanthophyllomyces dendrorhous is a basidiomycetous yeast that is relevant to biotechnology, as it can synthesize the carotenoid astaxanthin. However, the astaxanthin levels produced by wild-type strains are low. Although different approaches for promoting increased astaxanthin production have been attempted, no commercially competitive results have been obtained thus far. A promising alternative to facilitate the production of carotenoids in this yeast involves the use of genetic modification. However, a major limitation is the few available molecular tools to manipulate X. dendrorhous.

Results

In this work, the DNA assembler methodology that was previously described in Saccharomyces cerevisiae was successfully applied to assemble DNA fragments in vivo and integrate these fragments into the genome of X. dendrorhous by homologous recombination in only one transformation event. Using this method, the gene encoding astaxanthin synthase (crtS) was overexpressed in X. dendrorhous and a higher level of astaxanthin was produced.

Conclusions

This methodology could be used to easily and rapidly overexpress individual genes or combinations of genes simultaneously in X. dendrorhous, eliminating numerous steps involved in conventional cloning methods.

【 授权许可】

   
2013 Contreras et al.; licensee BioMed Central Ltd.

【 图 表 】

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