Construction of a novel Pichia pastoris strain for production of xanthophylls

• Published in: AMB Express 2012-04, Vol.2 (1), p.24-24, Article 24
• Main Authors: Araya-Garay, José Miguel, Ageitos, José M, Vallejo, Juan A, Veiga-Crespo, Patricia, Sánchez-Pérez, Angeles, Villa, Tomás G
• Format: Article
• Language: English
• Subjects: Biomedical and Life Sciences; Biotechnology; Life Sciences; Microbial Genetics and Genomics; Microbiology; Original; Original Article
• ISSN: 2191-0855; 2191-0855
• DOI: 10.1186/2191-0855-2-24

In this study, we used the yeast carotenogenic producer Pichia pastoris Pp-EBIL strain, which has been metabolically engineered, by heterologously expressing β-carotene-pathway enzymes to produce β-carotene, as a vessel for recombinant astaxanthin expression. For this purpose, we designed new P. pastoris recombinant-strains harboring astaxanthin-encoding genes from carotenogenic microorganism, and thus capable of producing xanthophyllic compounds. We designed and constructed a plasmid (pGAPZA-WZ) containing both the β-carotene ketolase ( crtW ) and β-carotene hydroxylase ( crtZ ) genes from Agrobacterium aurantiacum , under the control of the GAP promoter and containing an AOX-1 terminator. The plasmid was then integrated into the P. pastoris Pp-EBIL strain genomic DNA, producing clone Pp-EBILWZ. The recombinant P. pastoris (Pp-EBILWZ) cells exhibited a strong reddish carotenoid coloration and were confirmed, by HPLC, to produce not only the previous described carotenoids lycopene and β-carotene, but also de novo synthesized astaxanthin.