title: Construction of a novel Pichia pastoris strain for production of xanthophylls.
abstract: 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.
authors.affiliation: Department of Microbiology, University of Santiago de Compostela, Santiago de Compostela, Spain. tom.villa@gmail.com.
authors.forename: José Miguel
authors.initials: JM
authors.lastname: Araya-Garay
authors.forename: José M
authors.initials: JM
authors.lastname: Ageitos
authors.forename: Juan A
authors.initials: JA
authors.lastname: Vallejo
authors.forename: Patricia
authors.initials: P
authors.lastname: Veiga-Crespo
authors.forename: Angeles
authors.initials: A
authors.lastname: Sánchez-Pérez
authors.forename: Tomás G
authors.initials: TG
authors.lastname: Villa