title: One-step integration of multiple genes into the oleaginous yeast Yarrowia lipolytica.
abstract: Yarrowia lipolytica is an unconventional yeast, and is generally recognized as safe (GRAS). It provides a versatile fermentation platform that is used commercially to produce many added-value products. Here we report a multiple fragment assembly method that allows one-step integration of an entire β-carotene biosynthesis pathway (~11 kb, consisting of four genes) via in vivo homologous recombination into the rDNA locus of the Y. lipolytica chromosome. The highest efficiency was 21%, and the highest production of β-carotene was 2.2 ± 0.3 mg per g dry cell weight. The total procedure was completed in less than one week, as compared to a previously reported sequential gene integration method that required n weeks for n genes. This time-saving method will facilitate synthetic biology, metabolic engineering and functional genomics studies of Y. lipolytica.
authors.affiliation: College of Bioengineering, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, China, slgao@cibt.ac.cn.
authors.forename: Shuliang
authors.initials: S
authors.lastname: Gao
authors.forename: Linna
authors.initials: L
authors.lastname: Han
authors.forename: Li
authors.initials: L
authors.lastname: Zhu
authors.forename: Mei
authors.initials: M
authors.lastname: Ge
authors.forename: Sheng
authors.initials: S
authors.lastname: Yang
authors.forename: Yu
authors.initials: Y
authors.lastname: Jiang
authors.forename: Daijie
authors.initials: D
authors.lastname: Chen
mesh_terms: D053898:Biosynthetic Pathways / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics; D004275:DNA, Ribosomal / Q000235:genetics; D005827:Genetics, Microbial / Q000379:methods*; D059765:Homologous Recombination; D060847:Metabolic Engineering / Q000379:methods*; D008967:Molecular Biology / Q000379:methods*; D013997:Time Factors; D025062:Yarrowia / Q000235:genetics*; D019207:beta Carotene / Q000096:biosynthesis
chemical_list: D004271:DNA, Fungal; D004275:DNA, Ribosomal; D019207:beta Carotene