One-step integration of multiple genes into the oleaginous yeast Yarrowia lipolytica

• DOI: 10.1007/s10529-014-1634-y
• Publication Date: 2014-09-12T05:40:48Z
• Authors: Shuliang Gao; Linna Han; Li Zhu; Mei Ge; Sheng Yang; Yu Jiang; Daijie Chen
• Keywords: Yarrowia; Metabolic Engineering; Synthetic Biology

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.