Construction of Platform Strains Producing Cosmeceutical Materials with Yarrowia Lipolytica
Yarrowia lipolytica which has been extensively used as the model oleaginous yeast for producing heterologous proteins has the capability to accumulate lipids (lipidbody, peroxisome etc) up to 50% of the DCW (dry cell weight). In addition, Y. lipolytica can be easily manipulated in gene deletion and insertion because it possesses a haploid genome and its genome sequence was completed and it is also known as GRAS (generally recognized as safe). Therefore, Y. lipolytica can be a high potential host strain in producing oleaginous (hydrophobic) and high value-added compounds used in cosmetic, medicine and nutraceutical materials which are hydrophobic and produced from plants by conventional extraction processes. Recently, these compounds have been produced in microorganism and it was well known that the DuPont company produced omega-3 using Y. lipolytica and industrialized it in 2010.
In this study, we used Y. lipolytica Po1g and constructed an ura3 auxotroph strain, Y. lipolytica (PolgDura3), in order to use URA3 blaster system which is a useful tool to carry out repeat genetic deletion and insertion on the genome. We also constructed a Y. lipolytica (PolgDku70Dura3) in which Ku70 involved in non-homologous recombination was deleted to increase recombination efficiency in this strain. Therefore, we confirm that Y. lipolytica (PolgDku70Dura3) showed higher homologous recombination efficiency (>5 folds) than Y. lipolytica (PolgDura3). In addition, we designed and constructed an expression vector pBJY_1 in which strong promoter, URA3 pop-out cassette and integrated target region were cloned for high expression of target genes and genome integration in Y. lipolytica. We synthesized target genes involved in biosynthesis of specific cosmetic and medicine materials and are cloning into the vector. We are sure that this strain and genetic tool are very useful to construct and develop various customized and industrial strains for producing various and valuable cosmeceuticals.