Deciphering how LIP2 and POX2 promoters can optimally regulate recombinant protein production in the yeast Yarrowia lipolytica.

BACKGROUND

In recent years, species of non-conventional yeasts Model Yarrowia lipolytica has received much attention because it is a useful plant cells to produce recombinant proteins. In this species, the expression vector involving LIP2 and POX2 promoter has been developed and successfully used for the production of proteins on the same results with or even higher than other cell factories, such as Pichia pastoris. General Recombinant Proteins

 However, the production process involving the promoter can be difficult to manage, especially if done on a large scale in fed-batch bioreactor, because they require a hydrophobic inducers, such as oleic acid or methyl oleate. Thus, the challenge has been to reduce the burden on the hydrophobic substrate while promoting the production of recombinant proteins. One possible solution is to replace most of the inducer with co-substrates that can serve as an alternative energy source.

 However, applying such an approach would require a detailed knowledge of how the regulations impact a carbon source promoter, which surprisingly still less to LIP2 and POX2 promoter. The purpose of this study is to characterize the regulation thus better promoter and cell metabolism in Y. lipolytica cultures grown in media which are equipped with different carbon sources.

RESULTS

pPOX2 induction can be detected when glucose or glycerol is used as the sole carbon source, which means that the carbon source could not prevent the induction promoter. In addition, when a mixture of glucose and oleic acid are used in complex media, pPOX2 induction level lower that of pLIP2. 

In contrast, induction pLIP2 not present when glucose was present in the culture medium, which means that cell growth can occur without the recombinant gene expression. When a 40/60 mixture of glucose and oleic acid (w / w) is used, a tenfold increase in promoter induction, compared to when oleic-acid-only media observed. It was also obvious that the individual cells that adapt metabolically to use glucose and oleic acid. 

Indeed, there is no distinct subpopulations that are specific to glucose compared to oleic acid was observed; such an outcome would cause a producer and non-producer phenotype. In a medium containing glucose and oleic acid, the cells tend to metabolize oleic acid directly instead of storing it in the body lipids.

CONCLUSION

The study found that pLIP2 is compared Human Recombinant Proteins with pPOX2 promoter to drive expression of genes for the production of recombinant proteins by Y. lipolytica is used as a cell factory.