Unconventional oleaginous yeast Yarrowia lipolytica has gained widespread applications as a microbial cell factory for synthesizing various chemicals and natural products.The construction of efficient cell factories r...Unconventional oleaginous yeast Yarrowia lipolytica has gained widespread applications as a microbial cell factory for synthesizing various chemicals and natural products.The construction of efficient cell factories re-quires intricate metabolic engineering.However,multi-gene expression in Y.lipolytica is labor-intensive.To facilitate multi-gene expression,we developed the polycistronic expression tool using 2A peptides.We first compared different 2A peptides in Y.lipolytica and identified two 2A peptides with high cleavage efficiency:P2A and ERBV-1.The effect of 2A peptides on the expression level of upstream and downstream genes was then determined.Ultimately,we applied the identified 2A peptides to express four genes in canthaxanthin biosyn-thetic pathway within one expression cassette for canthaxanthin production.This study enriches the multi-gene expression tools of Y.lipolytica,which will facilitate the cell factory construction of Y.lipolytica.展开更多
基金supported by the Key R&D Program of Shandong Province(2022SFGC0102)the Shandong Provincial Natural Science Foundation(ZR2022ZD24)the Taishan Scholars Program(tsqn202312061).
文摘Unconventional oleaginous yeast Yarrowia lipolytica has gained widespread applications as a microbial cell factory for synthesizing various chemicals and natural products.The construction of efficient cell factories re-quires intricate metabolic engineering.However,multi-gene expression in Y.lipolytica is labor-intensive.To facilitate multi-gene expression,we developed the polycistronic expression tool using 2A peptides.We first compared different 2A peptides in Y.lipolytica and identified two 2A peptides with high cleavage efficiency:P2A and ERBV-1.The effect of 2A peptides on the expression level of upstream and downstream genes was then determined.Ultimately,we applied the identified 2A peptides to express four genes in canthaxanthin biosyn-thetic pathway within one expression cassette for canthaxanthin production.This study enriches the multi-gene expression tools of Y.lipolytica,which will facilitate the cell factory construction of Y.lipolytica.
