雅罗维亚
发起人
生物
异源的
报告基因
酵母
基因
遗传学
基因表达
作者
Ioannis K. Georgiadis,Christina Tsiligkaki,Victoria Patavou,Maria Orfanidou,Antiopi Tsoureki,Aggeliki Andreadelli,Eleni Theodosiou,Antonios M. Makris
出处
期刊:Microorganisms
[MDPI AG]
日期:2023-04-28
卷期号:11 (5): 1152-1152
被引量:2
标识
DOI:10.3390/microorganisms11051152
摘要
Yarrowia lipolytica is a non-pathogenic aerobic yeast with numerous industrial biotechnology applications. The organism grows in a wide variety of media, industrial byproducts, and wastes. A need exists for molecular tools to improve heterologous protein expression and pathway reconstitution. In an effort to identify strong native promoters in glycerol-based media, six highly expressed genes were mined from public data, analyzed, and validated. The promoters from the three most highly expressed (H3, ACBP, and TMAL) were cloned upstream of the reporter mCherry in episomal and integrative vectors. Fluorescence was quantified by flow cytometry and promoter strength was benchmarked with known strong promoters (pFBA1in, pEXP1, and pTEF1in) in cells growing in glucose, glycerol, and synthetic glycerol media. The results show that pH3 > pTMAL > pACBP are very strong promoters, with pH3 exceeding all other tested promoters. Hybrid promoters were also constructed, linking the Upstream Activating Sequence 1B (UAS1B8) with H3(260) or TMAL(250) minimal promoters, and compared to the UAS1B8-TEF1(136) promoter. The new hybrid promoters exhibited far superior strength. The novel promoters were utilized to overexpress the lipase LIP2, achieving very high secretion levels. In conclusion, our research identified and characterized several strong Y. lipolytica promoters that expand the capacity to engineer Yarrowia strains and valorize industrial byproducts.
科研通智能强力驱动
Strongly Powered by AbleSci AI