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Title: Hybrid promoter engineering strategies in Yarrowia lipolytica: isoamyl alcohol production as a test study
Authors: Zhao, Yu
Liu, Shiqi
Lu, Zhihui
Zhao, Baixiang
Wang, Shuhui
Zhang, Cuiying
Xiao, Dongguang
Foo, Jee Loon 
Yu, Aiqun
Keywords: Hybrid promoter
Isoamyl alcohol
Metabolic engineering
Synthetic promoter
Y. lipolytica
Issue Date: 2-Jul-2021
Publisher: BioMed Central Ltd
Citation: Zhao, Yu, Liu, Shiqi, Lu, Zhihui, Zhao, Baixiang, Wang, Shuhui, Zhang, Cuiying, Xiao, Dongguang, Foo, Jee Loon, Yu, Aiqun (2021-07-02). Hybrid promoter engineering strategies in Yarrowia lipolytica: isoamyl alcohol production as a test study. Biotechnology for Biofuels 14 (1) : 149. ScholarBank@NUS Repository.
Rights: Attribution 4.0 International
Abstract: Background: In biological cells, promoters drive gene expression by specific binding of RNA polymerase. They determine the starting position, timing and level of gene expression. Therefore, rational fine-tuning of promoters to regulate the expression levels of target genes for optimizing biosynthetic pathways in metabolic engineering has recently become an active area of research. Results: In this study, we systematically detected and characterized the common promoter elements in the unconventional yeast Yarrowia lipolytica, and constructed an artificial hybrid promoter library that covers a wide range of promoter strength. The results indicate that the hybrid promoter strength can be fine-tuned by promoter elements, namely, upstream activation sequences (UAS), TATA box and core promoter. Notably, the UASs of Saccharomyces cerevisiae promoters were reported for the first time to be functionally transferred to Y. lipolytica. Subsequently, using the production of a versatile platform chemical isoamyl alcohol as a test study, the hybrid promoter library was applied to optimize the biosynthesis pathway expression in Y. lipolytica. By expressing the key pathway gene, ScARO10, with the promoter library, 1.1–30.3 folds increase in the isoamyl alcohol titer over that of the control strain Y. lipolytica Po1g KU70? was achieved. Interestingly, the highest titer increase was attained with a weak promoter PUAS1B4-EXPm to express ScARO10. These results suggest that our hybrid promoter library can be a powerful toolkit for identifying optimum promoters for expressing metabolic pathways in Y. lipolytica. Conclusion: We envision that this promoter engineering strategy and the rationally engineered promoters constructed in this study could also be extended to other non-model fungi for strain improvement. © 2021, The Author(s).
Source Title: Biotechnology for Biofuels
ISSN: 1754-6834
DOI: 10.1186/s13068-021-02002-z
Rights: Attribution 4.0 International
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