Revisiting the unique structure of autonomously replicating sequences in Yarrowia lipolytica and its role in pathway engineering

Lopez, Carmen; Cao, Mingfeng; Yao, Zhanyi; Shao, Zengyi

doi:10.1007/s00253-021-11399-4

Revisiting the unique structure of autonomously replicating sequences in Yarrowia lipolytica and its role in pathway engineering

Journal Article · Fri Aug 06 00:00:00 EDT 2021 · Applied Microbiology and Biotechnology

DOI:https://doi.org/10.1007/s00253-021-11399-4· OSTI ID:1834452

Lopez, Carmen ^[1]; Cao, Mingfeng ^[2]; Yao, Zhanyi ^[3]; ^[4]

Iowa State Univ., Ames, IA (United States); University of Illinois
Iowa State Univ., Ames, IA (United States); Univ. of Illinois at Urbana-Champaign, IL (United States). Beckman Inst. for Advanced Science and Technology
Iowa State Univ., Ames, IA (United States)
Ames Lab., and Iowa State Univ., Ames, IA (United States)

Production of industrially relevant compounds in microbial cell factories can employ either genomes or plasmids as an expression platform. Selection of plasmids as pathway carriers is advantageous for rapid demonstration but poses a challenge of stability. Yarrowia lipolytica has attracted great attention in the past decade for the biosynthesis of chemicals related to fatty acids at titers attractive to industry, and many genetic tools have been developed to explore its oleaginous potential. Here, our recent studies on the autonomously replicating sequences (ARSs) of nonconventional yeasts revealed that the ARSs from Y. lipolytica showcase a unique structure that includes a previously unannotated sequence (spacer) linking the origin of replication (ORI) and the centromeric (CEN) element and plays a critical role in modulating plasmid behavior. Maintaining a native 645-bp spacer yielded a 2.2-fold increase in gene expression and 1.7-fold higher plasmid stability compared to a more universally employed minimized ARS. Testing the modularity of the ARS sub-elements indicated that plasmid stability exhibits a pronounced cargo dependency. Instability caused both plasmid loss and intramolecular rearrangements. Altogether, our work clarifies the appropriate application of various ARSs for the scientific community and sheds light on a previously unexplored DNA element as a potential target for engineering Y. lipolytica.

View Accepted Manuscript (DOE)

Research Organization:: Center for Advanced Bioenergy and Bioproducts Innovation (CABBI), Urbana, IL (United States)

Sponsoring Organization:: National Science Foundation (NSF); USDOE Office of Science (SC), Biological and Environmental Research (BER)

Grant/Contract Number:: SC0018420

OSTI ID:: 1834452

Journal Information:: Applied Microbiology and Biotechnology, Journal Name: Applied Microbiology and Biotechnology Journal Issue: 14-15 Vol. 105; ISSN 0175-7598

Publisher:: SpringerCopyright Statement

Country of Publication:: United States

Language:: English

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Revisiting the unique structure of autonomously replicating sequences in Yarrowia lipolytica and its role in pathway engineering

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