Revisiting the unique structure of autonomously replicating sequences in Yarrowia lipolytica and its role in pathway engineering
Abstract
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 formore »
- Authors:
-
[3]
- Iowa State Univ., Ames, IA (United States)
- Iowa State Univ., Ames, IA (United States); Univ. of Illinois at Urbana-Champaign, IL (United States). Beckman Inst. for Advanced Science and Technology
- Ames Lab., and Iowa State Univ., Ames, IA (United States)
- Publication Date:
- Research Org.:
- Center for Advanced Bioenergy and Bioproducts Innovation (CABBI), Urbana, IL (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Biological and Environmental Research (BER); National Science Foundation (NSF)
- OSTI Identifier:
- 1834452
- Grant/Contract Number:
- SC0018420; 1716837; 1749782
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Applied Microbiology and Biotechnology
- Additional Journal Information:
- Journal Volume: 105; Journal Issue: 14-15; Journal ID: ISSN 0175-7598
- Publisher:
- Springer
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; plasmid stability; Yarrowia lipolytica; autonomously replicating sequence; plasmid loss and intramolecular rearrangements; genetic manipulation tools
Citation Formats
Lopez, Carmen, Cao, Mingfeng, Yao, Zhanyi, and Shao, Zengyi. Revisiting the unique structure of autonomously replicating sequences in Yarrowia lipolytica and its role in pathway engineering. United States: N. p., 2021.
Web. doi:10.1007/s00253-021-11399-4.
Lopez, Carmen, Cao, Mingfeng, Yao, Zhanyi, & Shao, Zengyi. Revisiting the unique structure of autonomously replicating sequences in Yarrowia lipolytica and its role in pathway engineering. United States. https://doi.org/10.1007/s00253-021-11399-4
Lopez, Carmen, Cao, Mingfeng, Yao, Zhanyi, and Shao, Zengyi. Fri .
"Revisiting the unique structure of autonomously replicating sequences in Yarrowia lipolytica and its role in pathway engineering". United States. https://doi.org/10.1007/s00253-021-11399-4. https://www.osti.gov/servlets/purl/1834452.
@article{osti_1834452,
title = {Revisiting the unique structure of autonomously replicating sequences in Yarrowia lipolytica and its role in pathway engineering},
author = {Lopez, Carmen and Cao, Mingfeng and Yao, Zhanyi and Shao, Zengyi},
abstractNote = {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.},
doi = {10.1007/s00253-021-11399-4},
journal = {Applied Microbiology and Biotechnology},
number = 14-15,
volume = 105,
place = {United States},
year = {Fri Aug 06 00:00:00 EDT 2021},
month = {Fri Aug 06 00:00:00 EDT 2021}
}
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