Campus Units

Chemical and Biological Engineering, Microbiology, Ames Laboratory, NSF Engineering Research Center for Biorenewable Chemicals

Document Type

Article

Publication Version

Submitted Manuscript

Publication Date

12-2-2020

Journal or Book Title

Authorea

DOI

10.22541/au.160691063.38058320/v1

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. Our recent studies on the autonomously replicating sequences (ARSs) of nonconventional yeasts revealed that the ARSs fromY. 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 4.5-fold increase in gene expression and 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.

Comments

This is a pre-print of the article Lopez, Carmen, Mingfeng Cao, Zhanyi Yao, and Zengyi Shao. Investigating the role of noncoding regulatory DNA in plasmid development for Yarrowia lipolytica. Authorea (2020). DOI: 10.22541/au.160691063.38058320/v1. Posted with permission.

Copyright Owner

The Author(s)

Language

en

File Format

application/pdf

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