Elsevier

Metabolic Engineering

Volume 72, July 2022, Pages 1-13
Metabolic Engineering

Modular, synthetic chromosomes as new tools for large scale engineering of metabolism

https://doi.org/10.1016/j.ymben.2021.12.013Get rights and content
Under a Creative Commons license
open access

Highlights

  • Successful two-step modular, in vivo assembly of specialized Neochromosomes from 44 DNA parts.

  • Successful de novo production of pelargonidin 3-O-glucoside from glucose carried by a single Neochromosome.

  • Neochromosomes enable to add new functionalities, but also to remodel native functions.

  • ‘Anthocyanin’ Neochromosomes are stably replicated and segregated, and maintained at one copy.

  • Circular neochromosomes, more efficient to assemble than linear ones, are promising tools for metabolic engineering.

Abstract

The construction of powerful cell factories requires intensive genetic engineering for the addition of new functionalities and the remodeling of native pathways and processes. The present study demonstrates the feasibility of extensive genome reprogramming using modular, specialized de novo-assembled neochromosomes in yeast. The in vivo assembly of linear and circular neochromosomes, carrying 20 native and 21 heterologous genes, enabled the first de novo production in a microbial cell factory of anthocyanins, plant compounds with a broad range of pharmacological properties. Turned into exclusive expression platforms for heterologous and essential metabolic routes, the neochromosomes mimic native chromosomes regarding mitotic and genetic stability, copy number, harmlessness for the host and editability by CRISPR/Cas9. This study paves the way for future microbial cell factories with modular genomes in which core metabolic networks, localized on satellite, specialized neochromosomes can be swapped for alternative configurations and serve as landing pads for the addition of functionalities.

Keywords

Saccharomyces cerevisiae
Designer chromosome
Genome engineering
Metabolic engineering
Anthocyanin

Cited by (0)

1

Eline D. Postma and Else-Jasmijn Hassing contributed equally to this publication and should be considered co-first authors.