Abstract

Metschnikowia pulcherrima is a non-conventional yeast with the potential to be used in bi-otechnological processes, especially involving low-cost feedstock exploitation. However, there are a lack of tools for researching it at a molecular level and for producing genetically modified strains. We tested the amenability to genetic modification of ten different strains, establishing a transformation protocol based on LiAc/PEG that allows us to introduce heterologous DNA. Non-homolo-gous integration was broadly successful and homologous recombination was successful in two strains. Chemical inhibition of non-homologous end joining recombination had a modest effect on the improvement of homologous recombination rates. Removal of selective markers via flippase recombinase was successful across integrated loci except for those targeted to the native URA3 lo-cus, suggesting that the genome sequence or structure alters the efficacy of this system.

Original languageEnglish
Article number290
Pages (from-to)1-11
Number of pages11
JournalMicroorganisms
Volume9
Issue number2
DOIs
Publication statusPublished - 31 Jan 2021

Bibliographical note

Funding Information:
Funding: This research was funded by the Industrial Biotechnology Catalyst (http://dx.doi.org/10.13039/501100006041, BBSRC and EPSRC to support the translation, development and commercialisation of innovative Industrial Biotechnology processes (EP/N013522/1) and via a URF studentship grant to M.M-B from the University of Bath.

Publisher Copyright:
© 2021 by the authors. Licensee MDPI, Basel, Switzerland.

Copyright:
Copyright 2021 Elsevier B.V., All rights reserved.

Keywords

  • Biotechnology
  • Homologous recombination
  • Nonconventional yeasts

ASJC Scopus subject areas

  • Microbiology
  • Virology
  • Microbiology (medical)

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