Biological parts for Kluyveromyces marxianus synthetic biology

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dc.contributor.authorRajkumar, Arun S.
dc.contributor.authorVarela, Javier A.
dc.contributor.authorJuergens, Hannes
dc.contributor.authorDaran, Jean-Marc G.
dc.contributor.authorMorrissey, John P.
dc.contributor.funderHorizon 2020en
dc.contributor.funderSeventh Framework Programmeen
dc.contributor.funderBE-Basic Foundationen
dc.contributor.funderMinistry of Economic Affairs, Agriculture and Innovation, Netherlandsen
dc.date.accessioned2019-10-30T06:40:02Z
dc.date.available2019-10-30T06:40:02Z
dc.date.issued2019-05-07
dc.description.abstractKluyveromyces marxianus is a non-conventional yeast whose physiology and metabolism lends itself to diverse biotechnological applications. While the wild-type yeast is already in use for producing fragrances and fermented products, the lack of standardised tools for its genetic and metabolic engineering prevent it from being used as a next-generation cell factory for bio-based chemicals. In this paper, we bring together and characterise a set of native K. marxianus parts for the expression of multiple genes for metabolic engineering and synthetic biology. All parts are cloned and stored according to the MoClo/Yeast Tool Kit standard for quick sharing and rapid construction. Using available genomic and transcriptomic data, we have selected promoters and terminators to fine-tune constitutive and inducible gene expression. The collection includes a number of known centromeres and autonomously replication sequences (ARS). We also provide a number of chromosomal integration sites selected for efficiency or visible phenotypes for rapid screening. Finally, we provide a single-plasmid CRISPR/Cas9 platform for genome engineering and facilitated gene targeting, and rationally create auxotrophic strains to expand the common range of selection markers available to K. marxianus. The curated and characterised tools we have provided in this kit will serve as a base to efficiently build next-generation cell factories from this alternative yeast. Plasmids containing all parts are available at Addgene for public distribution.en
dc.description.sponsorshipMinistry of Economic Affairs, Agriculture and Innovation (FES subsidy to BE-Basic Research and Development)en
dc.description.statusPeer revieweden
dc.description.versionPublished Versionen
dc.format.mimetypeapplication/pdfen
dc.identifier.articleid97en
dc.identifier.citationRajkumar, A.S., Varela, J.A., Juergens, H., Daran, J.M. and Morrissey, J.P., 2019. Biological parts for Kluyveromyces marxianus synthetic biology. Frontiers in bioengineering and biotechnology, 7, 97. (15 pp.). DOI:10.3389/fbioe.2019.97en
dc.identifier.doi10.3389/fbioe.2019.97en
dc.identifier.eissn2296-4185
dc.identifier.endpage15en
dc.identifier.journaltitleFrontiers in Bioengineering and Biotechnologyen
dc.identifier.startpage1en
dc.identifier.urihttps://hdl.handle.net/10468/8924
dc.identifier.volume7en
dc.language.isoenen
dc.publisherFrontiers Media S.A.en
dc.relation.projectinfo:eu-repo/grantAgreement/EC/H2020::RIA/720824/EU/Model-Based Construction And Optimisation Of Versatile Chassis Yeast Strains For Production Of Valuable Lipid And Aromatic Compounds/CHASSYen
dc.relation.projectinfo:eu-repo/grantAgreement/EC/FP7::SP3::PEOPLE/606795/EU/Yeast Cell Factories: Training Researchers to Apply Modern Post-Genomic Methods In Yeast Biotechnology/YEASTCELLen
dc.relation.urihttps://www.frontiersin.org/articles/10.3389/fbioe.2019.00097
dc.rights© 2019 Rajkumar, Varela, Juergens, Daran and Morrissey. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.en
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/en
dc.subjectKluyveromycesen
dc.subjectSynthetic biologyen
dc.subjectMetabolic engineeringen
dc.subjectGenome engineeringen
dc.subjectYeasten
dc.titleBiological parts for Kluyveromyces marxianus synthetic biologyen
dc.typeArticle (peer-reviewed)en
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