Engineering <named-content content-type="genus-species">Kluyveromyces marxianus</named-content> as a Robust Synthetic Biology Platform Host

ABSTRACT Throughout history, the yeast Saccharomyces cerevisiae has played a central role in human society due to its use in food production and more recently as a major industrial and model microorganism, because of the many genetic and genomic tools available to probe its biology. However, S. cere...

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Autores principales:	Paul Cernak, Raissa Estrela, Snigdha Poddar, Jeffrey M. Skerker, Ya-Fang Cheng, Annika K. Carlson, Berling Chen, Victoria M. Glynn, Monique Furlan, Owen W. Ryan, Marie K. Donnelly, Adam P. Arkin, John W. Taylor, Jamie H. D. Cate
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2018
Materias:	CRISPR-Cas9 Kluyveromyces marxianus lipogenesis mating renewable chemicals thermotolerant yeast Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/1ee3537f5483405a9dd2c47cbb421cca
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spelling	oai:doaj.org-article:1ee3537f5483405a9dd2c47cbb421cca2021-11-15T15:58:20ZEngineering <named-content content-type="genus-species">Kluyveromyces marxianus</named-content> as a Robust Synthetic Biology Platform Host10.1128/mBio.01410-182150-7511https://doaj.org/article/1ee3537f5483405a9dd2c47cbb421cca2018-11-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01410-18https://doaj.org/toc/2150-7511ABSTRACT Throughout history, the yeast Saccharomyces cerevisiae has played a central role in human society due to its use in food production and more recently as a major industrial and model microorganism, because of the many genetic and genomic tools available to probe its biology. However, S. cerevisiae has proven difficult to engineer to expand the carbon sources it can utilize, the products it can make, and the harsh conditions it can tolerate in industrial applications. Other yeasts that could solve many of these problems remain difficult to manipulate genetically. Here, we engineered the thermotolerant yeast Kluyveromyces marxianus to create a new synthetic biology platform. Using CRISPR-Cas9 (clustered regularly interspaced short palindromic repeats with Cas9)-mediated genome editing, we show that wild isolates of K. marxianus can be made heterothallic for sexual crossing. By breeding two of these mating-type engineered K. marxianus strains, we combined three complex traits—thermotolerance, lipid production, and facile transformation with exogenous DNA—into a single host. The ability to cross K. marxianus strains with relative ease, together with CRISPR-Cas9 genome editing, should enable engineering of K. marxianus isolates with promising lipid production at temperatures far exceeding those of other fungi under development for industrial applications. These results establish K. marxianus as a synthetic biology platform comparable to S. cerevisiae, with naturally more robust traits that hold potential for the industrial production of renewable chemicals. IMPORTANCE The yeast Kluyveromyces marxianus grows at high temperatures and on a wide range of carbon sources, making it a promising host for industrial biotechnology to produce renewable chemicals from plant biomass feedstocks. However, major genetic engineering limitations have kept this yeast from replacing the commonly used yeast Saccharomyces cerevisiae in industrial applications. Here, we describe genetic tools for genome editing and breeding K. marxianus strains, which we use to create a new thermotolerant strain with promising fatty acid production. These results open the door to using K. marxianus as a versatile synthetic biology platform organism for industrial applications.Paul CernakRaissa EstrelaSnigdha PoddarJeffrey M. SkerkerYa-Fang ChengAnnika K. CarlsonBerling ChenVictoria M. GlynnMonique FurlanOwen W. RyanMarie K. DonnellyAdam P. ArkinJohn W. TaylorJamie H. D. CateAmerican Society for MicrobiologyarticleCRISPR-Cas9Kluyveromyces marxianuslipogenesismatingrenewable chemicalsthermotolerant yeastMicrobiologyQR1-502ENmBio, Vol 9, Iss 5 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	CRISPR-Cas9 Kluyveromyces marxianus lipogenesis mating renewable chemicals thermotolerant yeast Microbiology QR1-502
spellingShingle	CRISPR-Cas9 Kluyveromyces marxianus lipogenesis mating renewable chemicals thermotolerant yeast Microbiology QR1-502 Paul Cernak Raissa Estrela Snigdha Poddar Jeffrey M. Skerker Ya-Fang Cheng Annika K. Carlson Berling Chen Victoria M. Glynn Monique Furlan Owen W. Ryan Marie K. Donnelly Adam P. Arkin John W. Taylor Jamie H. D. Cate Engineering <named-content content-type="genus-species">Kluyveromyces marxianus</named-content> as a Robust Synthetic Biology Platform Host
description	ABSTRACT Throughout history, the yeast Saccharomyces cerevisiae has played a central role in human society due to its use in food production and more recently as a major industrial and model microorganism, because of the many genetic and genomic tools available to probe its biology. However, S. cerevisiae has proven difficult to engineer to expand the carbon sources it can utilize, the products it can make, and the harsh conditions it can tolerate in industrial applications. Other yeasts that could solve many of these problems remain difficult to manipulate genetically. Here, we engineered the thermotolerant yeast Kluyveromyces marxianus to create a new synthetic biology platform. Using CRISPR-Cas9 (clustered regularly interspaced short palindromic repeats with Cas9)-mediated genome editing, we show that wild isolates of K. marxianus can be made heterothallic for sexual crossing. By breeding two of these mating-type engineered K. marxianus strains, we combined three complex traits—thermotolerance, lipid production, and facile transformation with exogenous DNA—into a single host. The ability to cross K. marxianus strains with relative ease, together with CRISPR-Cas9 genome editing, should enable engineering of K. marxianus isolates with promising lipid production at temperatures far exceeding those of other fungi under development for industrial applications. These results establish K. marxianus as a synthetic biology platform comparable to S. cerevisiae, with naturally more robust traits that hold potential for the industrial production of renewable chemicals. IMPORTANCE The yeast Kluyveromyces marxianus grows at high temperatures and on a wide range of carbon sources, making it a promising host for industrial biotechnology to produce renewable chemicals from plant biomass feedstocks. However, major genetic engineering limitations have kept this yeast from replacing the commonly used yeast Saccharomyces cerevisiae in industrial applications. Here, we describe genetic tools for genome editing and breeding K. marxianus strains, which we use to create a new thermotolerant strain with promising fatty acid production. These results open the door to using K. marxianus as a versatile synthetic biology platform organism for industrial applications.
format	article
author	Paul Cernak Raissa Estrela Snigdha Poddar Jeffrey M. Skerker Ya-Fang Cheng Annika K. Carlson Berling Chen Victoria M. Glynn Monique Furlan Owen W. Ryan Marie K. Donnelly Adam P. Arkin John W. Taylor Jamie H. D. Cate
author_facet	Paul Cernak Raissa Estrela Snigdha Poddar Jeffrey M. Skerker Ya-Fang Cheng Annika K. Carlson Berling Chen Victoria M. Glynn Monique Furlan Owen W. Ryan Marie K. Donnelly Adam P. Arkin John W. Taylor Jamie H. D. Cate
author_sort	Paul Cernak
title	Engineering <named-content content-type="genus-species">Kluyveromyces marxianus</named-content> as a Robust Synthetic Biology Platform Host
title_short	Engineering <named-content content-type="genus-species">Kluyveromyces marxianus</named-content> as a Robust Synthetic Biology Platform Host
title_full	Engineering <named-content content-type="genus-species">Kluyveromyces marxianus</named-content> as a Robust Synthetic Biology Platform Host
title_fullStr	Engineering <named-content content-type="genus-species">Kluyveromyces marxianus</named-content> as a Robust Synthetic Biology Platform Host
title_full_unstemmed	Engineering <named-content content-type="genus-species">Kluyveromyces marxianus</named-content> as a Robust Synthetic Biology Platform Host
title_sort	engineering <named-content content-type="genus-species">kluyveromyces marxianus</named-content> as a robust synthetic biology platform host
publisher	American Society for Microbiology
publishDate	2018
url	https://doaj.org/article/1ee3537f5483405a9dd2c47cbb421cca
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Engineering <named-content content-type="genus-species">Kluyveromyces marxianus</named-content> as a Robust Synthetic Biology Platform Host

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