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Engineering of a high lipid producing Yarrowia lipolytica strain

BACKGROUND: Microbial lipids are produced by many oleaginous organisms including the well-characterized yeast Yarrowia lipolytica, which can be engineered for increased lipid yield by up-regulation of the lipid biosynthetic pathway and down-regulation or deletion of competing pathways. RESULTS: We d...

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Autores principales: Friedlander, Jonathan, Tsakraklides, Vasiliki, Kamineni, Annapurna, Greenhagen, Emily H., Consiglio, Andrew L., MacEwen, Kyle, Crabtree, Donald V., Afshar, Jonathan, Nugent, Rebecca L., Hamilton, Maureen A., Joe Shaw, A., South, Colin R., Stephanopoulos, Gregory, Brevnova, Elena E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4815080/
https://www.ncbi.nlm.nih.gov/pubmed/27034715
http://dx.doi.org/10.1186/s13068-016-0492-3
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author Friedlander, Jonathan
Tsakraklides, Vasiliki
Kamineni, Annapurna
Greenhagen, Emily H.
Consiglio, Andrew L.
MacEwen, Kyle
Crabtree, Donald V.
Afshar, Jonathan
Nugent, Rebecca L.
Hamilton, Maureen A.
Joe Shaw, A.
South, Colin R.
Stephanopoulos, Gregory
Brevnova, Elena E.
author_facet Friedlander, Jonathan
Tsakraklides, Vasiliki
Kamineni, Annapurna
Greenhagen, Emily H.
Consiglio, Andrew L.
MacEwen, Kyle
Crabtree, Donald V.
Afshar, Jonathan
Nugent, Rebecca L.
Hamilton, Maureen A.
Joe Shaw, A.
South, Colin R.
Stephanopoulos, Gregory
Brevnova, Elena E.
author_sort Friedlander, Jonathan
collection PubMed
description BACKGROUND: Microbial lipids are produced by many oleaginous organisms including the well-characterized yeast Yarrowia lipolytica, which can be engineered for increased lipid yield by up-regulation of the lipid biosynthetic pathway and down-regulation or deletion of competing pathways. RESULTS: We describe a strain engineering strategy centered on diacylglycerol acyltransferase (DGA) gene overexpression that applied combinatorial screening of overexpression and deletion genetic targets to construct a high lipid producing yeast biocatalyst. The resulting strain, NS432, combines overexpression of a heterologous DGA1 enzyme from Rhodosporidium toruloides, a heterlogous DGA2 enzyme from Claviceps purpurea, and deletion of the native TGL3 lipase regulator. These three genetic modifications, selected for their effect on lipid production, enabled a 77 % lipid content and 0.21 g lipid per g glucose yield in batch fermentation. In fed-batch glucose fermentation NS432 produced 85 g/L lipid at a productivity of 0.73 g/L/h. CONCLUSIONS: The yields, productivities, and titers reported in this study may further support the applied goal of cost-effective, large -scale microbial lipid production for use as biofuels and biochemicals. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13068-016-0492-3) contains supplementary material, which is available to authorized users.
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spelling pubmed-48150802016-04-01 Engineering of a high lipid producing Yarrowia lipolytica strain Friedlander, Jonathan Tsakraklides, Vasiliki Kamineni, Annapurna Greenhagen, Emily H. Consiglio, Andrew L. MacEwen, Kyle Crabtree, Donald V. Afshar, Jonathan Nugent, Rebecca L. Hamilton, Maureen A. Joe Shaw, A. South, Colin R. Stephanopoulos, Gregory Brevnova, Elena E. Biotechnol Biofuels Research BACKGROUND: Microbial lipids are produced by many oleaginous organisms including the well-characterized yeast Yarrowia lipolytica, which can be engineered for increased lipid yield by up-regulation of the lipid biosynthetic pathway and down-regulation or deletion of competing pathways. RESULTS: We describe a strain engineering strategy centered on diacylglycerol acyltransferase (DGA) gene overexpression that applied combinatorial screening of overexpression and deletion genetic targets to construct a high lipid producing yeast biocatalyst. The resulting strain, NS432, combines overexpression of a heterologous DGA1 enzyme from Rhodosporidium toruloides, a heterlogous DGA2 enzyme from Claviceps purpurea, and deletion of the native TGL3 lipase regulator. These three genetic modifications, selected for their effect on lipid production, enabled a 77 % lipid content and 0.21 g lipid per g glucose yield in batch fermentation. In fed-batch glucose fermentation NS432 produced 85 g/L lipid at a productivity of 0.73 g/L/h. CONCLUSIONS: The yields, productivities, and titers reported in this study may further support the applied goal of cost-effective, large -scale microbial lipid production for use as biofuels and biochemicals. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13068-016-0492-3) contains supplementary material, which is available to authorized users. BioMed Central 2016-03-31 /pmc/articles/PMC4815080/ /pubmed/27034715 http://dx.doi.org/10.1186/s13068-016-0492-3 Text en © Friedlander et al. 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Friedlander, Jonathan
Tsakraklides, Vasiliki
Kamineni, Annapurna
Greenhagen, Emily H.
Consiglio, Andrew L.
MacEwen, Kyle
Crabtree, Donald V.
Afshar, Jonathan
Nugent, Rebecca L.
Hamilton, Maureen A.
Joe Shaw, A.
South, Colin R.
Stephanopoulos, Gregory
Brevnova, Elena E.
Engineering of a high lipid producing Yarrowia lipolytica strain
title Engineering of a high lipid producing Yarrowia lipolytica strain
title_full Engineering of a high lipid producing Yarrowia lipolytica strain
title_fullStr Engineering of a high lipid producing Yarrowia lipolytica strain
title_full_unstemmed Engineering of a high lipid producing Yarrowia lipolytica strain
title_short Engineering of a high lipid producing Yarrowia lipolytica strain
title_sort engineering of a high lipid producing yarrowia lipolytica strain
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4815080/
https://www.ncbi.nlm.nih.gov/pubmed/27034715
http://dx.doi.org/10.1186/s13068-016-0492-3
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