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Nannochloropsis, a rich source of diacylglycerol acyltransferases for engineering of triacylglycerol content in different hosts

BACKGROUND: Photosynthetic microalgae are considered a viable and sustainable resource for biofuel feedstocks, because they can produce higher biomass per land area than plants and can be grown on non-arable land. Among many microalgae considered for biofuel production, Nannochloropsis oceanica (CCM...

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Autores principales: Zienkiewicz, Krzysztof, Zienkiewicz, Agnieszka, Poliner, Eric, Du, Zhi-Yan, Vollheyde, Katharina, Herrfurth, Cornelia, Marmon, Sofia, Farré, Eva M., Feussner, Ivo, Benning, Christoph
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5210179/
https://www.ncbi.nlm.nih.gov/pubmed/28070221
http://dx.doi.org/10.1186/s13068-016-0686-8
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author Zienkiewicz, Krzysztof
Zienkiewicz, Agnieszka
Poliner, Eric
Du, Zhi-Yan
Vollheyde, Katharina
Herrfurth, Cornelia
Marmon, Sofia
Farré, Eva M.
Feussner, Ivo
Benning, Christoph
author_facet Zienkiewicz, Krzysztof
Zienkiewicz, Agnieszka
Poliner, Eric
Du, Zhi-Yan
Vollheyde, Katharina
Herrfurth, Cornelia
Marmon, Sofia
Farré, Eva M.
Feussner, Ivo
Benning, Christoph
author_sort Zienkiewicz, Krzysztof
collection PubMed
description BACKGROUND: Photosynthetic microalgae are considered a viable and sustainable resource for biofuel feedstocks, because they can produce higher biomass per land area than plants and can be grown on non-arable land. Among many microalgae considered for biofuel production, Nannochloropsis oceanica (CCMP1779) is particularly promising, because following nutrient deprivation it produces very high amounts of triacylglycerols (TAG). The committed step in TAG synthesis is catalyzed by acyl-CoA:diacylglycerol acyltransferase (DGAT). Remarkably, a total of 13 putative DGAT-encoding genes have been previously identified in CCMP1779 but most have not yet been studied in detail. RESULTS: Based on their expression profile, six out of 12 type-2 DGAT-encoding genes (NoDGTT1-NoDGTT6) were chosen for their possible role in TAG biosynthesis and the respective cDNAs were expressed in a TAG synthesis-deficient mutant of yeast. Yeast expressing NoDGTT5 accumulated TAG to the highest level. Over-expression of NoDGTT5 in CCMP1779 grown in N-replete medium resulted in levels of TAG normally observed only after N deprivation. Reduced growth rates accompanied NoDGTT5 over-expression in CCMP1779. Constitutive expression of NoDGTT5 in Arabidopsis thaliana was accompanied by increased TAG content in seeds and leaves. A broad substrate specificity for NoDGTT5 was revealed, with preference for unsaturated acyl groups. Furthermore, NoDGTT5 was able to successfully rescue the Arabidopsis tag1-1 mutant by restoring the TAG content in seeds. CONCLUSIONS: Taken together, our results identified NoDGTT5 as the most promising gene for the engineering of TAG synthesis in multiple hosts among the 13 DGAT-encoding genes of N. oceanica CCMP1779. Consequently, this study demonstrates the potential of NoDGTT5 as a tool for enhancing the energy density in biomass by increasing TAG content in transgenic crops used for biofuel production. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13068-016-0686-8) contains supplementary material, which is available to authorized users.
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spelling pubmed-52101792017-01-09 Nannochloropsis, a rich source of diacylglycerol acyltransferases for engineering of triacylglycerol content in different hosts Zienkiewicz, Krzysztof Zienkiewicz, Agnieszka Poliner, Eric Du, Zhi-Yan Vollheyde, Katharina Herrfurth, Cornelia Marmon, Sofia Farré, Eva M. Feussner, Ivo Benning, Christoph Biotechnol Biofuels Research BACKGROUND: Photosynthetic microalgae are considered a viable and sustainable resource for biofuel feedstocks, because they can produce higher biomass per land area than plants and can be grown on non-arable land. Among many microalgae considered for biofuel production, Nannochloropsis oceanica (CCMP1779) is particularly promising, because following nutrient deprivation it produces very high amounts of triacylglycerols (TAG). The committed step in TAG synthesis is catalyzed by acyl-CoA:diacylglycerol acyltransferase (DGAT). Remarkably, a total of 13 putative DGAT-encoding genes have been previously identified in CCMP1779 but most have not yet been studied in detail. RESULTS: Based on their expression profile, six out of 12 type-2 DGAT-encoding genes (NoDGTT1-NoDGTT6) were chosen for their possible role in TAG biosynthesis and the respective cDNAs were expressed in a TAG synthesis-deficient mutant of yeast. Yeast expressing NoDGTT5 accumulated TAG to the highest level. Over-expression of NoDGTT5 in CCMP1779 grown in N-replete medium resulted in levels of TAG normally observed only after N deprivation. Reduced growth rates accompanied NoDGTT5 over-expression in CCMP1779. Constitutive expression of NoDGTT5 in Arabidopsis thaliana was accompanied by increased TAG content in seeds and leaves. A broad substrate specificity for NoDGTT5 was revealed, with preference for unsaturated acyl groups. Furthermore, NoDGTT5 was able to successfully rescue the Arabidopsis tag1-1 mutant by restoring the TAG content in seeds. CONCLUSIONS: Taken together, our results identified NoDGTT5 as the most promising gene for the engineering of TAG synthesis in multiple hosts among the 13 DGAT-encoding genes of N. oceanica CCMP1779. Consequently, this study demonstrates the potential of NoDGTT5 as a tool for enhancing the energy density in biomass by increasing TAG content in transgenic crops used for biofuel production. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13068-016-0686-8) contains supplementary material, which is available to authorized users. BioMed Central 2017-01-03 /pmc/articles/PMC5210179/ /pubmed/28070221 http://dx.doi.org/10.1186/s13068-016-0686-8 Text en © The Author(s) 2017 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
Zienkiewicz, Krzysztof
Zienkiewicz, Agnieszka
Poliner, Eric
Du, Zhi-Yan
Vollheyde, Katharina
Herrfurth, Cornelia
Marmon, Sofia
Farré, Eva M.
Feussner, Ivo
Benning, Christoph
Nannochloropsis, a rich source of diacylglycerol acyltransferases for engineering of triacylglycerol content in different hosts
title Nannochloropsis, a rich source of diacylglycerol acyltransferases for engineering of triacylglycerol content in different hosts
title_full Nannochloropsis, a rich source of diacylglycerol acyltransferases for engineering of triacylglycerol content in different hosts
title_fullStr Nannochloropsis, a rich source of diacylglycerol acyltransferases for engineering of triacylglycerol content in different hosts
title_full_unstemmed Nannochloropsis, a rich source of diacylglycerol acyltransferases for engineering of triacylglycerol content in different hosts
title_short Nannochloropsis, a rich source of diacylglycerol acyltransferases for engineering of triacylglycerol content in different hosts
title_sort nannochloropsis, a rich source of diacylglycerol acyltransferases for engineering of triacylglycerol content in different hosts
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5210179/
https://www.ncbi.nlm.nih.gov/pubmed/28070221
http://dx.doi.org/10.1186/s13068-016-0686-8
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