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

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. We chose six out of 12 type-2 DGAT-encoding genes (NoDGTT1-NoDGTT6), based on their expression profile, 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 rescuemore » the Arabidopsis tag1-1 mutant by restoring the TAG content in seeds. Taken together, these 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.« less
Authors:
ORCiD logo [1] ;  [2] ;  [3] ;  [4] ;  [5] ;  [5] ;  [6] ;  [7] ;  [8] ;  [9]
  1. Michigan State Univ., East Lansing, MI (United States); Georg-August Univ., Gottingen (Germany). Albrecht-von-Haller-Inst. for Plant Sciences
  2. Michigan State Univ., East Lansing, MI (United States). Great Lakes Bioenergy Center; Georg-August Univ., Gottingen (Germany). Albrecht-von-Haller-Inst. for Plant Sciences
  3. Michigan State Univ., East Lansing, MI (United States). Cell and Molecular Biology Program
  4. Michigan State Univ., East Lansing, MI (United States)
  5. Georg-August Univ., Gottingen (Germany). Albrecht-von-Haller-Inst. for Plant Sciences
  6. Georg-August Univ., Gottingen (Germany). Albrecht-von-Haller-Inst. for Plant Sciences; Swedish Univ. of Agricultural Sciences (SLU), Alnarp (Sweden). Dept. of Plant Breeding
  7. Michigan State Univ., East Lansing, MI (United States). Dept. of Plant Biology
  8. Georg-August Univ., Gottingen (Germany). Albrecht-von-Haller-Inst. for Plant Sciences; Georg-August-University, Gottingen (Germany). Dept. of Plant Biochemistry, International Center for Advanced Studies Energy Conversion (ICASEC)
  9. Michigan State Univ., East Lansing, MI (United States). Great Lakes Bioenergy Center, Dept. of Plant Biology, Dept. of Biochemistry and Molecular Biology, Dept. of Plant Biology, Dept. of Biochemistry and Molecular Biology
Publication Date:
Grant/Contract Number:
FG02-91ER20021; FC02-07ER64494
Type:
Accepted Manuscript
Journal Name:
Biotechnology for Biofuels
Additional Journal Information:
Journal Volume: 10; Journal Issue: 1; Journal ID: ISSN 1754-6834
Publisher:
BioMed Central
Research Org:
Michigan State Univ., East Lansing, MI (United States); Univ. of Wisconsin, Madison, WI (United States)
Sponsoring Org:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; 09 BIOMASS FUELS; Nannochloropsis oceanica; Microalgae; DGAT; Triacylglycerol; Lipid storage; Lipid droplets
OSTI Identifier:
1366477

Zienkiewicz, Krzysztof, Zienkiewicz, Agnieszka, Poliner, Eric, Du, Zhi-Yan, Vollheyde, Katharina, Herrfurth, Cornelia, Marmon, Sofia, Farre, Eva M., Feussner, Ivo, and Benning, Christoph. Nannochloropsis, a rich source of diacylglycerol acyltransferases for engineering of triacylglycerol content in different hosts. United States: N. p., Web. doi:10.1186/s13068-016-0686-8.
Zienkiewicz, Krzysztof, Zienkiewicz, Agnieszka, Poliner, Eric, Du, Zhi-Yan, Vollheyde, Katharina, Herrfurth, Cornelia, Marmon, Sofia, Farre, Eva M., Feussner, Ivo, & Benning, Christoph. Nannochloropsis, a rich source of diacylglycerol acyltransferases for engineering of triacylglycerol content in different hosts. United States. doi:10.1186/s13068-016-0686-8.
Zienkiewicz, Krzysztof, Zienkiewicz, Agnieszka, Poliner, Eric, Du, Zhi-Yan, Vollheyde, Katharina, Herrfurth, Cornelia, Marmon, Sofia, Farre, Eva M., Feussner, Ivo, and Benning, Christoph. 2017. "Nannochloropsis, a rich source of diacylglycerol acyltransferases for engineering of triacylglycerol content in different hosts". United States. doi:10.1186/s13068-016-0686-8. https://www.osti.gov/servlets/purl/1366477.
@article{osti_1366477,
title = {Nannochloropsis, a rich source of diacylglycerol acyltransferases for engineering of triacylglycerol content in different hosts},
author = {Zienkiewicz, Krzysztof and Zienkiewicz, Agnieszka and Poliner, Eric and Du, Zhi-Yan and Vollheyde, Katharina and Herrfurth, Cornelia and Marmon, Sofia and Farre, Eva M. and Feussner, Ivo and Benning, Christoph},
abstractNote = {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. We chose six out of 12 type-2 DGAT-encoding genes (NoDGTT1-NoDGTT6), based on their expression profile, 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. Taken together, these 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.},
doi = {10.1186/s13068-016-0686-8},
journal = {Biotechnology for Biofuels},
number = 1,
volume = 10,
place = {United States},
year = {2017},
month = {1}
}

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