Engineering increased triacylglycerol accumulation in Saccharomyces cerevisiae using a modified type 1 plant diacylglycerol acyltransferase

Verfasser / Beitragende:
[Michael Greer, Martin Truksa, Wei Deng, Shiu-Cheung Lung, Guanqun Chen, Randall Weselake]
Ort, Verlag, Jahr:
2015
Enthalten in:
Applied Microbiology and Biotechnology, 99/5(2015-03-01), 2243-2253
Format:
Artikel (online)
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024 7 0 |a 10.1007/s00253-014-6284-4  |2 doi 
035 |a (NATIONALLICENCE)springer-10.1007/s00253-014-6284-4 
245 0 0 |a Engineering increased triacylglycerol accumulation in Saccharomyces cerevisiae using a modified type 1 plant diacylglycerol acyltransferase  |h [Elektronische Daten]  |c [Michael Greer, Martin Truksa, Wei Deng, Shiu-Cheung Lung, Guanqun Chen, Randall Weselake] 
520 3 |a Diacylglycerol acyltransferase (DGAT) catalyzes the acyl-CoA-dependent acylation of sn-1,2-diacylglycerol to produce triacylglycerol (TAG). This enzyme, which is critical to numerous facets of oilseed development, has been highlighted as a genetic engineering target to increase storage lipid production in microorganisms designed for biofuel applications. Here, four transcriptionally active DGAT1 genes were identified and characterized from theoil crop Brassica napus. Overexpression of each BnaDGAT1 in Saccharomyces cerevisiae increased TAG biosynthesis. Further studies showed that adding an N-terminal tag could mask the deleterious influence of the DGATs' native N-terminal sequences, resulting in increased in vivo accumulation of the polypeptides and an increase of up to about 150-fold in in vitro enzyme activity. The levels of TAG and total lipid fatty acids in S. cerevisiae producing the N-terminally tagged BnaDGAT1.b at 72h were 53 and 28% higher than those in cultures producing untagged BnaA.DGAT1.b, respectively. These modified DGATs catalyzed the synthesis of up to 453mg fatty acid/L by this time point. The results will be of benefit in the biochemical analysis of recombinant DGAT1 produced through heterologous expression in yeast and offer a new approach to increase storage lipid content in yeast for industrial applications. 
540 |a Springer-Verlag Berlin Heidelberg, 2014 
690 7 |a Yeast  |2 nationallicence 
690 7 |a DGAT  |2 nationallicence 
690 7 |a Storage lipid synthesis  |2 nationallicence 
690 7 |a Brassica napus  |2 nationallicence 
690 7 |a Biofuel  |2 nationallicence 
700 1 |a Greer  |D Michael  |u Alberta Innovates Phytola Centre, Department of Agricultural, Food and Nutritional Science, University of Alberta, T6G 2P5, Edmonton, AB, Canada  |4 aut 
700 1 |a Truksa  |D Martin  |u Alberta Innovates Phytola Centre, Department of Agricultural, Food and Nutritional Science, University of Alberta, T6G 2P5, Edmonton, AB, Canada  |4 aut 
700 1 |a Deng  |D Wei  |u Alberta Innovates Phytola Centre, Department of Agricultural, Food and Nutritional Science, University of Alberta, T6G 2P5, Edmonton, AB, Canada  |4 aut 
700 1 |a Lung  |D Shiu-Cheung  |u Alberta Innovates Phytola Centre, Department of Agricultural, Food and Nutritional Science, University of Alberta, T6G 2P5, Edmonton, AB, Canada  |4 aut 
700 1 |a Chen  |D Guanqun  |u Alberta Innovates Phytola Centre, Department of Agricultural, Food and Nutritional Science, University of Alberta, T6G 2P5, Edmonton, AB, Canada  |4 aut 
700 1 |a Weselake  |D Randall  |u Alberta Innovates Phytola Centre, Department of Agricultural, Food and Nutritional Science, University of Alberta, T6G 2P5, Edmonton, AB, Canada  |4 aut 
773 0 |t Applied Microbiology and Biotechnology  |d Springer Berlin Heidelberg  |g 99/5(2015-03-01), 2243-2253  |x 0175-7598  |q 99:5<2243  |1 2015  |2 99  |o 253 
856 4 0 |u https://doi.org/10.1007/s00253-014-6284-4  |q text/html  |z Onlinezugriff via DOI 
898 |a BK010053  |b XK010053  |c XK010000 
900 7 |a Metadata rights reserved  |b Springer special CC-BY-NC licence  |2 nationallicence 
908 |D 1  |a research-article  |2 jats 
949 |B NATIONALLICENCE  |F NATIONALLICENCE  |b NL-springer 
950 |B NATIONALLICENCE  |P 856  |E 40  |u https://doi.org/10.1007/s00253-014-6284-4  |q text/html  |z Onlinezugriff via DOI 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Greer  |D Michael  |u Alberta Innovates Phytola Centre, Department of Agricultural, Food and Nutritional Science, University of Alberta, T6G 2P5, Edmonton, AB, Canada  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Truksa  |D Martin  |u Alberta Innovates Phytola Centre, Department of Agricultural, Food and Nutritional Science, University of Alberta, T6G 2P5, Edmonton, AB, Canada  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Deng  |D Wei  |u Alberta Innovates Phytola Centre, Department of Agricultural, Food and Nutritional Science, University of Alberta, T6G 2P5, Edmonton, AB, Canada  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Lung  |D Shiu-Cheung  |u Alberta Innovates Phytola Centre, Department of Agricultural, Food and Nutritional Science, University of Alberta, T6G 2P5, Edmonton, AB, Canada  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Chen  |D Guanqun  |u Alberta Innovates Phytola Centre, Department of Agricultural, Food and Nutritional Science, University of Alberta, T6G 2P5, Edmonton, AB, Canada  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Weselake  |D Randall  |u Alberta Innovates Phytola Centre, Department of Agricultural, Food and Nutritional Science, University of Alberta, T6G 2P5, Edmonton, AB, Canada  |4 aut 
950 |B NATIONALLICENCE  |P 773  |E 0-  |t Applied Microbiology and Biotechnology  |d Springer Berlin Heidelberg  |g 99/5(2015-03-01), 2243-2253  |x 0175-7598  |q 99:5<2243  |1 2015  |2 99  |o 253