caa a22 4500 60550444X CHVBK 20210128100616.0 cr unu---uuuuu 210128e20150301xx s 000 0 eng 10.1007/s00253-014-6284-4 doi (NATIONALLICENCE)springer-10.1007/s00253-014-6284-4 Engineering increased triacylglycerol accumulation in Saccharomyces cerevisiae using a modified type 1 plant diacylglycerol acyltransferase [Elektronische Daten] [Michael Greer, Martin Truksa, Wei Deng, Shiu-Cheung Lung, Guanqun Chen, Randall Weselake] 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. Springer-Verlag Berlin Heidelberg, 2014 Yeast nationallicence DGAT nationallicence Storage lipid synthesis nationallicence Brassica napus nationallicence Biofuel nationallicence Greer Michael Alberta Innovates Phytola Centre, Department of Agricultural, Food and Nutritional Science, University of Alberta, T6G 2P5, Edmonton, AB, Canada aut Truksa Martin Alberta Innovates Phytola Centre, Department of Agricultural, Food and Nutritional Science, University of Alberta, T6G 2P5, Edmonton, AB, Canada aut Deng Wei Alberta Innovates Phytola Centre, Department of Agricultural, Food and Nutritional Science, University of Alberta, T6G 2P5, Edmonton, AB, Canada aut Lung Shiu-Cheung Alberta Innovates Phytola Centre, Department of Agricultural, Food and Nutritional Science, University of Alberta, T6G 2P5, Edmonton, AB, Canada aut Chen Guanqun Alberta Innovates Phytola Centre, Department of Agricultural, Food and Nutritional Science, University of Alberta, T6G 2P5, Edmonton, AB, Canada aut Weselake Randall Alberta Innovates Phytola Centre, Department of Agricultural, Food and Nutritional Science, University of Alberta, T6G 2P5, Edmonton, AB, Canada aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/5(2015-03-01), 2243-2253 0175-7598 99:5<2243 2015 99 253 https://doi.org/10.1007/s00253-014-6284-4 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-014-6284-4 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Greer Michael Alberta Innovates Phytola Centre, Department of Agricultural, Food and Nutritional Science, University of Alberta, T6G 2P5, Edmonton, AB, Canada aut NATIONALLICENCE 700 1- Truksa Martin Alberta Innovates Phytola Centre, Department of Agricultural, Food and Nutritional Science, University of Alberta, T6G 2P5, Edmonton, AB, Canada aut NATIONALLICENCE 700 1- Deng Wei Alberta Innovates Phytola Centre, Department of Agricultural, Food and Nutritional Science, University of Alberta, T6G 2P5, Edmonton, AB, Canada aut NATIONALLICENCE 700 1- Lung Shiu-Cheung Alberta Innovates Phytola Centre, Department of Agricultural, Food and Nutritional Science, University of Alberta, T6G 2P5, Edmonton, AB, Canada aut NATIONALLICENCE 700 1- Chen Guanqun Alberta Innovates Phytola Centre, Department of Agricultural, Food and Nutritional Science, University of Alberta, T6G 2P5, Edmonton, AB, Canada aut NATIONALLICENCE 700 1- Weselake Randall Alberta Innovates Phytola Centre, Department of Agricultural, Food and Nutritional Science, University of Alberta, T6G 2P5, Edmonton, AB, Canada aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/5(2015-03-01), 2243-2253 0175-7598 99:5<2243 2015 99 253