caa a22 4500 606212515 CHVBK 20210128101043.0 cr unu---uuuuu 210128e20150301xx s 000 0 eng 10.1007/s10295-014-1540-2 doi (NATIONALLICENCE)springer-10.1007/s10295-014-1540-2 Metabolic pathway engineering for fatty acid ethyl ester production in Saccharomyces cerevisiae using stable chromosomal integration [Elektronische Daten] [Bouke de Jong, Shuobo Shi, Juan Valle-Rodríguez, Verena Siewers, Jens Nielsen] Fatty acid ethyl esters are fatty acid derived molecules similar to first generation biodiesel (fatty acid methyl esters; FAMEs) which can be produced in a microbial cell factory. Saccharomyces cerevisiae is a suitable candidate for microbial large scale and long term cultivations, which is the typical industrial production setting for biofuels. It is crucial to conserve the metabolic design of the cell factory during industrial cultivation conditions that require extensive propagation. Genetic modifications therefore have to be introduced in a stable manner. Here, several metabolic engineering strategies for improved production of fatty acid ethyl esters in S. cerevisiae were combined and the genes were stably expressed from the organisms' chromosomes. A wax ester synthase (ws2) was expressed in different yeast strains with an engineered acetyl-CoA and fatty acid metabolism. Thus, we compared expression of ws2 with and without overexpression of alcohol dehydrogenase (ADH2), acetaldehyde dehydrogenase (ALD6) and acetyl-CoA synthetase (acs SE L641P ) and further evaluated additional overexpression of a mutant version of acetyl-CoA decarboxylase (ACC1 S1157A,S659A ) and the acyl-CoA binding protein (ACB1). The combined engineering efforts of the implementation of ws2, ADH2, ALD6 and acs SE L641P , ACC1 S1157A,S659A and ACB1 in a S. cerevisiae strain lacking storage lipid formation (are1Δ, are2Δ, dga1Δ and lro1Δ) and β-oxidation (pox1Δ) resulted in a 4.1-fold improvement compared with sole expression of ws2 in S. cerevisiae. Society for Industrial Microbiology and Biotechnology, 2014 Yeast nationallicence Industrial strain nationallicence Chromosomal integration nationallicence Homologous recombination nationallicence Fatty acid ethyl ester (FAEE) nationallicence Metabolic pathway engineering nationallicence de Jong Bouke Department of Chemical and Biological Engineering, Chalmers University of Technology, 41296, Gothenburg, Sweden aut Shi Shuobo Institute of Chemical and Engineering Sciences, A-Star, 31, Biopolis Way, #01-01 Nanos, 138669, Singapore, Singapore aut Valle-Rodríguez Juan Department of Chemical and Biological Engineering, Chalmers University of Technology, 41296, Gothenburg, Sweden aut Siewers Verena Department of Chemical and Biological Engineering, Chalmers University of Technology, 41296, Gothenburg, Sweden aut Nielsen Jens Department of Chemical and Biological Engineering, Chalmers University of Technology, 41296, Gothenburg, Sweden aut Journal of Industrial Microbiology & Biotechnology Springer Berlin Heidelberg 42/3(2015-03-01), 477-486 1367-5435 42:3<477 2015 42 10295 https://doi.org/10.1007/s10295-014-1540-2 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/s10295-014-1540-2 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- de Jong Bouke Department of Chemical and Biological Engineering, Chalmers University of Technology, 41296, Gothenburg, Sweden aut NATIONALLICENCE 700 1- Shi Shuobo Institute of Chemical and Engineering Sciences, A-Star, 31, Biopolis Way, #01-01 Nanos, 138669, Singapore, Singapore aut NATIONALLICENCE 700 1- Valle-Rodríguez Juan Department of Chemical and Biological Engineering, Chalmers University of Technology, 41296, Gothenburg, Sweden aut NATIONALLICENCE 700 1- Siewers Verena Department of Chemical and Biological Engineering, Chalmers University of Technology, 41296, Gothenburg, Sweden aut NATIONALLICENCE 700 1- Nielsen Jens Department of Chemical and Biological Engineering, Chalmers University of Technology, 41296, Gothenburg, Sweden aut NATIONALLICENCE 773 0- Journal of Industrial Microbiology & Biotechnology Springer Berlin Heidelberg 42/3(2015-03-01), 477-486 1367-5435 42:3<477 2015 42 10295