caa a22 4500 605500959 CHVBK 20210128100558.0 cr unu---uuuuu 210128e20151101xx s 000 0 eng 10.1007/s00253-015-6783-y doi (NATIONALLICENCE)springer-10.1007/s00253-015-6783-y Altering small and medium alcohol selectivity in the wax ester synthase [Elektronische Daten] [Brett Barney, Janet Ohlert, Jacobe Timler, Amelia Lijewski] The bifunctional wax ester synthase/acyl-coenzyme A:diacylglycerol acyltransferase (WS/DGAT or wax ester synthase) catalyzes the terminal reaction in the bacterial wax ester biosynthetic pathway, utilizing a range of alcohols and fatty acyl-CoAs to synthesize the corresponding wax ester. The wild-type wax ester synthase Maqu_0168 from Marinobacter aquaeolei VT8 exhibits a preference for longer fatty alcohols, while applications with smaller alcohols would yield products with desired biotechnological properties. Small and medium chain length alcohol substrates are much poorer substrates for the native enzyme, which may hinder broad application of the wax ester synthase in many proposed biosynthetic schemes. Developing approaches to improve enzyme activity toward specific smaller alcohol substrates first requires a clear understanding of which amino acids of the primary sequences of these enzymes contribute to substrate specificity in the native enzyme. In this report, we surveyed a range of potential residues and identified the leucine at position 356 and methionine at position 405 in Maqu_0168 as residues that affected selectivity toward small, branched, and aromatic alcohols when substituted with different amino acids. This analysis provides evidence of residues that line the binding site for wax ester synthase, which will aid rational approaches to improve this enzyme with specific substrates. Springer-Verlag Berlin Heidelberg, 2015 Wax ester synthase nationallicence WS/DGAT nationallicence Ethanol nationallicence Isoamyl alcohol nationallicence Fatty acyl-CoA nationallicence Barney Brett Department of Bioproducts and Biosystems Engineering and BioTechnology Institute, University of Minnesota, 1390 Eckles Avenue, 55108-6130, St. Paul, MN, USA aut Ohlert Janet Department of Bioproducts and Biosystems Engineering and BioTechnology Institute, University of Minnesota, 1390 Eckles Avenue, 55108-6130, St. Paul, MN, USA aut Timler Jacobe Department of Bioproducts and Biosystems Engineering and BioTechnology Institute, University of Minnesota, 1390 Eckles Avenue, 55108-6130, St. Paul, MN, USA aut Lijewski Amelia Department of Bioproducts and Biosystems Engineering and BioTechnology Institute, University of Minnesota, 1390 Eckles Avenue, 55108-6130, St. Paul, MN, USA aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/22(2015-11-01), 9675-9684 0175-7598 99:22<9675 2015 99 253 https://doi.org/10.1007/s00253-015-6783-y 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/s00253-015-6783-y text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Barney Brett Department of Bioproducts and Biosystems Engineering and BioTechnology Institute, University of Minnesota, 1390 Eckles Avenue, 55108-6130, St. Paul, MN, USA aut NATIONALLICENCE 700 1- Ohlert Janet Department of Bioproducts and Biosystems Engineering and BioTechnology Institute, University of Minnesota, 1390 Eckles Avenue, 55108-6130, St. Paul, MN, USA aut NATIONALLICENCE 700 1- Timler Jacobe Department of Bioproducts and Biosystems Engineering and BioTechnology Institute, University of Minnesota, 1390 Eckles Avenue, 55108-6130, St. Paul, MN, USA aut NATIONALLICENCE 700 1- Lijewski Amelia Department of Bioproducts and Biosystems Engineering and BioTechnology Institute, University of Minnesota, 1390 Eckles Avenue, 55108-6130, St. Paul, MN, USA aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/22(2015-11-01), 9675-9684 0175-7598 99:22<9675 2015 99 253