caa a22 4500 606213090 CHVBK 20210128101045.0 cr unu---uuuuu 210128e20150201xx s 000 0 eng 10.1007/s10295-014-1552-y doi (NATIONALLICENCE)springer-10.1007/s10295-014-1552-y The effects of putative lipase and wax ester synthase/acyl-CoA:diacylglycerol acyltransferase gene knockouts on triacylglycerol accumulation in Gordonia sp. KTR9 [Elektronische Daten] [Karl Indest, Jed Eberly, David Ringelberg, Dawn Hancock] Previously, we demonstrated triacylglycerol (TAG) accumulation and the in vivo ability to catalyze esters from exogenous short chain alcohol sources in Gordonia sp. strain KTR9. In this study, we investigated the effects that putative lipase (KTR9_0186) and wax ester synthase/acyl-CoA:diacylglycerol acyltransferase (WS/DGAT; KTR9_3844) gene knockouts had on TAG accumulation. Gene disruption of KTR9_0186 resulted in a twofold increase in TAG content in nitrogen starved cells. Lipase mutants subjected to carbon starvation, following nitrogen starvation, retained 75% more TAGs and retained pigmentation. Transcriptome expression data confirmed the deletion of KTR9_0186 and identified the up-regulation of key genes involved in fatty acid degradation, a likely compensatory mechanism for reduced TAG mobilization. In vitro assays with purified KTR9_3844 demonstrated WS/DGAT activity with short chain alcohols and C16 and C18 fatty acid Co-As. Collectively, these results indicate that Gordonia sp. KTR9 has a suitable tractable genetic background for TAG production as well as the enzymatic capacity to catalyze fatty acid esters from short chain alcohols. Springer-Verlag (outside the USA), 2014 Triacyglycerol nationallicence Lipase nationallicence WS/DGAT nationallicence Gordonia nationallicence Indest Karl Environmental Laboratory, CEERD EP-P, U.S. Army Engineer Research and Development Center, 3909 Halls Ferry Road, 39180, Vicksburg, MS, USA aut Eberly Jed Environmental Laboratory, CEERD EP-P, U.S. Army Engineer Research and Development Center, 3909 Halls Ferry Road, 39180, Vicksburg, MS, USA aut Ringelberg David Cold Regions Research and Engineering Laboratory, U.S. Army Engineer Research and Development Center, 03755, Hanover, NH, USA aut Hancock Dawn Environmental Laboratory, CEERD EP-P, U.S. Army Engineer Research and Development Center, 3909 Halls Ferry Road, 39180, Vicksburg, MS, USA aut Journal of Industrial Microbiology & Biotechnology Springer Berlin Heidelberg 42/2(2015-02-01), 219-227 1367-5435 42:2<219 2015 42 10295 https://doi.org/10.1007/s10295-014-1552-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/s10295-014-1552-y text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Indest Karl Environmental Laboratory, CEERD EP-P, U.S. Army Engineer Research and Development Center, 3909 Halls Ferry Road, 39180, Vicksburg, MS, USA aut NATIONALLICENCE 700 1- Eberly Jed Environmental Laboratory, CEERD EP-P, U.S. Army Engineer Research and Development Center, 3909 Halls Ferry Road, 39180, Vicksburg, MS, USA aut NATIONALLICENCE 700 1- Ringelberg David Cold Regions Research and Engineering Laboratory, U.S. Army Engineer Research and Development Center, 03755, Hanover, NH, USA aut NATIONALLICENCE 700 1- Hancock Dawn Environmental Laboratory, CEERD EP-P, U.S. Army Engineer Research and Development Center, 3909 Halls Ferry Road, 39180, Vicksburg, MS, USA aut NATIONALLICENCE 773 0- Journal of Industrial Microbiology & Biotechnology Springer Berlin Heidelberg 42/2(2015-02-01), 219-227 1367-5435 42:2<219 2015 42 10295