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The effects of putative lipase and wax ester synthase/acyl-CoA:diacylglycerol acyltransferase gene knockouts on triacylglycerol accumulation in Gordonia sp. KTR9

  • Bioenergy/Biofuels/Biochemicals
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

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.

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Acknowledgments

This research was funded through the US Army Engineer Research and Development Center's Basic Research Program (Project 10–50, K. J. Indest). Views, opinions and/or findings contained herein are those of the authors and should not be construed as an official Department of the Army position or decision unless so designated by other official documentation.

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Authors and Affiliations

  1. Environmental Laboratory, CEERD EP-P, U.S. Army Engineer Research and Development Center, 3909 Halls Ferry Road, Vicksburg, MS, 39180, USA

    Karl J. Indest, Jed O. Eberly & Dawn E. Hancock

  2. Cold Regions Research and Engineering Laboratory, U.S. Army Engineer Research and Development Center, Hanover, NH, 03755, USA

    David B. Ringelberg

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  1. Karl J. Indest
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  2. Jed O. Eberly
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  3. David B. Ringelberg
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  4. Dawn E. Hancock
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Correspondence to Karl J. Indest.

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Indest, K.J., Eberly, J.O., Ringelberg, D.B. et al. The effects of putative lipase and wax ester synthase/acyl-CoA:diacylglycerol acyltransferase gene knockouts on triacylglycerol accumulation in Gordonia sp. KTR9. J Ind Microbiol Biotechnol 42, 219–227 (2015). https://doi.org/10.1007/s10295-014-1552-y

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