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Applied and Environmental Microbiology, February 2006, p. 1373-1379, Vol. 72, No. 2
0099-2240/06/$08.00+0     doi:10.1128/AEM.72.2.1373-1379.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Neutral Lipid Biosynthesis in Engineered Escherichia coli: Jojoba Oil-Like Wax Esters and Fatty Acid Butyl Esters

Rainer Kalscheuer,^1, Tim Stöveken,¹ Heinrich Luftmann,² Ursula Malkus,³ Rudolf Reichelt,³ and Alexander Steinbüchel^1*

Institut für Molekulare Mikrobiologie und Biotechnologie,¹ Institut für Organische Chemie,² Institut für Medizinische Physik und Biophysik, Universitätsklinikum, Westfälische Wilhelms-Universität, Münster, Germany³

Received 28 August 2005/ Accepted 22 November 2005

Wax esters are esters of long-chain fatty acids and long-chain fatty alcohols which are of considerable commercial importance and are produced on a scale of 3 million tons per year. The oil from the jojoba plant (Simmondsia chinensis) is the main biological source of wax esters. Although it has a multitude of potential applications, the use of jojoba oil is restricted, due to its high price. In this study, we describe the establishment of heterologous wax ester biosynthesis in a recombinant Escherichia coli strain by coexpression of a fatty alcohol-producing bifunctional acyl-coenzyme A reductase from the jojoba plant and a bacterial wax ester synthase from Acinetobacter baylyi strain ADP1, catalyzing the esterification of fatty alcohols and coenzyme A thioesters of fatty acids. In the presence of oleate, jojoba oil-like wax esters such as palmityl oleate, palmityl palmitoleate, and oleyl oleate were produced, amounting to up to ca. 1% of the cellular dry weight. In addition to wax esters, fatty acid butyl esters were unexpectedly observed in the presence of oleate. The latter could be attributed to solvent residues of 1-butanol present in the medium component, Bacto tryptone. Neutral lipids produced in recombinant E. coli were accumulated as intracytoplasmic inclusions, demonstrating that the formation and structural integrity of bacterial lipid bodies do not require specific structural proteins. This is the first report on substantial biosynthesis and accumulation of neutral lipids in E. coli, which might open new perspectives for the biotechnological production of cheap jojoba oil equivalents from inexpensive resources employing recombinant microorganisms.

Present address: Department of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, NY 10461.

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