In vitro synthesis of 9,10-dihydroxyhexadecanoic acid using recombinant Escherichia coli

Microbial Cell Factories

Research

Preeti Srivastava¹  Virendra Swarup Bisaria¹  Anees Kaprakkaden² 

[1] Department of Biochemical Engineering and Biotechnology, Indian Institute of Technology Delhi, New Delhi, India;Department of Biochemical Engineering and Biotechnology, Indian Institute of Technology Delhi, New Delhi, India;Lac Production Division, ICAR-IINRG, Ranchi, India;

关键词: Fatty acid desaturase;    Epoxygenase;    Epoxide hydrolase;    Fatty acid hydroxylation;    E. coli;   

DOI  :  10.1186/s12934-017-0696-7

 received in 2016-10-22, accepted in 2017-05-05,  发布年份 2017

来源: Springer

【 摘 要 】

BackgroundHydroxy fatty acids are widely used in food, chemical and cosmetic industries. A variety of dihydroxy fatty acids have been synthesized so far; however, no studies have been done on the synthesis of 9,10-dihydroxyhexadecanoic acid. In the present study recombinant E. coli has been used for the heterologous expression of fatty acid hydroxylating enzymes and the whole cell lysate of the induced culture was used for in vitro production of 9,10-dihydroxyhexadecanoic acid.ResultsA first of its kind proof of principle has been successfully demonstrated for the production of 9,10-dihydroxyhexadecanoic acid using three different enzymes viz. fatty acid desaturase (FAD) from Saccharomyces cerevisiae, epoxide hydrolase (EH) from Caenorhabditis elegance and epoxygenase (EPOX) from Stokasia laevis. The genes for these proteins were codon-optimised, synthesised and cloned in pET 28a (+) vector. The culture conditions for induction of these three proteins in E. coli were optimised in shake flask. The induced cell lysates were used both singly and in combination along with the trans-supply of hexadecanoic acid and 9-hexadecenoic acid, followed by product profiling by GC–MS. Formation of 9,10-dihydroxyhexadecanoic acid was successfully achieved when combination of induced cell lysates of recombinant E. coli containing FAD, EH, and EPOX were incubated with 9-hexadecenoic acid.ConclusionsThe in vitro production of 9,10-dihydroxyhexadecanoic acid synthesis using three fatty acid modification genes from different sources has been successfully demonstrated. The strategy adopted can be used for the production of similar compounds.

【 授权许可】

CC BY   
© The Author(s) 2017