Conflict of interest: none.
Systems metabolic engineering of Escherichia coli for gram scale production of the antitumor drug deoxyviolacein from glycerol
Article first published online: 25 AUG 2014
© 2014 Wiley Periodicals, Inc.
Biotechnology and Bioengineering
Volume 111, Issue 11, pages 2280–2289, November 2014
How to Cite
Rodrigues, A. L., Becker, J., de Souza Lima, A. O., Porto, L. M. and Wittmann, C. (2014), Systems metabolic engineering of Escherichia coli for gram scale production of the antitumor drug deoxyviolacein from glycerol. Biotechnol. Bioeng., 111: 2280–2289. doi: 10.1002/bit.25297
- Issue published online: 23 SEP 2014
- Article first published online: 25 AUG 2014
- Accepted manuscript online: 2 JUN 2014 03:27PM EST
- Manuscript Accepted: 21 MAY 2014
- Manuscript Revised: 24 APR 2014
- Manuscript Received: 17 MAR 2014
- German Academic Exchange Service A/08/71658
- Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
- Max-Buchner Foundation of the DECHEMA KZ2924
- Conselho Nacional de Desenvolvimento Científico e Tecnológico - CNPq, Brazil
- 13C flux;
- natural product;
Deoxyviolacein is a microbial drug with biological activity against tumors, gram-positive bacteria, and fungal plant pathogens. Here, we describe an Escherichia coli strain for heterologous production of this high-value drug from glycerol. Plasmid-based expression of the deoxyviolacein cluster vioABCE was controlled by the araBAD promoter and induction by L-arabinose. Through elimination of L-arabinose catabolism in E. coli, the pentose sugar could be fully directed to induction of deoxyviolacein biosynthesis and was no longer metabolized, as verified by 13C isotope experiments. Deletion of the araBAD genes beneficially complemented with previously described (i) engineering of the pentose phosphate pathway, (ii) chorismate biosynthesis, (iii) tryptophan biosynthesis, (iv) improved supply of L-serine, (v) elimination of tryptophan repression, and (vi) of tryptophan catabolism. Subsequent screening of the created next-generation producer E. coli dVio-8 identified glycerol as optimum carbon source and a level of 100 mg L−1 of L-arabinose as optimum for induction. Transferred to a glycerol-based fed-batch process, E. coli dVio-8 surpassed the gram scale and produced 1.6 g L−1 deoxyviolacein. With straightforward extraction from culture broth and purification by flash chromatography, deoxyviolacein was obtained at >99.5% purity. Biotechnol. Bioeng. 2014;111: 2280–2289. © 2014 Wiley Periodicals, Inc.