Publication: Lycopene overproduction and in situ extraction in organic-aqueous culture systems using a metabolically engineered Escherichia coli
Authors
De Diego Puente, María Teresa ; Gallego Jara, Julia ; Del Real, Álvaro ; Écija Conesa, Ana ; Manjón Rubio, Arturo ; Cánovas Diaz, Manuel
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Description
©2015. This manuscript version is made available under the CC-BY 4.0 license http://creativecommons.org/licenses/by /4.0/
This document is the Published, version of a Published Work that appeared in final form in AMB Express. To access the final edited and published work see https://doi.org/10.1186/s13568-015-0150-3
Abstract
Lycopene is an important compound with an increasing industrial value. However, there is still no biotechnological
process to obtain it. In this study, a semi-continuous system for lycopene extraction from recombinant Escherichia
coli BL21 cells is proposed. A two-phase culture mode using organic solvents was found to maximize lycopene
production through in situ extraction from cells. Within the reactor, three phases were formed during the process: an
aqueous phase containing the recombinant E. coli, an interphase, and an organic phase. Lycopene was extracted from
the cells to both the interphase and the organic phase and, consequently, thus enhancing its production. Maximum
lycopene production (74.71 ± 3.74 mg L−1
) was obtained for an octane-aqueous culture system using the E. coli
BL21LF strain, a process that doubled the level obtained in the control aqueous culture. Study of the interphase by
transmission electron microscopy (TEM) showed the proteo-lipidic nature and the high storage capacity of lyco‑
pene. Moreover, a cell viability test by flow cytometry (CF) after 24 h of culture indicated that 24 % of the population
could be re-used. Therefore, a batch series reactor was designed for semi-continuous lycopene extraction. After five
cycles of operation (120 h), lycopene production was similar to that obtained in the control aqueous medium. A final
specific lycopene yield of up to 49.70 ± 2.48 mg g−1
was reached at 24 h, which represents to the highest titer to
date. In conclusion, the aqueous-organic semi-continuous culture system proposed is the first designed for lycopene
extraction, representing an important breakthrough in the development of a competitive biotechnological process
for lycopene production and extraction.
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