Engineering the lycopene synthetic pathway in <Emphasis Type="Italic">E. coli</Emphasis> by comparison of the carotenoid genes of <Emphasis Type="Italic">Pantoea agglomerans</Emphasis> and <Emphasis Type="Italic">Pantoea ananatis</Emphasis> |
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Authors: | Sang-Hwal Yoon Ju-Eun Kim Sook-Hee Lee Hye-Min Park Myung-Suk Choi Jae-Yean Kim Si-Hyoung Lee Yong-Chul Shin Jay D Keasling Seon-Won Kim |
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Institution: | (1) Division of Applied Life Science (BK21), Gyeongsang National University, Jinju, 660-701, South Korea;(2) Environmental Biotechnology National Core Research Center, Gyeongsang National University, Jinju, 660-701, South Korea;(3) Division of Forest Science, Gyeongsang National University, Jinju, 660-701, South Korea;(4) Department of Microbiology, Gyeongsang National University, Jinju, 660-701, South Korea;(5) Amicogen Inc., Jinsung, Jinju, 660-852, South Korea;(6) Department of Chemical Engineering, University of California, Berkeley, CA 94720-1462, USA |
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Abstract: | The lycopene synthetic pathway was engineered in Escherichia coli using the carotenoid genes (crtE, crtB, and crtI) of Pantoea agglomerans and Pantoea ananatis. E. coli harboring the P. agglomerans crt genes produced 27 mg/l of lycopene in 2YT medium without isopropyl-beta-d-thiogalactopyranoside (IPTG) induction, which was twofold higher than that produced by E. coli harboring the P. ananatis crt genes (12 mg/l lycopene) with 0.1 mM IPTG induction. The crt genes of P. agglomerans proved better for lycopene production in E. coli than those of P. ananatis. The crt genes of the two bacteria were also compared in E. coli harboring the mevalonate bottom pathway, which was capable of providing sufficient carotenoid building blocks, isopentenyl
diphosphate (IPP) and dimethylallyl diphosphate (DMAPP), with exogenous mevalonate supplementation. Lycopene production significantly
increased using the mevalonate bottom pathway and 60 mg/l of lycopene was obtained with the P. agglomerans crt genes, which was higher than that obtained with the P. ananatis crt genes (35 mg/l lycopene). When crtE among the P. ananatis crt genes was replaced with P. agglomerans crtE or Archaeoglobus fulgidus gps, both lycopene production and cell growth were similar to that obtained with P. agglomerans
crt genes. The crtE gene was responsible for the observed difference in lycopene production and cell growth between E. coli harboring the crt genes of P. agglomerans and P. ananatis. As there was no significant difference in lycopene production between E. coli harboring P. agglomerans
crtE and A. fulgidus gps, farnesyl diphosphate (FPP) synthesis was not rate-limiting in E. coli.
Sang-Hwal Yoon and Ju-Eun Kim: These authors contributed equally to this work. |
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