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Applied Microbiology and Biotechnology

, Volume 74, Issue 1, pp 131–139 | Cite as

Engineering the lycopene synthetic pathway in E. coli by comparison of the carotenoid genes of Pantoea agglomerans and Pantoea ananatis

  • 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 KimEmail author
Applied Genetics and Molecular Biotechnology

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.

Keywords

Carotenoid Lycopene IPTG Induction crtE Lycopene Production 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

This work was supported by the BioGreen 21 Program (grant no.: 20050401034590) from the Korea Rural Development Administration, the Regional Innovation Program (grant no.: 10018072) from the Korea Ministry of Commerce, Industry and Energy, and a grant from the MOST/KOSEF (Environmental Biotechnology National Core Research Center) (grant no.: R15-2003-012-02001-0), Republic of Korea. Scholarships obtained by the second, third, and fourth authors were supported by the BK21 program of Korea. The authors Sang-Hwal Yoon and Ju-Eun Kim contributed equally to this work.

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Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Sang-Hwal Yoon
    • 2
  • Ju-Eun Kim
    • 1
  • Sook-Hee Lee
    • 1
  • Hye-Min Park
    • 1
  • Myung-Suk Choi
    • 2
    • 3
  • Jae-Yean Kim
    • 1
    • 2
  • Si-Hyoung Lee
    • 5
  • Yong-Chul Shin
    • 4
    • 5
  • Jay D. Keasling
    • 6
  • Seon-Won Kim
    • 1
    • 2
    Email author
  1. 1.Division of Applied Life Science (BK21)Gyeongsang National UniversityJinjuSouth Korea
  2. 2.Environmental Biotechnology National Core Research CenterGyeongsang National UniversityJinjuSouth Korea
  3. 3.Division of Forest ScienceGyeongsang National UniversityJinjuSouth Korea
  4. 4.Department of MicrobiologyGyeongsang National UniversityJinjuSouth Korea
  5. 5.Amicogen Inc.JinjuSouth Korea
  6. 6.Department of Chemical EngineeringUniversity of CaliforniaBerkeleyUSA

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