3 Biotech

, 8:269 | Cite as

Engineering membrane morphology and manipulating synthesis for increased lycopene accumulation in Escherichia coli cell factories

  • Tao Wu
  • Lijun Ye
  • Dongdong Zhao
  • Siwei Li
  • Qingyan Li
  • Bolin Zhang
  • Changhao Bi
Original Article


The goal of this work was to improve the lycopene storage capacity of the E. coli membrane by engineering both morphological and biosynthetic aspects. First, Almgs, a protein from Acholeplasma laidlawii that is involved in membrane bending is overexpressed to expand the storage space for lycopene, which resulted in a 12% increase of specific lycopene production. Second, several genes related to the membrane-synthesis pathway in E. coli, including plsb, plsc, and dgka, were also overexpressed, which led to a further 13% increase. In addition, membrane separation and component analysis confirmed that the increased amount of lycopene was mainly accumulated within the cell membranes. Finally, by integrating both aforementioned modification strategies, a synergistic effect could be observed which caused a 1.32-fold increase of specific lycopene production, from the 27.5 mg/g of the parent to 36.4 mg/g DCW in the engineered strain. This work demonstrates that membrane engineering is a feasible strategy for increasing the production and accumulation of lycopene in E. coli.


Membrane morphology Membrane synthesis Lycopene Production Escherichia coli 



This research was financially supported by the National Natural Science Foundation of China (31522002), National High Technology Research and Development Program of China (2015AA020202), Tianjin Key Technology R&D program of Tianjin Municipal Science and Technology Commission (14ZCZDSY00067), and Novo Nordisk-Chinese Academy of Sciences (NN-CAS) Research Fund (NN-CAS-2015-2).

Author Contributions

WT performed research, analyzed data, designed research, and wrote the paper; YL, ZD, and LS designed research and analyzed data; LQ provided the bacteria; BC and ZB designed research, analyzed data, and wrote the paper. All authors read and approved the final manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare no competing financial interests.

Supplementary material

13205_2018_1298_MOESM1_ESM.docx (327 kb)
Supplementary material 1 (DOCX 326 KB)
13205_2018_1298_MOESM2_ESM.docx (641 kb)
Supplementary material 2 (DOCX 641 KB)


  1. Ahrazem O, Rubio-Moraga A, Berman J, Capell T, Christou P, Zhu C, Gomez-Gomez L (2016) The carotenoid cleavage dioxygenase CCD2 catalysing the synthesis of crocetin in spring crocuses and saffron is a plastidial enzyme. New Phytol 209(2):650–663. CrossRefPubMedGoogle Scholar
  2. Ajikumar PK, Tyo K, Carlsen S, Mucha O, Phon TH, Stephanopoulos G (2008) Terpenoids: opportunities for biosynthesis of natural product drugs using engineered microorganisms. Mol Pharm 5(2):167CrossRefPubMedGoogle Scholar
  3. Alper H, Jin YS, Moxley JF, Stephanopoulos G (2005) Identifying gene targets for the metabolic engineering of lycopene biosynthesis in Escherichia coli. Metab Eng 7(3):155–164CrossRefPubMedGoogle Scholar
  4. Bhataya A, Schmidt-Dannert C, Lee PC (2009) Metabolic engineering of Pichia pastoris X-33 for lycopene production. Process Biochem 44(10):1095–1102CrossRefGoogle Scholar
  5. Choi HS, Lee SY, Kim TY, Woo HM (2010) In silico identification of gene amplification targets for improvement of lycopene production. Appl Environ Microbiol 76(10):3097–3105. CrossRefPubMedPubMedCentralGoogle Scholar
  6. Clinton SK (1998) Lycopene: chemistry, biology, and implications for human health and disease. Nutr Rev 56(2):35–51CrossRefPubMedGoogle Scholar
  7. Cronan JE Jr, Rock CO (2008) Biosynthesis of membrane lipids. EcoSal Plus. CrossRefPubMedGoogle Scholar
  8. Eriksson HM, Wessman P, Ge C, Edwards K, Wieslander A (2009) Massive formation of intracellular membrane vesicles in Escherichia coli by a monotopic membrane-bound lipid glycosyltransferase. J Biol Chem 284(49):33904–33914. CrossRefPubMedPubMedCentralGoogle Scholar
  9. Farmer WR, Liao JC (2000) Improving lycopene production in Escherichia coli by engineering metabolic control. Nat Biotechnol 18(5):533–537CrossRefPubMedGoogle Scholar
  10. Herskovits AA, Shimoni E, Minsky A, Bibi E (2002) Accumulation of endoplasmic membranes and novel membrane-bound ribosome-signal recognition particle receptor complexes in Escherichia coli. J Cell Biol 159(3):403–410. CrossRefPubMedPubMedCentralGoogle Scholar
  11. Hillson NJ, Rosengarten RD, Keasling JD (2012) j5 DNA assembly design automation software. ACS Synth Biol 1(1):14–21. CrossRefPubMedGoogle Scholar
  12. Janßen HJ, Steinbüchel A (2014) Fatty acid synthesis in Escherichia coli and its applications towards the production of fatty acid based biofuels. Biotechnol Biofuels 7(1):7CrossRefPubMedPubMedCentralGoogle Scholar
  13. Jin YS, Stephanopoulos G (2007) Multi-dimensional gene target search for improving lycopene biosynthesis in Escherichia coli. Metab Eng 9(4):337–347. CrossRefPubMedGoogle Scholar
  14. Kang MJ, Lee YM, Yoon SH, Kim JH, Ock SW, Jung KH, Shin YC, Keasling JD, Kim SW (2005) Identification of genes affecting lycopene accumulation in Escherichia coli using a shot-gun method. Biotechnol Bioeng 91(5):636CrossRefPubMedGoogle Scholar
  15. Kim YS, Lee JH, Kim NH, Yeom SJ, Kim SW, Oh DK (2011) Increase of lycopene production by supplementing auxiliary carbon sources in metabolically engineered Escherichia coli. Appl Microbiol Biotechnol 90(2):489–497CrossRefPubMedGoogle Scholar
  16. Lu J, Tang J, Liu Y, Zhu X, Zhang T, Zhang X (2012) Combinatorial modulation of galP and glk gene expression for improved alternative glucose utilization. Appl Microbiol Biotechnol 93(6):2455–2462. CrossRefPubMedGoogle Scholar
  17. Lu P, Ma D, Yan C, Gong X, Du M, Shi Y (2014) Structure and mechanism of a eukaryotic transmembrane ascorbate-dependent oxidoreductase. Proc Natl Acad Sci USA 111(5):1813–1818. CrossRefPubMedGoogle Scholar
  18. Mantzouridou F, Tsimidou MZ (2008) Lycopene formation in Blakeslea trispora. Chemical aspects of a bioprocess. Trends Food Sci Technol 19(7):363–371CrossRefGoogle Scholar
  19. Martin VJ, Pitera DJ, Withers ST, Newman JD, Keasling JD (2003) Engineering a mevalonate pathway in Escherichia coli for production of terpenoids. Nat Biotechnol 21(7):796–802. CrossRefPubMedGoogle Scholar
  20. Michael MR, Bausch J (2003) Summary of safety studies conducted with synthetic lycopene. Regul Toxicol Pharmacol 37(2):274–285CrossRefGoogle Scholar
  21. Nagao A, Yoshikawa T (2009) Absorption and function of dietary carotenoids. Forum Nutr 61:55CrossRefPubMedGoogle Scholar
  22. Rao AV, Agarwal S (2000) Role of antioxidant lycopene in cancer and heart disease. J Am Coll Nutr 19(5):563–569CrossRefPubMedGoogle Scholar
  23. Tao S, Miao L, Li Q, Dai G, Lu F, Tao L, Zhang X, Ma Y (2014) Production of lycopene by metabolically-engineered Escherichia coli. Biotechnol Lett 36(7):1515–1522CrossRefGoogle Scholar
  24. van Weeghel RP, Keck W, Robillard GT (1990) Regulated high-level expression of the mannitol permease of the phosphoenolpyruvate-dependent sugar phosphotransferase system in Escherichia coli. Proc Natl Acad Sci USA 87(7):2613–2617CrossRefPubMedGoogle Scholar
  25. Wang HH, Isaacs FJ, Carr PA, Sun ZZ, Xu G, Forest CR, Church GM (2009) Programming cells by multiplex genome engineering and accelerated evolution. Nature 460(7257):894–898CrossRefPubMedPubMedCentralGoogle Scholar
  26. Wu T, Ye L, Zhao D, Li S, Li Q, Zhang B, Bi C, Zhang X (2017) Membrane engineering—A novel strategy to enhance the production and accumulation of β-carotene in Escherichia coli. Metab Eng 43 Part A:85–91. CrossRefGoogle Scholar
  27. Yoon SH, Lee YM, Kim JE, Lee SH, Lee JH, Kim JY, Jung KH, Shin YC, Keasling JD, Kim SW (2006) Enhanced lycopene production in Escherichia coli engineered to synthesize isopentenyl diphosphate and dimethylallyl diphosphate from mevalonate. Biotechnol Bioeng 94(6):1025–1032CrossRefPubMedGoogle Scholar
  28. Yoon SH, Park HM, Kim JE, Lee SH, Choi MS, Kim JY, Oh DK, Keasling JD, Kim SW (2007) Increased beta-carotene production in recombinant Escherichia coli harboring an engineered isoprenoid precursor pathway with mevalonate addition. Biotechnol Prog 23(3):599–605. CrossRefPubMedGoogle Scholar
  29. Yoon SH, Lee SH, Das A, Ryu HK, Jang HJ, Kim JY, Oh DK, Keasling JD, Kim SW (2009) Combinatorial expression of bacterial whole mevalonate pathway for the production of beta-carotene in E. coli. J Biotechnol 140(3–4):218–226. CrossRefPubMedGoogle Scholar
  30. Yuan L, Rouviere P, Larossa R, Suh W (2006) Chromosomal promoter replacement of the isoprenoid pathway for enhancing carotenoid production in E. coli. Metab Eng 8(1):79–90. CrossRefPubMedGoogle Scholar
  31. Zhao D, Yuan S, Xiong B, Sun H, Ye L, Li J, Zhang X, Bi C (2016) Development of a fast and easy method for Escherichia coli genome editing with CRISPR/Cas9. Microb Cell Factories 15(1):205CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.College of Biological Sciences and TechnologyBeijing Forestry UniversityBeijingPeople’s Republic of China
  2. 2.Tianjin Institute of Industrial BiotechnologyChinese Academy of SciencesTianjinPeople’s Republic of China

Personalised recommendations