Abstract
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.
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References
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. https://doi.org/10.1111/nph.13609
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):167
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):155–164
Bhataya A, Schmidt-Dannert C, Lee PC (2009) Metabolic engineering of Pichia pastoris X-33 for lycopene production. Process Biochem 44(10):1095–1102
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. https://doi.org/10.1128/aem.00115-10
Clinton SK (1998) Lycopene: chemistry, biology, and implications for human health and disease. Nutr Rev 56(2):35–51
Cronan JE Jr, Rock CO (2008) Biosynthesis of membrane lipids. EcoSal Plus. https://doi.org/10.1128/ecosalplus.3.6.4
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. https://doi.org/10.1074/jbc.M109.021618
Farmer WR, Liao JC (2000) Improving lycopene production in Escherichia coli by engineering metabolic control. Nat Biotechnol 18(5):533–537
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. https://doi.org/10.1083/jcb.200204144
Hillson NJ, Rosengarten RD, Keasling JD (2012) j5 DNA assembly design automation software. ACS Synth Biol 1(1):14–21. https://doi.org/10.1021/sb2000116
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):7
Jin YS, Stephanopoulos G (2007) Multi-dimensional gene target search for improving lycopene biosynthesis in Escherichia coli. Metab Eng 9(4):337–347. https://doi.org/10.1016/j.ymben.2007.03.003
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):636
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):489–497
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. https://doi.org/10.1007/s00253-011-3752-y
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. https://doi.org/10.1073/pnas.1323931111
Mantzouridou F, Tsimidou MZ (2008) Lycopene formation in Blakeslea trispora. Chemical aspects of a bioprocess. Trends Food Sci Technol 19(7):363–371
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. https://doi.org/10.1038/nbt833
Michael MR, Bausch J (2003) Summary of safety studies conducted with synthetic lycopene. Regul Toxicol Pharmacol 37(2):274–285
Nagao A, Yoshikawa T (2009) Absorption and function of dietary carotenoids. Forum Nutr 61:55
Rao AV, Agarwal S (2000) Role of antioxidant lycopene in cancer and heart disease. J Am Coll Nutr 19(5):563–569
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):1515–1522
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):2613–2617
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):894–898
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. https://doi.org/10.1016/j.ymben.2017.07.001
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):1025–1032
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. https://doi.org/10.1021/bp070012p
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. https://doi.org/10.1016/j.jbiotec.2009.01.008
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. https://doi.org/10.1016/j.ymben.2005.08.005
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):205
Acknowledgements
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).
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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.
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Wu, T., Ye, L., Zhao, D. et al. Engineering membrane morphology and manipulating synthesis for increased lycopene accumulation in Escherichia coli cell factories. 3 Biotech 8, 269 (2018). https://doi.org/10.1007/s13205-018-1298-8
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DOI: https://doi.org/10.1007/s13205-018-1298-8