Skip to main content

Engineering E. coli for caffeic acid biosynthesis from renewable sugars

Applied Microbiology and Biotechnology volume 97, pages 3333–3341 (2013)Cite this article

Abstract

Caffeic acid is a valuable aromatic compound that possesses many important pharmacological activities. In structure, caffeic acid belongs to the hydroxycinnamic acid family and can be biosynthesized from the aromatic amino acid tyrosine. In the present paper, the caffeic acid biosynthesis pathway was reconstituted in engineered Escherichia coli to produce caffeic acid from simple biomass sugar glucose and xylose. Different engineering approaches were utilized to optimize the production. Specifically, two parallel biosynthesis routes leading from tyrosine to caffeic acid were studied. The copy number of the intermediate biosynthesis genes was varied to find appropriate gene doses for caffeic acid biosynthesis. Three different media, including a MOPS medium, a synthetic medium, and a rich medium, were also examined to improve the production. The highest specific caffeic acid production achieved was 38 mg/L/OD. Lastly, cultivation of engineered E. coli in a bioreactor resulted in a production of 106 mg/L caffeic acid after 4 days.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

43,34 €

Price includes VAT (Finland)
Tax calculation will be finalised during checkout.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

References

  • Berner M, Krug D, Bihlmaier C, Vente A, Müller R, Bechthold A (2006) Genes and enzymes involved in caffeic acid biosynthesis in the actinomycete Saccharothrix espanaensis. J Bacteriol 188(7):2666–2673

    Article  CAS  Google Scholar 

  • Celik S, Erdogan S, Tuzcu M (2009) Caffeic acid phenethyl ester (CAPE) exhibits significant potential as an antidiabetic and liver-protective agent in streptozotocin-induced diabetic rats. Pharmacol Res 60(4):270–276

    Article  CAS  Google Scholar 

  • Cheng JT, Liu IM, Tzeng TF, Chen WC, Hayakawa S, Yamamoto T (2003) Release of beta-endorphin by caffeic acid to lower plasma glucose in streptozotocin-induced diabetic rats. Horm Metab Res 35(4):251–258

    Article  CAS  Google Scholar 

  • Choi O, Wu CZ, Kang SY, Ahn JS, Uhm TB, Hong YS (2011) Biosynthesis of plant-specific phenylpropanoids by construction of an artificial biosynthetic pathway in Escherichia coli. J Ind Microbiol Biotechnol 38(10):1657–1665

    Article  CAS  Google Scholar 

  • Chuu CP, Lin HP, Ciaccio MF, Kokontis JM, Hause RJ Jr, Hiipakka RA, Liao S, Jones RB (2012) Caffeic acid phenethyl ester suppresses the proliferation of human prostate cancer cells through inhibition of p70S6K and Akt signaling networks. Cancer Prev Res (Phila) 5(5):788–797

    Article  CAS  Google Scholar 

  • Gosset G (2009) Production of aromatic compounds in bacteria. Curr Opin Biotechnol 20(6):651–658

    Article  CAS  Google Scholar 

  • Gross GG, Zenk MH (1974) Isolation and properties of hydroxycinnamate:CoA ligase from lignifying tissue of Forsythia. Eur J Biochem 42(2):453–459

    Article  CAS  Google Scholar 

  • Grunberger D, Banerjee R, Eisinger K, Oltz EM, Efros L, Caldwell M, Estevez V, Nakanishi K (1988) Preferential cytotoxicity on tumor cells by caffeic acid phenethyl ester isolated from propolis. Experientia 44(3):230–232

    Article  CAS  Google Scholar 

  • Hudson EA, Dinh PA, Kokubun T, Simmonds MS, Gescher A (2000) Characterization of potentially chemopreventive phenols in extracts of brown rice that inhibit the growth of human breast and colon cancer cells. Cancer Epidemiol Biomark Prev 9(11):1163–1170

    CAS  Google Scholar 

  • Ikeda K, Tsujimoto K, Uozaki M, Nishide M, Suzuki Y, Koyama AH, Yamasaki H (2011) Inhibition of multiplication of herpes simplex virus by caffeic acid. Int J Mol Med 28(4):595–598

    CAS  Google Scholar 

  • Juminaga D, Baidoo EE, Redding-Johanson AM, Batth TS, Burd H, Mukhopadhyay A, Petzold CJ, Keasling JD (2012) Modular engineering of l-tyrosine production in Escherichia coli. Appl Environ Microbiol 78(1):89–98

    Article  CAS  Google Scholar 

  • Jung UJ, Lee MK, Park YB, Jeon SM, Choi MS (2006) Antihyperglycemic and antioxidant properties of caffeic acid in db/db mice. J Pharmacol Exp Ther 318(2):476–483

    Article  CAS  Google Scholar 

  • Kneusel RE, Matern U, Nicolay K (1989) Formation of trans-caffeoyl-CoA from trans-4-coumaroyl-CoA by Zn2+-dependent enzymes in cultured plant cells and its activation by an elicitor-induced pH shift. Arch Biochem Biophys 269(2):455–462

    Article  CAS  Google Scholar 

  • Leonardis AD, Macciola V, Domenico ND (2005) A first pilot study to produce a food antioxidant from sunflower seed shells (Helianthus annuus). Eur J Lipid Sci Technol 107(4):220–227

    Article  Google Scholar 

  • Lin Y, Yan Y (2012) Biosynthesis of caffeic acid in Escherichia coli using its endogenous hydroxylase complex. Microb Cell Fact 11:42

    Article  Google Scholar 

  • Lutke-Eversloh T, Stephanopoulos G (2008) Combinatorial pathway analysis for improved l-tyrosine production in Escherichia coli: identification of enzymatic bottlenecks by systematic gene overexpression. Metab Eng 10(2):69–77

    Article  Google Scholar 

  • Maurya DK, Devasagayam TP (2010) Antioxidant and prooxidant nature of hydroxycinnamic acid derivatives ferulic and caffeic acids. Food Chem Toxicol 48(12):3369–3373

    Article  CAS  Google Scholar 

  • McKenna R, Nielsen DR (2011) Styrene biosynthesis from glucose by engineered E. coli. Metab Eng 13(5):544–554

    Article  CAS  Google Scholar 

  • Mori H, Iwahashi H (2009) Antioxidant activity of caffeic acid through a novel mechanism under UVA irradiation. J Clin Biochem Nutr 45:49–55

    Article  CAS  Google Scholar 

  • Morton LW, Croft KD, Puddey IB, Byrne L (2000) Phenolic acids protect low density lipoproteins from peroxynitrite-mediated modification in vitro. Redox Rep 5(2–3):124–125

    Article  CAS  Google Scholar 

  • Nakagawa A, Minami H, Kim JS, Koyanagi T, Katayama T, Sato F, Kumagai H (2011) A bacterial platform for fermentative production of plant alkaloids. Nat Commun 2:326

    Article  Google Scholar 

  • Nambudiri AMD, Bhat JV (1972) Conversion of p-coumarate into caffeate by Streptomyces nigrifaciens. Purification and properties of the hydroxylating enzyme. Biochem J 130(2):425–433

    CAS  Google Scholar 

  • Natarajan K, Singh S, Burke TR Jr, Grunberger D, Aggarwal BB (1996) Caffeic acid phenethyl ester is a potent and specific inhibitor of activation of nuclear transcription factor NF-KB. Proc Natl Acad Sci USA 93(17):9090–9095

    Article  CAS  Google Scholar 

  • Qi WW, Vannelli T, Breinig S, Ben-Bassat A, Gatenby AA, Haynie SL, Sariaslani FS (2007) Functional expression of prokaryotic and eukaryotic genes in Escherichia coli for conversion of glucose to p-hydroxystyrene. Metab Eng 9(3):268–276

    Article  CAS  Google Scholar 

  • Rajendra-Prasad N, Karthikeyan A, Karthikeyan S, Reddy BV (2011) Inhibitory effect of caffeic acid on cancer cell proliferation by oxidative mechanism in human HT-1080 fibrosarcoma cell line. Mol Cell Biochem 349(1–2):11–19

    Article  CAS  Google Scholar 

  • Ramirez-AhumadaMdel C, Timmermann BN, Gang DR (2006) Biosynthesis of curcuminoids and gingerols in turmeric (Curcuma longa) and ginger (Zingiber officinale): identification of curcuminoid synthase and hydroxycinnamoyl-CoA thioesterases. Phytochemistry 67(18):2017–2029

    Article  Google Scholar 

  • Rosler J, Krekel F, Amrhein N, Schmid J (1997) Maize phenylalanine ammonia-lyase has tyrosine ammonia-lyase activity. Plant Physiol 113(1):175–179

    Article  CAS  Google Scholar 

  • Santos CN, Stephanopoulos G (2008) Melanin-based high-throughput screen for l-tyrosine production in Escherichia coli. Appl Environ Microbiol 74(4):1190–1197

    Article  CAS  Google Scholar 

  • Santos CN, Koffas M, Stephanopoulos G (2011) Optimization of a heterologous pathway for the production of flavonoids from glucose. Metab Eng 13(4):392–400

    Article  CAS  Google Scholar 

  • Santos CN, Xiao W, Stephanopoulos G (2012) Rational, combinatorial, and genomic approaches for engineering l-tyrosine production in Escherichia coli. Proc Natl Acad Sci USA 109(34):13538–13543

    Article  CAS  Google Scholar 

  • Satoh Y, Tajima K, Munekata M, Keasling JD, Lee TS (2012) Engineering of a tyrosol-producing pathway, utilizing simple sugar and the central metabolic tyrosine, in Escherichia coli. J Agric Food Chem 60(4):979–984

    Article  CAS  Google Scholar 

  • Song F, Zhuang Z, Finci L, Dunaway-Mariano D, Kniewel R, Buglino JA, Solorzano V, Wu J, Lima CD (2006) Structure, function and mechanism of the phenylacetate pathway hotdog-fold thioesterase PAAI. J Biol Chem 281(16):11028–11038

    Article  CAS  Google Scholar 

  • Sud'ina GF, Mirzoeva OK, Pushkareva MA, Korshunova GA, Sumbatyan NV, Varfolomeev SD (1993) Caffeic acid phenethyl ester as a lipoxygenase inhibitor with antioxidant properties. FEBS Lett 329(1–2):21–24

    Article  Google Scholar 

  • Takeda H, Tsuji M, Inazu M, Egashira T, Matsumiya T (2002) Rosmarinic acid and caffeic acid produce antidepressive-like effect in the forced swimming test in mice. Eur J Pharmacol 449(3):261–267

    Article  CAS  Google Scholar 

  • Verhoef S, Ruijssenaars HJ, de Bont JAM, Wery J (2007) Bioproduction of p-hydroxybenzoate from renewable feedstock by solvent-tolerant Pseudomonas putida S12. J Biotechnol 132:49–56

    Article  CAS  Google Scholar 

  • Wang J, Lu D, Zhao H, Ling X, Jiang B, Ouyang P (2009) Application of response surface methodology optimization for the production of caffeic acid from tobacco waste. Afr J Biotechnol 8(8):1416–1424

    CAS  Google Scholar 

  • Ye JC, Hsiao MW, Hsieh CH, Wu WC, Hung YC, Chang WC (2010) Analysis of caffeic acid extraction from Ocimum gratissimum Linn. by high performance liquid chromatography and its effects on a cervical cancer cell line. Taiwan J Obstet Gynecol 49(3):266–271

    Article  Google Scholar 

  • Zhang H, Wang Y, Pfeifer BA (2008) Bacterial hosts for natural product production. Mol Pharm 5(2):212–225

    Article  CAS  Google Scholar 

  • Zhuang Z, Song F, Zhao H, Li L, Cao J, Eisenstein E, Herzberg O, Dunaway-Mariano D (2008) Divergence of function in the hot dog fold enzyme superfamily: the bacterial thioesterase YciA. Biochemistry 47(9):2789–2796

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This study was supported by ARPA-E (DE-AR0000059) provided by US Department of Energy and the Singapore MIT Alliance.

Author information

Affiliations

  1. Department of Chemical Engineering, Massachusetts Institute of Technology, 77 Massachusetts Ave, Room 56-469, Cambridge, MA, 02139, USA

    Haoran Zhang & Gregory Stephanopoulos

Authors
  1. Haoran Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Gregory Stephanopoulos
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Gregory Stephanopoulos.

Electronic supplementary material

Below is the link to the electronic supplementary material.

ESM 1

DOCX 116 kb

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Zhang, H., Stephanopoulos, G. Engineering E. coli for caffeic acid biosynthesis from renewable sugars. Appl Microbiol Biotechnol 97, 3333–3341 (2013). https://doi.org/10.1007/s00253-012-4544-8

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00253-012-4544-8

Keywords

  • Escherichia coli
  • Caffeic acid
  • Tyrosine
  • Heterologous biosynthesis