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Biosynthesis and preparation of phycoerythrobilin in recombinant Escherichia coli

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Abstract

Phycoerythrobilin (PEB) is a special pigment in red algae and cyanobacteria that plays an important role in capturing light energy. As a natural non-toxic pigment with antioxidant properties, PEB is widely used in food and cosmetics industries. At present, the presence of other pigments in red algae and cyanobacteria, such as chlorophyll and phycocyanobilin, makes the preparation of PEB complicated and costly. In this study, the PEB synthesizing enzyme genes of hox1 from Synechocystis sp. PCC6803 and pebS from Nostoc sp. PCC 7120 were co-expressed in Escherichia coli to produce recombinant PEB. In addition, single-factor and response surface methods were used to enhance the yield of recombinant PEB. Under the conditions of lactose concentration at 4 mmol L−1, induction temperature at 27 °C, OD600 nm of 0.9, and induction time at 14 h, the concentration of PEB reached 20.37 mg L−1. In addition, a high-purity (purity > 70%) PEB was obtained through chloroform extraction, and the purified product was verified in UV spectrometry, high-performance liquid chromatography, and mass spectrometry.

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References

  • Apt KE, Collier JL, Grossman AR (1995) Evolution of the phycobiliproteins. J Mol Biol 248:79–96

    Article  CAS  Google Scholar 

  • Aras M, Hartmann V, Hartmann J, Nowaczyk MM, Frankenberg-Dinkel N (2020) Proximity channeling during cyanobacterial phycoerythrobilin synthesis. FEBS J 287:284–194

    Article  CAS  Google Scholar 

  • Camara-Artigas A, Bacarizo J, Andujar-Sanchez M, Ortiz-Salmeron E, Mesa-Valle C, Cuadri C, Martin-Garcia JM, Martinez-Rodriguez S, Mazzuca-Sobczuk T, Ibanez MJ, others (2012) pH-dependent structural conformations of B-phycoerythrin from Porphyridium cruentum. FEBS J 279:3680–3691

    Article  CAS  Google Scholar 

  • Chapman DJ, Cole WJ, Siegelman HW (1967) Structure of phycoerythrobilin. J Am Chem Soc 89:5976–5977

    Article  CAS  Google Scholar 

  • Dammeyer T, Frankenberg-Dinkel N (2006) Insights into phycoerythrobilin biosynthesis point toward metabolic channeling. J Biol Chem 281:27081–27089

    Article  CAS  Google Scholar 

  • Dammeyer T, Bagby SC, Sullivan MB, Chisholm SW, Frankenberg-Dinkel N (2008) Efficient phage-mediated pigment biosynthesis in oceanic cyanobacteria. Curr Biol 18:442–448

    Article  CAS  Google Scholar 

  • Davis SJ, Kurepa J, Vierstra RD (1999) The Arabidopsis thaliana HY1 locus, required for phytochrome-chromophore biosynthesis, encodes a protein related to heme oxygenases. Proc Nat Acad Sci USA 96:6541–6546

    Article  CAS  Google Scholar 

  • Eriksen NT (2008) Production of phycocyanin—a pigment with applications in biology, biotechnology, foods and medicine. Appl Microbiol Biotechnol 80:1–14

    Article  CAS  Google Scholar 

  • Frankenberg N, Mukougawa K, Kohchi T, Lagarias JC (2001) Functional genomic analysis of the HY2 family of ferredoxin-dependent bilin reductases from oxygenic photosynthetic organisms. Plant Cell 13:965–978

    Article  CAS  Google Scholar 

  • Gaigalas A, Gallagher T, Cole KD, Singh T, Wang LL, Zhang YZ (2006) A multistate model for the fluorescence response of R-phycoerythrin. Photochem Photobiol 82:635–644

    Article  CAS  Google Scholar 

  • Gambetta GA, Lagarias JC (2001) Genetic engineering of phytochrome biosynthesis in bacteria. Proc Nat Acad Sci U S A 98:10566–10571

    Article  CAS  Google Scholar 

  • Ge B, Li Y, Sun H, Zhang S, Hu P, Qin S, Huang F (2013) Combinational biosynthesis of phycocyanobilin using genetically-engineered Escherichia coli. Biotechnol Lett 35:689–693

    Article  CAS  Google Scholar 

  • Glazer AN, Hixson CS (1977) Subunit structure and chromophore composition of rhodophytan phycoerythrins. Porphyridium cruentum B-phycoerythrin and b-phycoerythrin. J Biol Chem 252:32–42

  • Hou YN, Ding WL, Hu JL, Jiang XX, Tan ZZ, Zhao KH (2019) Very bright phycoerythrobilin chromophore for fluorescence biolabeling. Chembiochem 20:2777–2783

    Article  CAS  Google Scholar 

  • Jiang T, Zhang JP, Liang DC (1999) Structure and function of chromophores in R-phycoerythrin at 1.9 angstrom resolution. Proteins 34:224–231

    Article  CAS  Google Scholar 

  • Kehoe DM, Grossman AR (1994) Complementary chromatic adaptation: photoperception to gene regulation. Semin Cell Biol 5:303–313

    Article  CAS  Google Scholar 

  • Lee D, Nishizawa M, Shimizu Y, Saeki H (2017) Anti-inflammatory effects of dulse (Palmaria palmata) resulting from the simultaneous water-extraction of phycobiliproteins and chlorophyll a. Food Res Int 100:514–521

    Article  CAS  Google Scholar 

  • Li WJ, Pu Y, Ge BS, Wang YC, Yu DY, Qin S (2019) Dye-sensitized solar cells based on natural and artificial phycobiliproteins to capture low light underwater. Int J Hydrogen Energ 44:1182–1191

    Article  CAS  Google Scholar 

  • Liu Y, de Montellano PRO (2000) Reaction intermediates and single turnover rate constants for the oxidation of heme by human heme oxygenase-1. J Biol Chem 275:5297–5307

    Article  CAS  Google Scholar 

  • Liu S, Chen H, Qin S, Zhang W, Guan X, Lu Y (2009) Highly soluble and stable recombinant holo-phycocyanin alpha subunit expressed in Escherichia coli. Biochem Eng J 48:58–64

    Article  CAS  Google Scholar 

  • Mittal R, Sharma R, Raghavarao KSMS (2019) Aqueous two-phase extraction of R-Phycoerythrin from marine macro-algae, Gelidium pusillum. Bioresour Technol 280:277–286

    Article  CAS  Google Scholar 

  • Montellano PROD (2000) The mechanism of heme oxygenase. Curr Opin Chem Biol 4:221–227

    Article  CAS  Google Scholar 

  • Muramoto T, Kohchi T, Yokota A, Hwang IH, Goodman HM (1999) The Arabidopsis photomorphogenic mutant hy1 is deficient in phytochrome chromophore biosynthesis as a result of a mutation in a plastid heme oxygenase. Plant Cell 11:335–347

    Article  CAS  Google Scholar 

  • Muramoto T, Tsurui N, Terry MJ, Yokota A, Kohchi T (2002) Expression and biochemical properties of a ferredoxin-dependent heme oxygenase required for phytochrome chromophore synthesis. Plant Physiol 130:1958–1966

    Article  CAS  Google Scholar 

  • Ocarra P, Murphy RF, Killilea SD (1980) The native forms of the phycobilin chromophores of algal biliproteins — A clarification. Biochem J 187:303–309

    Article  CAS  Google Scholar 

  • Oh JH, Kim EY, Nam TJ (2018) Phycoerythrin peptide from Pyropia yezoensis alleviates endoplasmic reticulum stress caused by perfluorooctane sulfonate-induced calcium dysregulation. Mar Drugs 16:44–58

    Article  Google Scholar 

  • Parmar A, Singh NK, Kaushal A, Sonawala S, Madamwar D (2011) Purification, characterization and comparison of phycoerythrins from three different marine cyanobacterial cultures. Bioresour Technol 102:1795–1802

    Article  CAS  Google Scholar 

  • Rockwell NC, Martin SS, Lagarias JC (2016) Identification of cyanobacteriochromes detecting far-red light. Biochemistry 33:3907–3919

    Article  Google Scholar 

  • Sakai S, Komura Y, Nishimura Y, Sugawara T, Hirata T (2011) Inhibition of mast cell degranulation by phycoerythrin and its pigment moiety phycoerythrobilin, prepared from Porphyra yezoensis. Food Sci Technol Res 17:171–177

    Article  CAS  Google Scholar 

  • Sekar S, Chandramohan M (2008) Phycobiliproteins as a commodity: trends in applied research, patents and commercialization. J Appl Phycol 20:113–136

    Article  Google Scholar 

  • Stiefelmaier J, Ledermann B, Sorg M, Banek A, Geib D, Ulber R, Frankenberg-Dinkel N (2018) Pink bacteria-Production of the pink chromophore phycoerythrobilin with Escherichia coli. J Biotechnol 274:47–53

    Article  CAS  Google Scholar 

  • Terry MJ, Linley PJ, Kohchi T (2002) Making light of it: the role of plant haem oxygenases in phytochrome chromophore synthesis. Biochem Soc Trans 30:604–609

    Article  Google Scholar 

  • Wu J, Chen HX, Zhao J, Jiang P (2017) Fusion proteins of streptavidin and allophycocyanin alpha subunit for immunofluorescence assay. Biochem Eng J 125:97–103

    Article  CAS  Google Scholar 

  • Yabuta Y, Fujimura H, Kwak CS, Enomoto T, Watanabe F (2010) Antioxidant activity of the phycoerythrobilin compound formed from a dried Korean Purple Laver (Porphyra sp.) during in vitro digestion. Food Sci Technol Res 16:347–351

    Article  CAS  Google Scholar 

Download references

Funding

This work was supported by the National Natural Science Foundation of China (Nos. 41906109, 42061134020, and 41411130202) and the Key Research and Development Program of Yantai City, Shandong, China (Nos. 2019XDHZ101 and 2020MSGY084).

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Correspondence to Song Qin or Jian Lin.

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Li, W., Ma, C., Ge, B. et al. Biosynthesis and preparation of phycoerythrobilin in recombinant Escherichia coli. J Appl Phycol 33, 1673–1683 (2021). https://doi.org/10.1007/s10811-021-02408-0

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  • DOI: https://doi.org/10.1007/s10811-021-02408-0

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