Skip to main content
SpringerLink
Log in

Screening, Isolation, and Identification of Zeaxanthin-Producing Bacteria

  • Protocol
  • First Online:
Microbial Carotenoids

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1852))

Abstract

Zeaxanthin is a yellow xanthophyll, dihydroxy-carotenoid, that is naturally found in some of the green, orange, and yellow vegetables and fruits and has a powerful antioxidant activity. Epidemiological evidences suggest that increasing the consumption of zeaxanthin in the diet is associated with a lower risk of age-related macular degeneration (ARMD) and cataracts, two of the leading causes of blindness in the world. Zeaxanthin is a promising nutraceutical/colorant with many applications in feed, food, and pharmaceutical industries. Currently, the commercial production of zeaxanthin is dependent on synthetic routes with limitation in production from biological sources. However, the biotechnological production of natural zeaxanthin is favored due to its safety, potential large-scale production and consumers’ preference for natural additives. In this chapter, we describe a rapid screening method based on 16S rRNA gene sequencing and effective HPLC with diode array detector/MS methods for the isolation and identification of zeaxanthin-producing bacteria and their carotenoid analysis.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 109.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 139.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 219.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Sommerburg O et al (1998) Fruits and vegetables that are sources for lutein and zeaxanthin: the macular pigment in human eyes. Br J Ophthalmol 82(8):907–910

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Nelis HJ, Deleenheer AP (1991) Microbial sources of carotenoid-pigments used in foods and feeds. J Appl Bacteriol 70(3):181–191

    Article  CAS  Google Scholar 

  3. Handelman GJ et al (1999) Antioxidant capacity of oat (Avena sativa L.) extracts. 1. Inhibition of low-density lipoprotein oxidation and oxygen radical absorbance capacity. J Agric Food Chem 47(12):4888–4893

    Article  CAS  PubMed  Google Scholar 

  4. Humphries JM, Khachik F (2003) Distribution of lutein, zeaxanthin, and related geometrical isomers in fruit, vegetables, wheat, and pasta products. J Agric Food Chem 51(5):1322–1327

    Article  CAS  PubMed  Google Scholar 

  5. Hadden WL et al (1999) Carotenoid composition of marigold (Tagetes erecta) flower extract used as nutritional supplement. J Agric Food Chem 47(10):4189–4194

    Article  CAS  PubMed  Google Scholar 

  6. Perry A, Rasmussen H, Johnson EJ (2009) Xanthophyll (lutein, zeaxanthin) content in fruits, vegetables and corn and egg products. J Food Compos Anal 22(1):9–15

    Article  CAS  Google Scholar 

  7. Khachik F et al (2002) Transformations of selected carotenoids in plasma, liver, and ocular tissues of humans and in nonprimate animal models. Invest Ophthalmol Vis Sci 43(11):3383–3392

    PubMed  Google Scholar 

  8. Snodderly DM (1995) Evidence for protection against age-related macular degeneration by carotenoids and antioxidant vitamins. Am J Clin Nutr 62(6):S1448–S1461

    Article  Google Scholar 

  9. Schweitzer D et al (2010) Suppelementation with lutein and zeaxanthin—a possible protection against age-related macular degeneration. Spektrum Augenheilkd 24(4):242–247

    Article  Google Scholar 

  10. Moeller SM et al (2008) Associations between age-related nuclear cataract and lutein and zeaxanthin in the diet and serum in the carotenoids in the age-related eye disease study, an ancillary study of the women's health initiative. Arch Ophthalmol 126(3):354–364

    Article  PubMed  PubMed Central  Google Scholar 

  11. Keri L et al (1997) The carotenoids β-carotene, canthaxanthin and zeaxanthin inhibit macrophage-mediated LDL oxidation. FEBS Lett 401:262–266

    Article  Google Scholar 

  12. Scripsema NK, Hu D-N, Rosen RB (2015) Lutein, zeaxanthin, and meso-zeaxanthin in the clinical management of eye disease. J Ophthalmol 865179:1–13

    Article  CAS  Google Scholar 

  13. Lindbergh CA et al (2017) Relationship of lutein and zeaxanthin levels to neurocognitive functioning: an fMRI study of older adults. J Int Neuropsychol Soc 23(1):11–22

    Article  PubMed  Google Scholar 

  14. Xu J et al (2017) Carotenoids and risk of fracture: a meta-analysis of observational studies. Oncotarget 8(2):2391

    Article  PubMed  Google Scholar 

  15. Liu Y et al (2017) Precise regulation of miR-210 is critical for the cellular homeostasis maintenance and transplantation efficacy enhancement of Mesenchymal stem cells in acute liver failure therapy. Cell Transplant 26(5):805–820

    Article  PubMed  PubMed Central  Google Scholar 

  16. Chen F et al (2017) Carotenoid intake and risk of non-Hodgkin lymphoma: a systematic review and dose-response meta-analysis of observational studies. Ann Hematol 96(6):957–965

    Article  CAS  PubMed  Google Scholar 

  17. Nishino H et al (2002) Carotenoids in cancer chemoprevention. Cancer Metastasis Rev 21(3–4):257–264

    Article  CAS  PubMed  Google Scholar 

  18. Nishino H et al (2000) Cancer prevention by natural carotenoids. Biofactors 13(1–4):89–94

    Article  CAS  PubMed  Google Scholar 

  19. Asker D, Beppu T, Ueda K (2007) Zeaxanthinibacter enoshimensis gen. nov., sp. nov., a novel zeaxanthin-producing marine bacterium of the family Flavobacteriaceae, isolated from seawater off Enoshima Island, Japan. Int J Syst Evol Microbiol 57(Pt 4):837–843

    Article  CAS  PubMed  Google Scholar 

  20. Asker D, Beppu T, Ueda K (2007) Mesoflavibacter zeaxanthinifaciens gen. nov., sp. nov., a novel zeaxanthin-producing marine bacterium of the family Flavobacteriaceae. Syst Appl Microbiol 30(4):291–296

    Article  CAS  PubMed  Google Scholar 

  21. Berry A et al (2003) Paracoccus zeaxanthinifaciens sp. nov., a zeaxanthin-producing bacterium. Int J Syst Evol Microbiol 53(Pt 1):231–238

    Article  CAS  PubMed  Google Scholar 

  22. Hoshino T, Ojima K, Setoguchi Y (2004) Production of zeaxanthin by recombinant Phaffia rhodozyma strain. PCT Int Appl US 20030923

    Google Scholar 

  23. Bhosale P, Bernstein PS (2005) Microbial xanthophylls. Appl Microbiol Biotechnol 68(4):445–455

    Article  CAS  PubMed  Google Scholar 

  24. Dufossé L (ed) (2009) Microbial and microalgal carotenoids as colourants and supplements. Birkhäuser Verlag, Basel

    Google Scholar 

  25. März U (2011) FOD025C—the global market for carotenoids. BCC Research, Wellesley, MA

    Google Scholar 

  26. Bosma TL, Dole JM, Maness NO (2003) Optimizing marigold (Tagetes erecta L.) petal and pigment yield. Crop Sci 43(6):2118–2124

    Article  Google Scholar 

  27. Stankovic I (2004) Zeaxanthin chemical and technical assessment (CTA). In: FAO (ed) 63rd JECFA

    Google Scholar 

  28. Gierhart DL (1994) Production of zeaxanthin and zeaxanthin-containing compositions. Applied Food Biotechnology, Inc. US 5,308,759

    Google Scholar 

  29. Chen F et al (2005) Isolation and purification of the bioactive carotenoid zeaxanthin from the microalga Microcystis aeruginosa by high-speed counter-current chromatography. J Chromatogr A 1064(2):183–186

    Article  CAS  PubMed  Google Scholar 

  30. Liao WL et al (1993) Pigmentation of cultured black tiger prawn by feeding with a Spirulina supplemented diet. Nippon Suisan Gakkaishi/Bull Jpn Soc Sci Fish 59:165–169

    Article  CAS  Google Scholar 

  31. Murata N et al (1981) Separation and characterization of thylakoid and cell envelope of the blue-green alga (cyanobacterium) Anacystis nidulans. Plant Cell Physiol 22(5):855–866

    CAS  Google Scholar 

  32. Withers NW et al (1978) Photosynthetic unit size, carotenoids, and chlorophyll-protein composition of Prochloron sp., a prokaryotic green alga. Proc Natl Acad Sci U S A 75(5):2301–2305

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Liao HH et al. (1995) Carotenoid producing culture using Nespongiococcum excentricum. US Patent 5,437,997

    Google Scholar 

  34. Jin ES, Feth B, Melis A (2003) A mutant of the green alga Dunaliella salina constitutively accumulates zeaxanthin under all growth conditions. Biotechnol Bioeng 81(1):115–124

    Article  CAS  PubMed  Google Scholar 

  35. Andrew Y, Britton G (1990) Photobleaching in the unicellular green alga Dunaliella parva 19/9. Photosynth Res 25:129–136

    Article  Google Scholar 

  36. Ben-Amotz A, Katz A, Avron M (1982) Accumulation of β-carotene in halotolerant algae: purification and characterization of β-carotene rich globules from Dunaliella bardawil (chlorophyceae). J Phycol 18:529–537

    Article  CAS  Google Scholar 

  37. Goodwin TW (1976) Distribution of carotenoids. In: Goodwin TW (ed) Chemistry and biochemistry of plant pigments. Academic Press Inc., New York

    Google Scholar 

  38. Palermo JA, Gros EG, Seldes AM (1991) Carotenoids from three red algae of the Corallinaceae. Phytochemistry 30(9):2983–2986

    Article  CAS  Google Scholar 

  39. Bhosale P et al (2004) Identification and characterization of a pi isoform of glutathione S-transferase (GSTP1) as a zeaxanthin-binding protein in the macula of the human eye. J Biol Chem 279(47):49447–49454

    Article  CAS  PubMed  Google Scholar 

  40. Lagarde D, Beuf L, Vermaas W (2000) Increased production of zeaxanthin and other pigments by application of genetic engineering techniques to Synechocystis sp. strain PCC 6803. Appl Environ Microbiol 66(1):64–72

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Hundle BS et al (1993) In vitro expression and activity of lycopene cyclase and beta-carotene hydroxylase from Erwinia herbicola. FEBS Lett 315(3):329–334

    Article  CAS  PubMed  Google Scholar 

  42. Joshi C, Singhal RS (2018) Zeaxanthin production by Paracoccus zeaxanthinifaciens ATCC 21588 in a lab-scale bubble column reactor: artificial intelligence modelling for determination of optimal operational parameters and energy requirements. Korean J Chem Eng 35(1):195–203

    Article  CAS  Google Scholar 

  43. Albrecht M, Misawa N, Sandmann G (1999) Metabolic engineering of the terpenoid biosynthetic pathway of Escherichia coli for production of the carotenoids beta-carotene and zeaxanthin. Biotechnol Lett 21(9):791–795

    Article  CAS  Google Scholar 

  44. Asker D, Beppu T, Ueda K (2007) Unique diversity of carotenoid-producing bacteria isolated from Misasa, a radioactive site in Japan. Appl Microbiol Biotechnol 77(2):383–392

    Article  CAS  PubMed  Google Scholar 

  45. Asker D, Beppu T, Ueda K (2008) Nubsella zeaxanthinifaciens gen. nov., sp. nov., a zeaxanthin-producing bacterium of the family Sphingobacteriaceae isolated from freshwater. Int J Syst Evol Microbiol 58(Pt 3):601–606

    Article  CAS  PubMed  Google Scholar 

  46. Kametani K, Matsumura T (1983) Determination of 238U, 234U, 226Ra and 228Ra in spring waters of sanin district. Radioisotopes 32(1):18–21

    Article  CAS  PubMed  Google Scholar 

  47. Asker D et al (2012) Isolation, characterization, and diversity of novel radiotolerant carotenoid-producing bacteria. In: Barredo J-L (ed) Microbial carotenoids from bacteria and microalgae: methods and protocols. Humana Press, New York, pp 21–60

    Chapter  Google Scholar 

  48. Asker D, Beppu T, Ueda K (2007) Sphingomonas jaspsi sp. nov., a novel carotenoid-producing bacterium isolated from Misasa, Tottori, Japan. Int J Syst Evol Microbiol 57(Pt 7):1435–1441

    Article  CAS  PubMed  Google Scholar 

  49. Sajilata MG, Singhal RS, Kamat MY (2008) The carotenoid pigment Zeaxanthin—a review. Compr Rev Food Sci Food Saf 7(1):29–49

    Article  CAS  Google Scholar 

  50. Jenkins CL et al (1979) The pigment of Pseudomonas paucimobilis is a carotenoid (nostoxanthin), rather than a brominated aryl-polyene (xanthomonadin). Curr Microbiol 3(1):1–4

    Article  CAS  Google Scholar 

  51. Rowe NJ et al (2000) Lipid composition and taxonomy of [Pseudomonas] echinoides: transfer to the genus Sphingomonas. Microbiology 146(Pt 11):3007–3012

    Article  CAS  PubMed  Google Scholar 

  52. Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4(4):406–425

    PubMed  CAS  Google Scholar 

  53. Asker D et al (2012) Novel zeaxanthin-producing bacteria isolated from a radioactive hot spring water. In: Barredo J-L (ed) Microbial carotenoids from bacteria and microalgae: methods and protocols. Humana Press, New York, pp 99–131

    Chapter  Google Scholar 

  54. Smibert RM, Krieg NR (1994) Phenotypic characterization. In: Gerhardt P, Murray RGE, Wood WA, Krieg NR (eds) Methods for general and molecular bacteriology. American Society for Microbiology, Washington, DC, pp 607–654

    Google Scholar 

  55. Beveridge TJ, Popkin TJ, Cole RM (1994) Electron microscopy. In P. Gerhardt (ed.), Methods for general molecular bacteriology. American Society for Microbiology, Washington, D.C., pp 42–71

    Google Scholar 

  56. Norris JR, Ribbons DW, Varma AK (1985) Methods in microbiology. Academic Press, London

    Google Scholar 

  57. Cowan ST, Steel KJ (1993) Manual for the identification of medical bacteria. Cambridge University Press, London

    Google Scholar 

  58. Collins MD (1994) Isoprenoid quinones. In: O’Donnell MGAG (ed) Chemical methods in prokaryotic systematics. Wiley, Chichester, pp 265–310

    Google Scholar 

  59. Kawahara K et al (1991) Chemical structure of glycosphingolipids isolated from Sphingomonas paucimobilis. FEBS Lett 292(1–2):107–110

    PubMed  CAS  Google Scholar 

  60. Takeuchi M, Hamana K, Hiraishi A (2001) Proposal of the genus Sphingomonas sensu stricto and three new genera, Sphingobium, Novosphingobium and Sphingopyxis, on the basis of phylogenetic and chemotaxonomic analyses. Int J Syst Evol Microbiol 51(Pt 4):1405–1417

    Article  CAS  PubMed  Google Scholar 

  61. Tindall BJ (1990) Lipid composition of Halobacterium lacusprofundi. FEMS Microbiol Lett 66:199–202

    Article  CAS  Google Scholar 

  62. Mesbah M, Whitman WB (1989) Measurement of deoxyguanosine/thymidine ratios in complex mixtures by high-performance liquid chromatography for determination of the mole percentage guanine + cytosine of DNA. J Chromatogr 479(2):297–306

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Food Science and Technology Department, Faculty of Agriculture, Alexandria University, Alexandria, Egypt

    Dalal Asker

  2. Department of Materials Science and Engineering, University of Toronto, Toronto, ON, Canada

    Dalal Asker & Tarek S. Awad

  3. Life Science Research Center, College of Bioresource Sciences, Nihon University, Fujisawa, Japan

    Teruhiko Beppu & Kenji Ueda

Authors
  1. Dalal Asker
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Tarek S. Awad
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Teruhiko Beppu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Kenji Ueda
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. Instituto de BiotecnologĂ­a de LeĂłn (INBIOTEC), Parque CientĂ­fico de LeĂłn, LeĂłn, Spain

    Carlos Barreiro

  2. Department of Biotechnology, Crystal Pharma, A Subsidiary of Albany, Molecular Research Inc. (AMRI), Parque TecnolĂłgico de LeĂłn, LeĂłn, Spain

    José-Luis Barredo

Rights and permissions

Reprints and permissions

Copyright information

© 2018 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Asker, D., Awad, T.S., Beppu, T., Ueda, K. (2018). Screening, Isolation, and Identification of Zeaxanthin-Producing Bacteria. In: Barreiro, C., Barredo, JL. (eds) Microbial Carotenoids. Methods in Molecular Biology, vol 1852. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8742-9_11

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-8742-9_11

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8741-2

  • Online ISBN: 978-1-4939-8742-9

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation