Skip to main content
SpringerLink
Log in

Characterization of a Deep-Sea Sediment Metagenomic Clone that Produces Water-Soluble Melanin in Escherichia coli

  • Original Article
  • Published:
Marine Biotechnology Aims and scope Submit manuscript

Abstract

To access to the microbial genetic resources of deep-sea sediment by a culture-independent approach, the sediment DNA was extracted and cloned into fosmid vector (pCC1FOS) generating a library of 39,600 clones with inserts of 24–45 kb. The clone fss6 producing red-brown pigment was isolated and characterized. The pigment was identified as melanin according to its physico-chemical characteristics. Subcloning and sequences analyses of fss6 demonstrated that one open reading frame (ORF2) was responsible for the pigment production. The deduced protein from ORF2 revealed significant amino acid similarity to the 4-hydroxyphenylpyruvate dioxygenase (HPPD) from deep-sea bacteria Idiomarina loihiensis. Further study demonstrated that the production of melanin was correlated with homogentistic acid (HGA). The p-hydroxyphenylpyruvate produced by the Escherichia coli host was converted to HGA through the oxidation reaction of introduced HPPD. The results demonstrate that expression of DNA extracted directly from the environment might generate applicable microbial gene products. The construction and analysis of the metagenomic library from deep-sea sediment contributed to our understanding for the reservoir of unexploited deep-sea microorganisms.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Brettar I, Christen R, Höfle MG (2003) Idiomarina baltica sp. nov., a marine bacterium with a high optimum growth temperature isolated from surface water of the central Baltic Sea. Int J Syst Evol Microbiol 53:407–413

    Article  PubMed  CAS  Google Scholar 

  • Choi DH, Cho BC (2005) Idiomarina seosinensis sp. nov., isolated from hypersaline water of a solar saltern in Korea. Int J Syst Evol Microbiol 55:379–383

    Article  PubMed  CAS  Google Scholar 

  • Daniel R (2005) The metagenomics of soil. Nat Rev Microbiol 3:470–478

    Article  PubMed  CAS  Google Scholar 

  • David C, Daro A, Szalai E, Atarhouch T, Mergeay M (1996) Formation of polymeric pigments in the presence of bacteria and comparison with chemical oxidative coupling-II. Catabolism of tyrosine and hydroxyphenylacetic acid by Alcaligenes eutrophus CH34 and mutants. Eur Polym J 32:669–679

    Article  CAS  Google Scholar 

  • Deming JW (1998) Deep ocean environmental biotechnology. Curr Opin Microbiol 9:283–287

    CAS  Google Scholar 

  • Donachie SP, Hou S, Gregory TS, Malahoff A, Alam M (2003) Idiomarina loihiensis sp. nov., a halophilic gamma-proteobacterium from the Lō’ihi submarine volcano, Hawai’i. Int J Syst Evol Microbiol 53:1873–1879

    Article  PubMed  CAS  Google Scholar 

  • Gibson LF, George AM (1998) Melanin and novel melanin precursors from Aeromonas media. FEMS Microbiol Lett 169:261–268

    Article  CAS  Google Scholar 

  • Gillespie DE, Brady SF, Bettermann AD, Cianciotto NP, Liles MR, Rondon MR, Clardy J, Goodman RM, Handelsman J (2002) Isolation of antibiotics turbomycin A and B from a metagenomic library of soil microbial DNA. Appl Environ Microbiol 68:4301–4306

    Article  PubMed  CAS  Google Scholar 

  • Hou S, Saw JH, Lee KS, Freitas TA, Belisle C, Kawarabayasi Y, Donachie SP, Pikina A, Galperin MY, Koonin EV, Makarova KS, Omelchenko MV, Sorokin A, Wolf YI, Li QX, Keum YS, Campbell S, Denery J, Aizawa S, Shibata S, Malahoff A, Alam M (2004) Genome sequence of the deep-sea γ-proteobacterium Idiomarina loihiensis reveals amino acid fermentation as a source of carbon and energy. Proc Natl Acad Sci U S A 101:18036–18041

    Article  PubMed  CAS  Google Scholar 

  • Ivanova EP, Romanenko LA, Chun J, Matte MH, Matte GR, Mikhailov VV, Svetashev VI, Huq A, Maugel T, Colwell RR (2000) Idiomarina gen. nov., comprising novel indigenous deep-sea bacteria from the Pacific Ocean, including descriptions of two species, Idiomarina abyssalis sp. nov. and Idiomarina zobellii sp. nov. Int J Syst Evol Microbiol 50:901–907

    PubMed  CAS  Google Scholar 

  • Keon J, Hargreaves J (1998) Isolation and heterologous expression of a gene encoding 4-hydroxyphenylpyruvate dioxygenase from the wheat leaf-spot pathogen, Mycosphaerella graminicola. FEMS Microbiol Lett 179:5862–5868

    Google Scholar 

  • Knox WE, Edwards SW (1955) Enzymes involved in conversion of tyrosine to acetoacetate. Methods Enzymol 2:287–300

    Article  CAS  Google Scholar 

  • Lagunas-Muñoz VH, Cabrera-Valladares N, Bolívar F, Gosset G, Martínez A (2006) Optimum melanin production using recombinant Escherichia coli. J Appl Microbiol 101:1002–1008

    Article  PubMed  Google Scholar 

  • Lai X, Cao L, Tan H, Fang S, Huang Y, Zhou S (2007) Fungal communities from methane hydrate-bearing deep-sea marine sediments in South China Sea. ISME J 1:756–762

    Article  PubMed  CAS  Google Scholar 

  • Lim HK, Chung EJ, Kim JC, Choi GJ, Jang KS, Chung YR, Cho KY, Lee SW (2005) Characterization of a forest soil metagenome clone that confers indirubin and indigo production on Escherichia coli. Appl Environ Microbiol 71:7768–7777

    Article  PubMed  CAS  Google Scholar 

  • Lindstedt S, Odelhög B (1987) 4-Hydroxyphenylpyruvate dioxygenase from human liver. Method Enzymol 142:139–142

    Article  CAS  Google Scholar 

  • Lindstedt S, Odelhög B, Rundgren M (1977) Purification and some properties of 4-hydroxyphenylpyruvate dioxygenase from Pseudomonas sp. P. J. 874. Biochemistry 16:3369–3377

    Article  PubMed  CAS  Google Scholar 

  • Lorenz P, Eck J (2005) Metagenomics and industrial applications. Nat Rev Microbiol 3:510–516

    Article  PubMed  CAS  Google Scholar 

  • MacNeil IA, Tiong CL, Minor C, August PR, Grossman TH, Loiacono KA, Lynch BA, Phillips T, Narula S, Sundaramoorthi R, Tyler A, Aldredge T, Long H, Gilman M, Holt D, Osburne MS (2001) Expression and isolation of antimicrobial small molecules from soil DNA libraries. J Mol Microbiol Biotechnol 3:301–308

    PubMed  CAS  Google Scholar 

  • Martínez-Cánovas MJ, Béjar V, Martínez-Checa F, Páez R, Quesada E (2004) Idiomarina fontislapidosi sp. nov. and Idiomarina ramblicola sp. nov., isolated from inland hypersaline habitats in Spain. Int J Syst Evol Microbiol 54:1793–1797

    Article  PubMed  Google Scholar 

  • Nichols MCA, Guezennec J, Bowman JP (2005) Bacterial exopolysaccharides from extreme marine environments with special consideration of the southern ocean, sea ice, and deep-sea hydrothermal vents: a review. Mar Biotechnol (NY) 7:253–271

    Article  CAS  Google Scholar 

  • Pittard AJ (1987) Biosynthesis of the aromatic amino acids. In: Neidhardt FC (ed) Escherichia coli and Salmonella typhimurium: cellular and molecular biology. ASM, Washington, DC, pp 368–394

    Google Scholar 

  • Riley PA (1997) Melanin. Int J Biochem Cell Biol 29:1235–1239

    Article  PubMed  CAS  Google Scholar 

  • Streit WR, Daniel R, Jaeger KE (2004) Prospecting for biocatalysts and drugs in the genomes of non-cultured microorganisms. Curr Opin Biotechnol 15:285–290

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgment

This work was supported by a grant from the National High Technology Research and Development Plan (863) of China (2007AA091904) and China Ocean Mineral Resources R&D Association (DYXM-115-02.2.14).

Author information

Authors and Affiliations

  1. State Key Laboratory for Biocontrol, Department of Biochemistry, College of Life Sciences, Sun Yat-sen University, Guangzhou, 510275, People’s Republic of China

    Yali Huang, Xiaocui He, Lixiang Cao, Zhirui Zeng, Jiong Zhang & Shining Zhou

  2. Shenzhen Academy of Metrology and Quality Inspection, Shenzhen, 518131, People’s Republic of China

    Xintian Lai

Authors
  1. Yali Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xintian Lai
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Xiaocui He
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Lixiang Cao
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Zhirui Zeng
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Jiong Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Shining Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Lixiang Cao or Shining Zhou.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Huang, Y., Lai, X., He, X. et al. Characterization of a Deep-Sea Sediment Metagenomic Clone that Produces Water-Soluble Melanin in Escherichia coli . Mar Biotechnol 11, 124–131 (2009). https://doi.org/10.1007/s10126-008-9128-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10126-008-9128-3

Keywords

Search

Navigation