Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Discovery of “heat shock metabolites” produced by thermotolerant actinomycetes in high-temperature culture

Abstract

In actinomycetes, many secondary metabolite biosynthetic genes are not expressed under typical laboratory culture conditions and various efforts have been made to activate these dormant genes. In this study, we focused on high-temperature culture. First, we examined the thermotolerance of 3160 actinomycete strains from our laboratory culture collection and selected 57 thermotolerant actinomycetes that grew well at 45 °C. These 57 thermotolerant actinomycetes were cultured for 5 days in liquid medium at both 30 °C and 45 °C. Culture broths were extracted with 1-butanol, and each extract was subjected to LC/MS analysis. The metabolic profiles of each strain were compared between the 30 °C and 45 °C cultures. We found that almost half of these thermotolerant actinomycetes produced secondary metabolites that were detected only in the 45 °C culture. This result suggests that high-temperature culture induces the production of dormant secondary metabolites. These compounds were named “heat shock metabolites (HSMs).” To examine HSM production in more detail, 18 strains were selected at random from the initial 57 strains and cultivated in six different media at 30 °C and 45 °C; as before, metabolic profiles of each strain in each medium were compared between the 30 °C and 45 °C cultures. From this analysis, we found a total of 131 HSMs. We identified several angucycline-related compounds as HSMs from two thermotolerant Streptomyces species. Furthermore, we discovered a new compound, murecholamide, as an HSM from thermotolerant Streptomyces sp. AY2. We propose that high-temperature culture of actinomycetes is a convenient method for activating dormant secondary metabolite biosynthetic genes.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

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

References

  1. 1.

    Dictionary of natural products on DVD, version 23.1. London, UK: Chapman & Hall/CRC;2014.

  2. 2.

    Ohnishi Y, et al. Genome sequence of the streptomycin-producing microorganism Streptomyces griseus IFO 13350. J Bacteriol. 2008;190:4050–60.

    CAS  Article  Google Scholar 

  3. 3.

    Bentley SD, et al. Complete genome sequence of the model actinomycete Streptomyces coelicolor A3(2). Nature. 2002;417:141–7.

    Article  Google Scholar 

  4. 4.

    Ikeda H, Kazuo SY, Omura S. Genome mining of the Streptomyces avermitilis genome and development of genome-minimized hosts for heterologous expression of biosynthetic gene clusters. J Ind Microbiol Biotechnol. 2014;41:233–50.

    CAS  Article  Google Scholar 

  5. 5.

    van Keulen G, Dyson PJ. Production of specialized metabolites by Streptomyces coelicolor A3(2). Adv Appl Microbiol. 2014;89:217–66.

    Article  Google Scholar 

  6. 6.

    Katz L, Baltz RH. Natural product discovery: past, present, and future. J Ind Microbiol Biotechnol. 2016;43:155–76.

    CAS  Article  Google Scholar 

  7. 7.

    Ochi K, et al. Ribosome engineering and secondary metabolite production. Adv Appl Microbiol. 2004;56:155–84.

    CAS  Article  Google Scholar 

  8. 8.

    Onaka H. Novel antibiotic screening methods to awaken silent or cryptic secondary metabolic pathways in actinomycetes. J Antibiot. 2017;70:865–70.

    CAS  Article  Google Scholar 

  9. 9.

    Onaka H, Mori Y, Igarashi Y, Furumai T. Mycolic acid-containing bacteria induce natural-product biosynthesis in Streptomyces species. Appl Environ Microbiol. 2011;77:400–6.

    CAS  Article  Google Scholar 

  10. 10.

    Asamizu S, Ozaki T, Teramoto K, Satoh K, Onaka H. Killing of mycolic acid-containing bacteria aborted induction of antibiotic production by Streptomyces in combined-culture. PLoS ONE. 2015;10:e0142372.

    Article  Google Scholar 

  11. 11.

    Keskar SS, Srinivasan MC, Deshpande VV. Chemical modification of a xylanase from a thermotolerant Streptomyces. Evidence for essential tryptophan and cysteine residues at the active site. Biochem J. 1989;261:49–55.

    CAS  Article  Google Scholar 

  12. 12.

    Doull JL, Ayer SW, Singh AK, Thibault P. Production of a novel polyketide antibiotic, jadomycin B, by Streptomyces venezuelae following heat shock. J Antibiot. 1993;46:869–71.

    CAS  Article  Google Scholar 

  13. 13.

    Wei ZH, Wu H, Bai L, Deng Z, Zhong JJ. Temperature shift-induced reactive oxygen species enhanced validamycin A production in fermentation of Streptomyces hygroscopicus 5008. Bioprocess Biosyst Eng. 2012;35:1309–16.

    CAS  Article  Google Scholar 

  14. 14.

    Akiyama H, Oku N, Harunari E, Panbangred W, Igarashi Y. Complete NMR assignment and absolute configuration of k4610422, a norditerpenoid inhibitor of testosterone-5α-reductase originally from Streptosporangium: rediscovery from a thermophilic Actinomadura. J Antibiot. 2019. https://doi.org/10.1038/s41429-019-0231-7.

    Article  PubMed  Google Scholar 

  15. 15.

    Teta R, et al. Thermoactinoamide A, an antibiotic lipophilic cyclopeptide from the Icelandic thermophilic bacterium Thermoactinomyces vulgaris. J Nat Prod. 2017;80:2530–5.

    CAS  Article  Google Scholar 

  16. 16.

    Hwang BK, Lim SW, Kim BS, Lee JY, Moon SS. Isolation and in vivo and in vitro antifungal activity of phenylacetic acid and sodium phenylacetate from Streptomyces humidus. Appl Environ Chem. 2001;67:3739–45.

    CAS  Article  Google Scholar 

  17. 17.

    Kern DL, Schaumberg JP, Hokanson GC, French JC. PD 116,779, a new antitumor antibiotic of the benz[a]anthraquinone class. J Antibiot. 1986;39:469–70.

    CAS  Article  Google Scholar 

  18. 18.

    Kesenheimer C, Kalogerakis A, Meissner A, Groth U. The cobalt way to angucyclinones: asymmetric total synthesis of the antibiotics (+)-rubiginone B2, (−)-tetrangomycin, and (−)-8-O-methyltetrangomycin. Chemistry. 2010;16:8805–21.

    CAS  Article  Google Scholar 

  19. 19.

    Yamashita N, Harada T, Shin-ya K, Seto H. 6-Hydroxytetrangulol, a new CPP32 protease inducer produced by Streptomyces sp. J Antibiot. 1998;51:79–81.

    CAS  Article  Google Scholar 

  20. 20.

    Kallio P, Liu Z, Mäntsälä P, Niemi J, Metsä-Ketelä M. Sequential action of two flavoenzymes, PgaE and PgaM, in angucycline biosynthesis: chemoenzymatic synthesis of gaudimycin C. Chem Biol. 2008;15:157–66.

    CAS  Article  Google Scholar 

  21. 21.

    Palmu K, Ishida K, Mäntsälä P, Hertweck C, Metsä-Ketelä M. Artificial reconstruction of two cryptic angucycline antibiotic biosynthetic pathways. Chembiochem. 2007;8:1577–84.

    CAS  Article  Google Scholar 

  22. 22.

    Rohr J, et al. Metabolic products of microorganisms. 249. Tetracenomycins B3 and D3, key intermediates of the elloramycin and tetracenomycin C biosynthesis. J Antibiot. 1988;41:1066–73.

    CAS  Article  Google Scholar 

  23. 23.

    Gorajana A, et al. Resistoflavine, cytotoxic compound from a marine actinomycete, Streptomyces chibaensis AUBN1/7. Microbiol Res. 2007;162:322–7.

    CAS  Article  Google Scholar 

  24. 24.

    Kock I, Maskey RP, Biabani MA, Helmke E, Laatsch H. 1-Hydroxy-1-norresistomycin and resistoflavin methyl ether: new antibiotics from marine-derived streptomycetes. J Antibiot. 2005;58:530–4.

    CAS  Article  Google Scholar 

  25. 25.

    Wang W, et al. Angucyclines as signals modulate the behaviors of Streptomyces coelicolor. Proc Natl Acad Sci USA. 2014;111:5688–93.

    CAS  Article  Google Scholar 

  26. 26.

    Barona-Gómez F, Wong U, Giannakopulos AE, Derrick PJ, Challis GL. Identification of a cluster of genes that directs desferrioxamine biosynthesis in Streptomyces coelicolor M145. J Am Chem Soc. 2004;126:16282–3.

    Article  Google Scholar 

  27. 27.

    Lee HS, et al. Cyclic peptides of the nocardamine class from a marine-derived bacterium of the genus Streptomyces. J Nat Prod. 2005;68:623–5.

    CAS  Article  Google Scholar 

  28. 28.

    Tokyo Chemical Industry Co., Ltd., Japan. https://www.tcichemicals.com/ja/jp/index.html (Accessed on Sep 29, 2019).

  29. 29.

    Drzyzga O, Fernández de las Heras L, Morales V, Navarro Llorens JM, Perera J. Cholesterol degradation by Gordonia cholesterolivorans. Appl Environ Microbiol. 2011;77:4802–10.

    CAS  Article  Google Scholar 

  30. 30.

    Nakae K, et al. Migrastatin, a new inhibitor of tumor cell migration from Streptomyces sp. MK929-43F1. Taxonomy, fermentation, isolation and biological activities. J Antibiot. 2000;53:1130–6.

    CAS  Article  Google Scholar 

Download references

Acknowledgements

This work was supported by a Grant-in-Aid for Scientific Research in Innovative Areas (Grant JP17H06401 to MI) from The Ministry of Education, Culture, Sports, Science and Technology (MEXT).

Author information

Affiliations

Authors

Corresponding authors

Correspondence to Yasuo Ohnishi or Masaya Imoto.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Saito, S., Kato, W., Ikeda, H. et al. Discovery of “heat shock metabolites” produced by thermotolerant actinomycetes in high-temperature culture. J Antibiot 73, 203–210 (2020). https://doi.org/10.1038/s41429-020-0279-4

Download citation

Search

Quick links