Skip to main content

Advertisement

Log in

Identification of avermectin-high-producing strains by high-throughput screening methods

  • Methods
  • Published:
Applied Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

Avermectins produced by Streptomyces avermitilis are potent against a broad spectrum of nematode and arthropod parasites with low-level side effects on the host organisms. This study was designed to investigate a high-throughput screening strategy for the efficient identification of avermectin high-yield strains. The production protocol was miniaturized in 96 deep-well microplates. UV absorbance at 245 nm was used to monitor avermectin production. A good correlation between fermentation results in both 96 deep-well microplates and conventional Erlenmeyer flasks was observed. With this protocol, the production of avermectins was determined in less than 10 min for a full plate without compromising accuracy. The high-yield strain selected through this protocol was also tested in 360 m3 batch fermentation with 1.6-fold improved outcome. Thus, the development of this protocol is expected to accelerate the selection of superior avermectin-producing strains.

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

Access this article

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

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

Instant access to the full article PDF.

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

Similar content being viewed by others

Explore related subjects

Discover the latest articles and news from researchers in related subjects, suggested using machine learning.

References

  • Askal HF, Refaat IH, Darwish IA, Marzouq MA (2008) A selective spectrophotometric method for determination of rosoxacin antibiotic using sodium nitroprusside as a chromogenic reagent. Spectrochim Acta A Mol Biomol Spectrosc 69:1287–1291

    Article  Google Scholar 

  • Buchs J (2001) Introduction to advantages and problems of shaken cultures. Biochem Eng J 7:91–98

    Article  CAS  Google Scholar 

  • Burg RW, Miller BM, Baker EE, Birnbaum J, Currie SA, Hartman R, Kong YL, Monaghan RL, Olson G, Putter I, Tunac JB, Wallick H, Stapley EO, Oiwa R, Omura S (1979) Avermectins, new family of potent anthelmintic agents: producing organism and fermentation. Antimicrob Agents Chemother 15:361–367

    CAS  Google Scholar 

  • Curdova E, Jechova V, Zima J, Vanek Z (1989) The effect of inorganic phosphate on the production of avermectin in Streptomyces avermitilis. J Basic Microbiol 29:341–346

    Article  CAS  Google Scholar 

  • del Cardayré SB (2005) Developments in strain improvement technology: evolutionary engineering of industrial microorganisms through gene, pathway, and genome shuffling. In: Zhang L, Demain AL (eds) Natural products: drug discovery and therapeutics medicines. Humana, Totowa, NJ, pp 33–56

    Google Scholar 

  • Dobson LF, O'Cleirigh CC, O'Shea DG (2008) The influence of morphology on geldanamycin production in submerged fermentations of Streptomyces hygroscopicus var. geldanus. Appl Microbiol Biotechnol 79:859–866

    Article  CAS  Google Scholar 

  • Du CY, Zhang YP, Li Y, Cao Z (2007) Novel redox potential-based screening strategy for rapid isolation of Klebsiella pneumoniae mutants with enhanced 1,3-propanediol-producing capability. Appl Environ Microbiol 73:4515–4521

    Article  CAS  Google Scholar 

  • du Toit EA, Rautenbach M (2000) A sensitive standardised micro-gel well diffusion assay for the determination of antimicrobial activity. J Microbiol Methods 42:159–165

    Article  Google Scholar 

  • Duetz WA, Ruedi L, Hermann R, O'Connor K, Buchs J, Witholt B (2000) Methods for intense aeration, growth, storage, and replication of bacterial strains in microtiter plates. Appl Environ Microbiol 66:2641–2646

    Article  CAS  Google Scholar 

  • Gao H, Liu M, Liu JT, Dai HQ, Zhou XL, Liu XY, Zhuo Y, Zhang WQ, Zhang L (2009) Medium optimization for the production of avermectin B1a by Streptomyces avermitilis 14–12A using response surface methodology. Bioresour Technol 100:4012–4016

    Article  CAS  Google Scholar 

  • Hermann R, Lehmann M, Buchs J (2003) Characterization of gas–liquid mass transfer phenomena in microtiter plates. Biotechnol Bioeng 81:178–186

    Article  CAS  Google Scholar 

  • Hwang YS, Kim ES, Biro S, Choi CY (2003) Cloning and analysis of a DNA fragment stimulating avermectin production in various Streptomyces avermitilis strains. Appl Environ Microbiol 69:1263–1269

    Article  CAS  Google Scholar 

  • Ikeda H, Omura S (1995) Control of avermectin biosynthesis in Streptomyces avermitilis for the selective production of a useful component. J Antibiot (Tokyo) 48:549–562

    CAS  Google Scholar 

  • Ikeda H, Omura S (1997) Avermectin biosynthesis. Chem Rev 97:2591–2610

    Article  CAS  Google Scholar 

  • Isett K, George H, Herber W, Amanullah A (2007) Twenty-four-well plate miniature bioreactor high-throughput system: assessment for microbial cultivations. Biotechnol Bioeng 98:1017–1028

    Article  CAS  Google Scholar 

  • Kumar MS, Kumar PM, Sarnaik HM, Sadhukhan AK (2000) A rapid technique for screening of lovastatin-producing strains of Aspergillus terreus by agar plug and Neurospora crassa bioassay. J Microbiol Methods 40:99–104

    Article  CAS  Google Scholar 

  • Lee J, Hwang Y, Kim S, Kim E, Choi C (2000) Effect of a global regulatory gene, afsR2, from Streptomyces lividans on avermectin production in Streptomyces avermitilis. J Biosci Bioeng 89:606–608

    Article  CAS  Google Scholar 

  • Sams R (1993) Chemical assay of avermectins by high performance liquid chromatography with fluorescence detection. Vet Parasitol 48:59–66

    Article  CAS  Google Scholar 

  • Tyo KE, Zhou H, Stephanopoulos GN (2006) High-throughput screen for poly-3-hydroxybutyrate in Escherichia coli and Synechocystis sp. strain PCC6803. Appl Environ Microbiol 72:3412–3417

    Article  CAS  Google Scholar 

  • Xu ZN, Shen WH, Chen XY, Lin JP, Cen PL (2005) A high-throughput method for screening of rapamycin-producing strains of Streptomyces hygroscopicus by cultivation in 96-well microtiter plates. Biotechnol Lett 27:1135–1140

    Article  CAS  Google Scholar 

  • Yu H, Tyo K, Alper H, Klein-Marcuschamer D, Stephanopoulos G (2008) A high-throughput screen for hyaluronic acid accumulation in recombinant Escherichia coli transformed by libraries of engineered sigma factors. Biotechnol Bioeng 101:788–796

    Article  CAS  Google Scholar 

  • Zhang L (2005a) Integrated approaches for discovering novel drugs from microbial natural products. In: Zhang L, Demain AL (eds) Natural products: drug discovery and therapeutics medicines. Humana, New Jersey, pp 33–56

    Google Scholar 

  • Zhang L (2005b) Screening for synergistic compounds. PCT Patent WO 2005/051303

  • Zhang L, An R, Wang J, Sun N, Zhang S, Hu J, Kuai J (2005) Exploring novel bioactive compounds from marine microbes. Curr Opin Microbiol 11:655–662

    Google Scholar 

  • Zhang L, Yan K, Zhang Y, Bian J, Zheng C, Sun H, Chen Z, Sun N, Song Y, Zhuo Y, You J et al (2007) High-throughput synergy screening identifies microbial metabolites as combination agents for the treatment of fungal infections. PNAS 104:4606–4611

    Article  CAS  Google Scholar 

  • Zimmermann HF, John GT, Trauthwein H, Dingerdissen U, Huthmacher K (2003) Rapid evaluation of oxygen and water permeation through microplate sealing tapes. Biotechnol Prog 19:1061–1063

    Article  CAS  Google Scholar 

Download references

Acknowledgments

We thank Arnold Demain and Elizabeth Ashforth for their critical reading of the manuscript and helpful discussions. This work was supported in part by grants from National Natural Science Foundation of China (No 30700015), National 863 project (2006AA09Z402 and 2007AA09Z443), and Chinese Academy of Sciences Innovation Projects O62A131BB4, National Key Technology R&D Program 2007BAI26B02, the National Science & Technology Pillar Program (No. 200703295000-02), Important National Science & Technology Specific Projects (No. 2008ZX09401-05), Science and Technology Planning Project of Guangdong Province, China (No. 2006A50103001). L.Z. was an awardee for Hundred Talents Program.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Lixin Zhang.

Additional information

Hong Gao, Mei Liu, and Xianlong Zhou contributed equally to this work.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Gao, H., Liu, M., Zhou, X. et al. Identification of avermectin-high-producing strains by high-throughput screening methods. Appl Microbiol Biotechnol 85, 1219–1225 (2010). https://doi.org/10.1007/s00253-009-2345-5

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00253-009-2345-5

Keywords