Skip to main content
SpringerLink
Log in

An integrated high throughput strategy to screen mutants of Paenibacillus polymyxa with high polymyxin E-productivity

  • Original Paper
  • Published:
World Journal of Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

An integrated high-throughput screening (HTS) strategy was developed to screen large numbers of polymyxin E-producing mutants of Paenibacillus polymyxa. Various types of mutants were transferred onto the surfaces of solidified agar in 96-well microtiter plates, and then inoculated to 96-deep-well microtiter plates for micro-cultivation. The culture conditions were optimized for the production of polymyxin E. The supernatants from the micro-culture plates were transferred to 96-well bioassay microtiter plates containing Escherichia coli JM109 for high-throughput bioassay. By using this high-throughput screening (HTS) procedure, one best producer P. polymyxa PE 5.808 was identified from a large NTG mutated library with about 5,000 isolates. The volumetric productivity of polymyxin E of P. polymyxa PE 5.808 was 1,200 μg/ml in shake flasks, about 140% improvement compared with that of the wild type strain.

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

  • Bragulat MR, Aberca ML, Cabanes FJ (2001) An easy screening method for fungi-producing ochratoxin A in pure culture. Int J Food Microbiol 71:139–144

    Article  CAS  Google Scholar 

  • Casey JT, O’Celirigh C, Walsh PK, O’Shea DG (2004) Development of a robust microtiter plate-based assay method for assessment of bioactivity. J Microbiol Methods 58:327–334

    Article  CAS  Google Scholar 

  • Devienne KF, Raddi MSG (2002) Screening for antimicrobial activity of natural products using a microplate photometer. Braz J Microbiol 33:166–168

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Duetz WA, Witholt B (2001) Effectiveness of orbital shaking for the aeration of suspended bacterial cultures in square-deepwell microtiter plates. Biochem Eng J 7:113–115

    Article  CAS  Google Scholar 

  • John GT, Klimant I, Wittmann C (2003) Integrated optical sensing of dissolved oxygen in microtiter plates: a novel tool for microbial cultivation. Biotechnol Bioeng 81:829–836

    Article  CAS  Google Scholar 

  • Johnsen AR, Bendixen K, Karlson U (2002) Detection of microbial growth on polycyclic aromatic hydrocarbons in microtiter plates by using the respiration indicator WST-1. Appl Environ Microbiol 68(6):2683–2689

    Article  CAS  Google Scholar 

  • Kline T, Holub D, Therrien J, Leung T, Ryckman D (2001) Synthesis and characterization of the colistin peptide polymyxin E1 and related antimicrobial peptides. J Peptide Res 57:175–187

    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 carassa bioassay. J Microbiol Methods 40:99–104

    Article  CAS  Google Scholar 

  • Renard L, Gicquel M, Laurentie M, Sanders P (1996) Bactericidal effect of polymyxin E on Escherichia coli. model and simulation or the pharmacokinetic-pharmacodynamic relation for prediction of efficacy in veterinary antibiotic therapy. Vet Res 27(1):23–32

    CAS  Google Scholar 

  • Roveta S, Schito AM, Marchese A, Schito GC (2005) Microbiological rationale for the utilization of prulifloxacin, a new fluoroquinolone, in the eradication of serious infections caused by Pseudomonas aeruginosa. Int J Antimicrob Agents 26:366–372

    Google Scholar 

  • Smedsgaard J (1997) Micro-scale extraction procedure for standardized screening of fungal metabolite production in cultures. J Chromatogr A 760:264–270

    Article  CAS  Google Scholar 

  • Stepanovic S, Vukovic D, Dakic I, Savic B, Svabic-Vlahovic M (2000) A modified microtiter-plate test quantification of Syaphylococcal biolfilm formation. J Microbiol Methods 40:175–179

    Article  CAS  Google Scholar 

  • Xu Z-n, Shen W-h, Chen X-y, Lin J-p, Cen P-l (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

    Google Scholar 

Download references

Acknowledgement

This work was financially supported by the Ministry of Science and Technology of China (National Basic Research Program of China, 2007CB707805; National “863” Research Program of China, 2006AA02Z239).

Author information

Authors and Affiliations

  1. School of Bioscience and Bioengineering, Southern China University of Technology, Guangzhou, 510640, China

    Jufang Wang & Xiaoning Wang

  2. Institute of Bioengineering, Department of Chemical and Biochemical Engineering, Zhejiang University, Hangzhou, 310027, P.R. China

    Yinying Tao, Zhinan Xu, Rong Xu & Peilin Cen

Authors
  1. Jufang Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yinying Tao
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Zhinan Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Rong Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Peilin Cen
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Xiaoning Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Zhinan Xu.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wang, J., Tao, Y., Xu, Z. et al. An integrated high throughput strategy to screen mutants of Paenibacillus polymyxa with high polymyxin E-productivity. World J Microbiol Biotechnol 24, 1885–1891 (2008). https://doi.org/10.1007/s11274-008-9686-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11274-008-9686-6

Keywords

Search

Navigation