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Response surface optimization of a method for extracting extracellular polymeric substances (EPS) from subaerial biofilms on rocky substrata

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Abstract

The aim of the present study was to optimize a protocol for extracting extracellular polymeric substances (EPS) from biofilms on rocky substrata, as the EPS matrix is considered key to understanding the biofilm mode of life. For this purpose, we tested the extraction efficacy of NaOH and H2SO4 at different concentrations, temperatures and times for obtaining EPS from multi-species subaerial biofilms grown on granite blocks under laboratory conditions. Two experimental designs (Box-Behnken design and full factorial design) were used in testing each extractant. The extraction efficiency was determined by analysing the carbohydrate, protein and DNA contents of the extracts obtained. H2SO4 proved unsuitable as an extractant as it caused excessive cell lysis. However, response surface optimization of NaOH-mediated extraction enabled cell lysis to be minimized. Confirmation experiments were performed under the optimal conditions established and a protocol for extracting EPS is proposed, yielding the first quantitative data on EPS extracted from subaerial biofilms developed on rocky substrata.

Development of a method for extracting EPS from subaerial biofilms on rocky substrata

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Acknowledgments

The first author was financially supported by “Programa de axudas predoutorais financiadas polos grupos de investigación da Universidade de Santiago de Compostela no marco do Estatuto do Persoal de Investigación en Formación” through PhD grant 2011/AX461.

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Authors and Affiliations

  1. Departamento de Edafoloxía e Química Agrícola, Facultade de Farmacia, Universidade de Santiago de Compostela, Campus Vida, 15782, Santiago de Compostela, Spain

    Daniel Vázquez-Nion, María Echeverri, Benita Silva & Beatriz Prieto

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  1. Daniel Vázquez-Nion
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  2. María Echeverri
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  3. Benita Silva
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  4. Beatriz Prieto
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Correspondence to Daniel Vázquez-Nion.

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Daniel Vázquez-Nion declares that he has no conflict of interest. María Echeverri declares that she has no conflict of interest. Benita Silva declares that she has no conflict of interest. Beatriz Prieto declares that she has no conflict of interest. This article does not contain any studies with human participants or animals performed by any of the authors. All authors have seen and agree with the contents of the manuscript.

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Vázquez-Nion, D., Echeverri, M., Silva, B. et al. Response surface optimization of a method for extracting extracellular polymeric substances (EPS) from subaerial biofilms on rocky substrata. Anal Bioanal Chem 408, 6369–6379 (2016). https://doi.org/10.1007/s00216-016-9752-0

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