Two promising alkaline β-glucosidases isolated by functional metagenomics from agricultural soil, including one showing high tolerance towards harsh detergents, oxidants and glucose

  • Sophie Biver
  • Aurore Stroobants
  • Daniel Portetelle
  • Micheline Vandenbol
Biocatalysis

Abstract

New β-glucosidase activities were identified by screening metagenomic libraries constructed with DNA isolated from the topsoil of a winter wheat field. Two of the corresponding proteins, displaying an unusual preference for alkaline conditions, were selected for purification by Ni-NTA chromatography. AS-Esc6, a 762-amino-acid enzyme belonging to glycoside hydrolase family 3, proved to be a mesophilic aryl-β-glucosidase with maximal activity around pH 8 and 40 °C. A similar pH optimum was found for AS-Esc10, a 475-amino-acid GH1-family enzyme, but this enzyme remained significantly active across a wider pH range and was also markedly more stable than AS-Esc6 at pH greater than 10. AS-Esc10 was found to degrade cellobiose and diverse aryl glycosides, with an optimal temperature of 60 °C and good stability up to 50 °C. Unlike AS-Esc6, which showed a classically low inhibitory constant for glucose (14 mM), AS-Esc10 showed enhanced activity in the presence of molar concentrations of glucose. AS-Esc10 was highly tolerant to hydrogen peroxide and also to sodium dodecyl sulfate, this being indicative of kinetic stability. This unique combination of properties makes AS-Esc10 a particularly promising candidate whose potential in biotechnological applications is worth exploring further.

Keywords

Alkaline enzyme β-Glucosidase Detergent Functional metagenomics Glucose tolerance 

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Copyright information

© Society for Industrial Microbiology and Biotechnology 2014

Authors and Affiliations

  • Sophie Biver
    • 1
  • Aurore Stroobants
    • 1
  • Daniel Portetelle
    • 1
  • Micheline Vandenbol
    • 1
  1. 1.Microbiology and Genomics Unit, Gembloux Agro-Bio TechUniversity of LiègeGemblouxBelgium

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