Discovery of two novel β-glucosidases from an Amazon soil metagenomic library

Authors

  • Jessica C. Bergmann,

    1. Genomic Sciences and Biotechnology Program, Universidade Católica de Brasília, Brasília, Brazil
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  • Ohana Yonara A. Costa,

    1. Genomic Sciences and Biotechnology Program, Universidade Católica de Brasília, Brasília, Brazil
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  • John M. Gladden,

    1. Joint BioEnergy Institute, Emeryville, CA, USA
    2. Biological and Materials Science Center, Sandia National Laboratory, Livermore, CA, USA
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  • Steven Singer,

    1. Joint BioEnergy Institute, Emeryville, CA, USA
    2. Earth Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA, USA
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  • Richard Heins,

    1. Joint BioEnergy Institute, Emeryville, CA, USA
    2. Biological and Materials Science Center, Sandia National Laboratory, Livermore, CA, USA
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  • Patrik D'haeseleer,

    1. Joint BioEnergy Institute, Emeryville, CA, USA
    2. Bioscience and Biotechnology Division, Lawrence Livermore National Laboratory, Livermore, CA, USA
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  • Blake A. Simmons,

    1. Joint BioEnergy Institute, Emeryville, CA, USA
    2. Biological and Materials Science Center, Sandia National Laboratory, Livermore, CA, USA
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  • Betania F. Quirino

    Corresponding author
    1. Genomic Sciences and Biotechnology Program, Universidade Católica de Brasília, Brasília, Brazil
    2. Embrapa-Agroenergy, Brasília, Brazil
    • Correspondence: Betania F. Quirino, Embrapa-Agroenergy, Parque Estação Biológica, s/n; 70.770-901 Brasília, DF, Brazil. Tel.: +55 61 3448 7115; fax: +55 61 3274 3127; e-mails: betania.quirino@embrapa.br; and bfq@uwalumni.com

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Abstract

An Amazon soil microbial community metagenomic fosmid library was functionally screened for β-glucosidase activity. Contig analysis of positive clones revealed the presence of two ORFs encoding novel β-glucosidases, AmBGL17 and AmBGL18, from the GH3 and GH1 families, respectively. Both AmBGL17 and AmBGL18 were functionally identified as β-glucosidases. The enzymatic activity of AmBGL17 was further characterized. AmBGL17 was tested with different substrates and showed highest activity on pNPβG substrate with an optimum temperature of 45 °C and an optimum pH of 6. AmBGL17 showed a Vmax of 116 mM s−1 and Km of 0.30 ± 0.017 mM. This is the first report of β-glucosidases from an Amazon soil microbial community using a metagenomic approach.

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