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Subtilase Genes Diversity in the Biogas Digester Microbiota

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Abstract

Biogas digesters contain microbial assemblages that process a mass of extracellular polymeric substances from animal manure and domestic wastewater; however, due to the limitation of available technology in cultivation of majority of the micro-organisms in biogas digesters, the enzymatic potential of these microbial communities remains largely unexplored. In this study, to evaluate subtilase gene diversity in a biogas digester, the partial sequences of the gene were directly amplified from the metagenomic DNA by using consensus-degenerate primers. The desired PCR products were cloned into pGEM-T Easy vector, and thirty positive clones were chose for Polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) analysis, from which thirteen distinguished patterns were obtained and then sequenced. Phylogenetic analysis showed that ten out of the thirteen sequences were related to the subtilase genes in GenBank and were grouped into three families of the subtilases superfamily. The nucleotide sequences analysis through BLAST search revealed that none of the partial genes the authors isolated showed significant similarity against the non-redundant Nucleotide database of NCBI. Meanwhile, the deduced amino acid sequences of ten partial subtilase genes showed moderate identities to the previously identified sequences in GenBank, with a range from 39 to 61%. Collectively, the data indicate that there is a great diversity of subtilase genes in the biogas digester; and may be a rich reservoir for novel subtilase genes.

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Acknowledgments

The authors are very grateful to the reviewers of this article for their careful review, constructive suggestions and language improving. The authors wish to thank Wen-jun Shen for her help in English correcting of the manuscript.

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Authors and Affiliations

  1. Beijing Agro-Biotechnology Research Center, Beijing Academy of Agriculture and Forestry Science, No. 9 Ban Jing Shu Guang Hua Yuan Middle Road, Haidian District, 100097, Beijing, People’s Republic of China

    Xiaojie Cheng, Min Wang, Huawei Liu & Junlian Gao

  2. College of Life Sciences, Capital Normal University, 100048, Beijing, People’s Republic of China

    Xiaojie Cheng & Huawei Liu

  3. Institute of Agricultural Resources and Regional Planning, Chinese Academy of Agricultural Sciences, 100081, Beijing, People’s Republic of China

    Miao Gao & Jianguang Sun

  4. Key Laboratory of Crop Nutrition and Fertilization of Ministry of Agriculture, Chinese Academy of Agricultural Sciences, 100081, Beijing, People’s Republic of China

    Miao Gao & Jianguang Sun

Authors
  1. Xiaojie Cheng
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  2. Miao Gao
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  3. Min Wang
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  4. Huawei Liu
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  5. Jianguang Sun
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  6. Junlian Gao
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Corresponding authors

Correspondence to Jianguang Sun or Junlian Gao.

Additional information

This study was supported by the Key Program of Beijing Municipal Natural Science Foundation (no. 5081001).

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Cheng, X., Gao, M., Wang, M. et al. Subtilase Genes Diversity in the Biogas Digester Microbiota. Curr Microbiol 62, 1542–1547 (2011). https://doi.org/10.1007/s00284-011-9876-6

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  • DOI: https://doi.org/10.1007/s00284-011-9876-6

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