Title |
Simple discovery of bacterial biocatalysts from environmental samples through functional metaproteomics
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Published in |
Microbiome, March 2017
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DOI | 10.1186/s40168-017-0247-9 |
Pubmed ID | |
Authors |
Premankur Sukul, Sina Schäkermann, Julia E. Bandow, Anna Kusnezowa, Minou Nowrousian, Lars I. Leichert |
Abstract |
Bacterial biocatalysts play a key role in our transition to a bio-based, post-petroleum economy. However, the discovery of new biocatalysts is currently limited by our ability to analyze genomic information and our capacity of functionally screening for desired activities. Here, we present a simple workflow that combines functional metaproteomics and metagenomics, which facilitates the unmediated and direct discovery of biocatalysts in environmental samples. To identify the entirety of lipolytic biocatalysts in a soil sample contaminated with used cooking oil, we detected all proteins active against a fluorogenic substrate in sample's metaproteome using a 2D-gel zymogram. Enzymes' primary structures were then deduced by tryptic in-gel digest and mass spectrometry of the active protein spots, searching against a metagenome database created from the same contaminated soil sample. We then expressed one of the novel biocatalysts heterologously in Escherichia coli and obtained proof of lipolytic activity. |
X Demographics
Geographical breakdown
Country | Count | As % |
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Canada | 1 | 17% |
Unknown | 5 | 83% |
Demographic breakdown
Type | Count | As % |
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Members of the public | 3 | 50% |
Scientists | 2 | 33% |
Practitioners (doctors, other healthcare professionals) | 1 | 17% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
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Brazil | 2 | 3% |
Japan | 1 | 1% |
Unknown | 65 | 96% |
Demographic breakdown
Readers by professional status | Count | As % |
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Student > Ph. D. Student | 18 | 26% |
Researcher | 12 | 18% |
Student > Bachelor | 9 | 13% |
Student > Master | 6 | 9% |
Student > Doctoral Student | 3 | 4% |
Other | 12 | 18% |
Unknown | 8 | 12% |
Readers by discipline | Count | As % |
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Agricultural and Biological Sciences | 21 | 31% |
Biochemistry, Genetics and Molecular Biology | 17 | 25% |
Environmental Science | 5 | 7% |
Engineering | 3 | 4% |
Chemistry | 3 | 4% |
Other | 7 | 10% |
Unknown | 12 | 18% |