Simple discovery of bacterial biocatalysts from environmental samples through functional metaproteomics.
Microbiome
; 5(1): 28, 2017 03 03.
Article
em En
| MEDLINE
| ID: mdl-28253936
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.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Proteômica
/
Escherichia coli
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Metabolismo dos Lipídeos
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Recuperação e Remediação Ambiental
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Metagenômica
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Lipase
Idioma:
En
Revista:
Microbiome
Ano de publicação:
2017
Tipo de documento:
Article
País de afiliação:
Alemanha