Simple discovery of bacterial biocatalysts from environmental samples through functional metaproteomics
- 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 \(\textit {Escherichia coli}\) and obtained proof of lipolytic activity.
| Author: | Premankur SukulGND, Sina SchäkermannORCiDGND, Julia BandowORCiDGND, Anna KusnezowaGND, Minou NowrousianORCiDGND, Lars I. LeichertORCiDGND |
|---|---|
| URN: | urn:nbn:de:hbz:294-58800 |
| DOI: | https://doi.org/10.1186/s40168-017-0247-9 |
| Parent Title (English): | Microbiome |
| Document Type: | Article |
| Language: | English |
| Date of Publication (online): | 2018/07/10 |
| Date of first Publication: | 2017/03/03 |
| Publishing Institution: | Ruhr-Universität Bochum, Universitätsbibliothek |
| Tag: | Open Access Fonds Biocatalyst; Lipase; Metagenomics; Metaproteomics; Zymogram |
| Volume: | 5 |
| Issue: | 1 |
| First Page: | 28 |
| Note: | Article Processing Charge funded by the Deutsche Forschungsgemeinschaft (DFG) and the Open Access Publication Fund of Ruhr-Universität Bochum. |
| Institutes/Facilities: | Institut für Biochemie und Pathobiochemie, Abteilung Biochemie der Mikroorganismen |
| Dewey Decimal Classification: | Naturwissenschaften und Mathematik / Biowissenschaften, Biologie, Biochemie |
| open_access (DINI-Set): | open_access |
| faculties: | Medizinische Fakultät |
| Licence (English): |  Creative Commons - CC BY 4.0 - Attribution 4.0 International |
