2016年2月
Culture-independent method for identification of microbial enzyme-encoding genes by activity-based single-cell sequencing using a water-in-oil microdroplet platform
Scientific Report
- 巻
- 6
- 号
- 開始ページ
- 22259
- 終了ページ
- 記述言語
- 英語
- 掲載種別
- 研究論文(学術雑誌)
- DOI
- 10.1038/srep22259
- 出版者・発行元
- NATURE PUBLISHING GROUP
Environmental microbes are a great source of industrially valuable enzymes with potent and unique catalytic activities. Unfortunately, the majority of microbes remain unculturable and thus are not accessible by culture-based methods. Recently, culture-independent metagenomic approaches have been successfully applied, opening access to untapped genetic resources. Here we present a methodological approach for the identification of genes that encode metabolically active enzymes in environmental microbes in a culture-independent manner. Our method is based on activity-based single-cell sequencing, which focuses on microbial cells showing specific enzymatic activities. First, at the single-cell level, environmental microbes were encapsulated in water-in-oil microdroplets with a fluorogenic substrate for the target enzyme to screen for microdroplets that contain microbially active cells. Second, the microbial cells were recovered and subjected to whole genome amplification. Finally, the amplified genomes were sequenced to identify the genes encoding target enzymes. Employing this method, we successfully identified 14 novel beta-glucosidase genes from uncultured bacterial cells in marine samples. Our method contributes to the screening and identification of genes encoding industrially valuable enzymes.
- リンク情報
-
- DOI
- https://doi.org/10.1038/srep22259
- PubMed
- https://www.ncbi.nlm.nih.gov/pubmed/26915788
- Web of Science
- https://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=JSTA_CEL&SrcApp=J_Gate_JST&DestLinkType=FullRecord&KeyUT=WOS:000370873000001&DestApp=WOS_CPL
- 共同研究・競争的資金等の研究課題
- 液滴を利用した微生物培養を介さない酵素遺伝子取得法の確立とその実践
- Scopus
- https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=84959387605&origin=inward 本文へのリンクあり
- Scopus Citedby
- https://www.scopus.com/inward/citedby.uri?partnerID=HzOxMe3b&scp=84959387605&origin=inward
- ID情報
-
- DOI : 10.1038/srep22259
- ISSN : 2045-2322
- eISSN : 2045-2322
- PubMed ID : 26915788
- SCOPUS ID : 84959387605
- Web of Science ID : WOS:000370873000001