Droplet-based microfluidics for high-throughput screening of a metagenomic library for isolation of microbial enzymes

Masahito Hosokawa, Yuri Hoshino, Yohei Nishikawa, Tomotada Hirose, Dong Hyun Yoon, Tetsushi Mori, Tetsushi Sekiguchi, Shuichi Shoji, Haruko Takeyama*


研究成果: Article査読

83 被引用数 (Scopus)


This paper proposes a high-throughput, function-based screening approach of a metagenomic library for isolating novel microbial enzymes by droplet-based microfluidics. We used gel microdroplets (GMDs) dispersed in oil as picoliter-volume reaction vessels for lipolytic enzyme by encapsulating cells in individual GMDs. Using this approach, we monitored the growth of individual cells encapsulated in GMDs and assessed the enzyme reaction activities at the level of an individual GMD. We then applied this method to screen lipolytic enzyme genes from the metagenomic library constructed from soil collected from a quercus serrate forest of Mount Tsukuba, Ibaraki, Japan. In the workflow presented in this study, metagenomic library clones were encapsulated in 100-pL GMDs with a fluorogenic reporter substrate. A total of 67,000 metagenomic library clones can be screened in only 24. h with reduced consumption of reagents (i.e., <10. μL). As a result, we identified a novel lipolytic enzyme, EstT1, belonging to the EstD2 family of esterases and containing a putative signal peptide, which facilitates enzyme export and catalyzation of substrates in the periplasm. Our study demonstrates the potential of microfluidic GMDs as an efficient tool for metagenomic library screening of industrially relevant enzymes with the potential of significantly reducing the cost and time factors involved in successful practical application of microbial enzymes.

ジャーナルBiosensors and Bioelectronics
出版ステータスPublished - 2015 5月 5

ASJC Scopus subject areas

  • バイオテクノロジー
  • 生物理学
  • 生体医工学
  • 電気化学


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