Jump to main content
Jump to site search

Issue 4, 2014
Previous Article Next Article

High-throughput screening for industrial enzyme production hosts by droplet microfluidics

Author affiliations

Abstract

A high-throughput method for single cell screening by microfluidic droplet sorting is applied to a whole-genome mutated yeast cell library yielding improved production hosts of secreted industrial enzymes. The sorting method is validated by enriching a yeast strain 14 times based on its α-amylase production, close to the theoretical maximum enrichment. Furthermore, a 105 member yeast cell library is screened yielding a clone with a more than 2-fold increase in α-amylase production. The increase in enzyme production results from an improvement of the cellular functions of the production host in contrast to previous droplet-based directed evolution that has focused on improving enzyme protein structure. In the workflow presented, enzyme producing single cells are encapsulated in 20 pL droplets with a fluorogenic reporter substrate. The coupling of a desired phenotype (secreted enzyme concentration) with the genotype (contained in the cell) inside a droplet enables selection of single cells with improved enzyme production capacity by droplet sorting. The platform has a throughput over 300 times higher than that of the current industry standard, an automated microtiter plate screening system. At the same time, reagent consumption for a screening experiment is decreased a million fold, greatly reducing the costs of evolutionary engineering of production strains.

Graphical abstract: High-throughput screening for industrial enzyme production hosts by droplet microfluidics

Back to tab navigation

Supplementary files

Publication details

The article was received on 23 Oct 2013, accepted on 28 Nov 2013 and first published on 28 Nov 2013


Article type: Paper
DOI: 10.1039/C3LC51202A
Lab Chip, 2014,14, 806-813

  •   Request permissions

    High-throughput screening for industrial enzyme production hosts by droplet microfluidics

    S. L. Sjostrom, Y. Bai, M. Huang, Z. Liu, J. Nielsen, H. N. Joensson and H. Andersson Svahn, Lab Chip, 2014, 14, 806
    DOI: 10.1039/C3LC51202A

Search articles by author

Spotlight

Advertisements