Scheduled maintenance This site will be offline from 13:30 to 15:00 CEST on Monday, 21 October 2024 in order to be moved to a new infrastructure. We apologise for the inconvenience. Please check the Slack channel #dc-system-status for updates. The development team can be contacted at datacentre@scilifelab.se if you have any question.

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

Sjostrom SL, Bai Y, Huang M, Liu Z, Nielsen J, Joensson HN, Andersson Svahn H

Lab Chip 14 (4) 806-813 [2014-02-21; online 2013-12-25]

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 10(5) 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.

Affiliated researcher

PubMed 24366236

DOI 10.1039/c3lc51202a

Crossref 10.1039/c3lc51202a


Publications 9.5.0