A High-Throughput Screening System Based on Droplet Microfluidics for Glucose Oxidase Gene Libraries.

ABSTRACT: Glucose oxidase (GOx) is an important industrial enzyme that can be optimized for specific applications by mutagenesis and activity-based screening. To increase the efficiency of this approach, we have developed a new ultrahigh-throughput screening platform based on a microfluidic lab-on-chip device that allows the sorting of GOx mutants from a saturation mutagenesis library expressed on the surface of yeast cells. GOx activity was measured by monitoring the fluorescence of water microdroplets dispersed in perfluorinated oil. The signal was generated via a series of coupled enzyme reactions leading to the formation of fluorescein. Using this new method, we were able to enrich the yeast cell population by more than 35-fold for GOx mutants with higher than wild-type activity after two rounds

SUBMITTER: Prodanovic R 

PROVIDER: S-EPMC7287683 | biostudies-literature | 2020 May

REPOSITORIES: biostudies-literature