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Droplet digital microfluidic system for screening filamentous fungi based on enzymatic activity

Microsystems & Nanoengineering 2022年第6期8卷 97-112页

作者： kenza samlali Chiara Leal Alves Mara Jezernik Steve C.C.Shih Department of Electrical and Computer Engineering Concordia UniversityMontréalQCCanada Centre for Applied Synthetic Biology Concordia UniversityMontréalQCCanada Department of Chemical Engineering and Applied Chemistry University of TorontoTorontoONCanada Department of Biology Concordia UniversityMontréalQCCanada

Fungal cell-wall-degrading enzymes have great utility in the agricultural and food *** cell-wall-degrading enzymes are known to have functions that can help defend against pathogenic *** existing methods used to discover these enzymes are not well adapted to fungi culture and morphology,which prevents the proper evaluation of these *** report the first droplet-based microfluidic method capable of long-term incubation and low-voltage conditions to sort filamentous fungi inside nanoliter-sized *** new method was characterized and validated in solid-phase media based on colloidal chitin such that the incubation of single spores in droplets was possible over multiple days(2–4 days)and could be sorted without droplet *** long-term culture,we examined the activity of cell-wall-degrading enzymes produced by fungi during solid-state droplet fermentation using three highly sensitive fluorescein-based *** also used the low-voltage droplet sorter to select clones with highly active cell-wall-degrading enzymes,such as chitinases,β-glucanases,and β-Nacetylgalactosaminidases,from a filamentous fungi droplet library that had been incubated for>4 *** new system is portable,affordable for any laboratory,and user-friendly compared to classical droplet-based microfluidic *** propose that this system will be useful for the growing number of scientists interested in fungal microbiology who are seeking high-throughput methods to incubate and sort a large library of fungal cells.

关键词： system fluid culture

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