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

作     者：Kenza Samlali Chiara Leal Alves Mara Jezernik Steve C.C.Shih Kenza Samlali;Chiara Leal Alves;Mara Jezernik;Steve C.C.Shih

作者机构：Department of Electrical and Computer EngineeringConcordia UniversityMontréalQCCanada Centre for Applied Synthetic BiologyConcordia UniversityMontréalQCCanada Department of Chemical Engineering and Applied ChemistryUniversity of TorontoTorontoONCanada Department of BiologyConcordia UniversityMontréalQCCanada 

出 版 物：《Microsystems & Nanoengineering》 (微系统与纳米工程（英文）)

年 卷 期：2022年第8卷第6期

页      面：97-112页

核心收录：

学科分类：081203[工学-计算机应用技术] 08[工学] 0835[工学-软件工程] 0812[工学-计算机科学与技术（可授工学、理学学位）] 

基　　金：the Natural Sciences and Engineering Research Council(NSERC) the Fonds de Recherche Nature et technologies(FRQNT) the Canadian Foundation of Innovation(CFI)for funding K.S.thanks the FRQNT B2X&NSERC CREATE SynBioApps program for graduate funding 

主　　题：system fluid culture 

摘      要：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 for4 *** 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.