Droplet-based microfluidic platform for heterogeneous enzymatic assays. 2013

Chieh Chang, and Jess Sustarich, and Rajiv Bharadwaj, and Aarthi Chandrasekaran, and Paul D Adams, and Anup K Singh
Technology Division, Joint BioEnergy Institute, Emeryville, California 94608, United States.

Heterogeneous enzymatic reactions are used in many industrial processes including pulp and paper, food, and biofuel production. Industrially-relevant optimization of the enzymes used in these processes requires assaying them with insoluble substrates. However, platforms for high throughput heterogeneous assays do not exist thereby severely increasing the cost and time of enzyme optimization, or leading to the use of assays with soluble substrates for convenient, but non-ideal, optimization. We present an innovative approach to perform heterogeneous reactions in a high throughput fashion using droplet microfluidics. Droplets provide a facile platform for heterogeneous reactions as internal recirculation allows rapid mixing of insoluble substrates with soluble enzymes. Moreover, it is easy to generate hundreds or thousands of picoliter droplets in a small footprint chip allowing many parallel reactions. We validate our approach by screening combinations of cellulases with real-world insoluble substrates, and demonstrate that the chip-based screening is in excellent agreement with the conventional screening methods, while offering advantages of throughput, speed and lower reagent consumption. We believe that our approach, while demonstrated for a biofuel application, provides a generic platform for high throughput monitoring of heterogeneous reactions.

UI MeSH Term Description Entries
D002480 Cellulase An endocellulase with specificity for the hydrolysis of 1,4-beta-glucosidic linkages in CELLULOSE, lichenin, and cereal beta-glucans. Endo-1,4-beta-Glucanase,Cellulysin,Endoglucanase,Endoglucanase A,Endoglucanase C,Endoglucanase E,Endoglucanase IV,Endoglucanase Y,beta-1,4-Glucan-4-Glucanohydrolase,Endo 1,4 beta Glucanase,beta 1,4 Glucan 4 Glucanohydrolase
D001617 beta-Glucosidase An exocellulase with specificity for a variety of beta-D-glycoside substrates. It catalyzes the hydrolysis of terminal non-reducing residues in beta-D-glucosides with release of GLUCOSE. Cellobiases,Amygdalase,Cellobiase,Emulsion beta-D-Glucosidase,Gentiobiase,Emulsion beta D Glucosidase,beta Glucosidase,beta-D-Glucosidase, Emulsion
D043366 Cellulose 1,4-beta-Cellobiosidase An exocellulase with specificity for the hydrolysis of 1,4-beta-D-glucosidic linkages in CELLULOSE and cellotetraose. It catalyzes the hydrolysis of terminal non-reducing ends of beta-D-glucosides with release of CELLOBIOSE. Exo-Cellobiohydrolase,1,4-beta-D-Glucan Cellobiohydrolase,Beta-Glucancellobiohydrolase,CEL1 Protein,CbhI Protein,Cellobiohydrolase,Cellobiohydrolase A,Cellobiohydrolase I,Cellobiohydrolase II,Exoglucanase II,1,4 beta D Glucan Cellobiohydrolase,1,4-beta-Cellobiosidase, Cellulose,Beta Glucancellobiohydrolase,Cellobiohydrolase, 1,4-beta-D-Glucan,Cellulose 1,4 beta Cellobiosidase,Exo Cellobiohydrolase
D046210 Microfluidic Analytical Techniques Methods utilizing the principles of MICROFLUIDICS for sample handling, reagent mixing, and separation and detection of specific components in fluids. Microfluidic Analysis,Analyses, Microfluidic,Analysis, Microfluidic,Analytical Technique, Microfluidic,Analytical Techniques, Microfluidic,Microfluidic Analyses,Microfluidic Analytical Technique,Technique, Microfluidic Analytical,Techniques, Microfluidic Analytical

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