Highlights
• With single-cell Raman spectroscopy, characteristic peaks correlated with diacylglycerol acyltransferase activity can be identified
• Rapid screening of functional enzyme libraries can be accomplished on FlowRACS®, High-throughput Raman-activated Flow Cytometer
• With FlowRACS®, 3 previously reported strong genes and 2 novel weak genes were obtained in a 10-minute run,compared to months of work using conventional approaches
Abstract
Diacylglycerol acyltransferase (DGAT) is a key enzyme catalyzing the final step of lipid synthesis in human, animals, and plants, and is the only rate-limiting enzyme. Using FlowRACS®, label-free,high-throughput, high-accuracy, non-destructive sorting of DGAT mutant libraries can be achieved at single-cell resolution. In this experiment, from a library of candidate DGAT genes derived from Micrococcus, three previously reported strong genes and two never-reported weak genes were obtained by FlowRACS® in a10-minute run, whereas the screening and characterization of these three strong functional genes with traditional methods would have taken several months. In conclusion, FlowRACS® provides a powerful tool for mining highly efficient enzymes.
