High-Throughput Screening System and Its Biotechnological Applications

Novel enzyme and/or metabolic pathway screening techniques from various environmental resources and its biotechnological applications have widely expanded in the past few years. However, it is difficult to screen new enzymes due to limited assay protocol, expensive substrate, library, low sensitivity, low activity of target enzymes in host strain, and time-consuming and laborious process. Therefore it was difficult for mass screening of target enzymes from various resources. We had been developed the HTS system for enzyme screening from various environmental resources including metagenome. This HTS system composed with Robotic Integration System, JANUS Liquid Handling System, VICTOR3 Multilabel Microplate Reader, K3 Colony Picking System, Automated Incubator, Carousel, Lidding/Delidding System, Bar Code Reader, FACSAria High-speed Flow Cytometer. Finally optimized screening methods for high throughput system led to over 10⁵ clone active verification per day were available.

In this study, we developed mass screening methods for various enzymes from metagenomic libraries using HTS system based on robot. We could new enzymes such as cellobiohydrolases, glycosyltransferases, BVMO, cold-adapted esterase and deoxyribose 5-phosphate aldolase(DERA)  based on fluorescence intensity. These new strategies combined with HTS system could screen various new enzyems/metabolic pathways more fast, sensitive and easy than previously reported screening methods. This approach would be applied for other useful enzyme and metabolic pathway screening from metagenomic resources.