Discussion

Biosynthesis of high value chemicals by engineered microorganisms is an application of synthetic biology that offers both economic and environmental advantages. This application increases the need for high-throughput screening tools that can facilitate the detection of best performance among a library of designed microbes. For this reason, we are developing a high-throughput, miniaturised Mass Spectrometry tool for profiling synthetic designed libraries in picodroplets (i.e. small aqueous compartments of 300 – 500 pL) at rates of around 100,000 samples/day.

Combining microfluidic-based picodroplet technology for cell encapsulation and sorting, together with Mass Spectroscopy, we aim to rapidly screen, identify and retrieve the best “hits” from synthetic metabolic pathway libraries. To test this new tool, we have designed three libraries for two synthetic metabolic pathways for the amphoteric amino acids: L-2-homoalanaine and p-amino-D-phenylalanine.
Using our screening technology with picodroplet splitting and retrieval will enable recovery of identified ‘hits’ from a replicate library. This InnovateUK-sponsored project aims to recover the high producing hit phenotypes as live bacteria. These hit microbes will then be analysed for their production of the non-natural amino acid and finally the best performing clones will be sequenced and then examined metabolically.

This will enable new scientific understanding, higher throughputs, lower screening costs, shorten design-build-test cycles and thus be of interest to the current MS user base in the synthetic biology and related areas.