Adaptive Laboratory Evolution
Adaptive laboratory evolution is a powerful tool to obtain production strains with beneficial mutations. At the Agile BioFoundry, we use three types of adaptive laboratory evolution.
Conventional
Organisms are cultivated in selective conditions, allowing beneficial mutations to arise and become more abundant in the population. Superior performers are isolated and sequenced.
Flow Cytometry
Flow cytometry is a powerful technology to rapidly sort a population based on phenotypes. Flow cytometry and fluorescence-activated cell sorting (FACS) can be applied to sort microbial populations solely based on fluorescence. This can be performed in conjunction with adaptive laboratory evolution or droplet adaptive laboratory evolution to build a diverse population with altered phenotypes.
Droplet Adaptive Laboratory Evolution (dALE)
Droplet adaptive laboratory evolution is a miniaturized approach to adaptive laboratory evolution that can be used to identify performance enhancements that increase production rates.
Droplet adaptive laboratory evolution offers the following advantages compared to traditional adaptive laboratory evolution approaches:
- Cells evolve independently, without negative or positive interactions from other populations.
Miniaturized experiments requires fewer resources - Automation enables Increased experimental efficiency
- Higher genetic diversity and generation numbers achieved in a smaller experimental volume
- A cleaner evolution environment with minimized cross-contamination
- Complete medium refreshment, ensuring optimal growth conditions
- Evolution proceeds from single cells at each dilution step, eliminating unwanted influences from other variants
High-throughput mass-spectrometry-based (HTP-MS) method
We have further optimized high-throughput small-scale evaluation of isolates using a High-Throughput Mass Spectrometry method combined with automation. To date, we have successfully analyzed over 5,000 samples using this system. We have demonstrated that the results obtained from small-scale evaluations—specifically titer, rate, and yield—are reproducible at the bioreactor scale.
USE CASE
Agile BioFoundry researchers used adaptive laboratory evolution to improve the production of muconic acid in Pseudomonas putida KT2440.
Adaptive laboratory evolution was used to identify solutions to overcome a growth defect in a muconate-producing strain of P. putida.
