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FIELD Comp.Sciences:Life Science
DATE August 12 (Thu), 2021
TIME 11:00-12:00
PLACE 7323
SPEAKER Kisurb Choe
HOST Hyeon, Changbong
INSTITUTE University of Illinois at Urbana Champaign
TITLE High throughput measurement of fatty acids on microbial colonies by optically guided MALDI-ToF-MS workflow for screening enzyme mutant libraries

Engineering an enzyme often involves creation of many mutants followed by screening to identify a version that meets a goal. The bottleneck for the process is lack of high throughput assay that can directly measure the chemical product of enzyme activity. For example, directed evolution of an enzyme is limited by the number of samples and type of compounds that can be tested by the measurement instrument.

In this talk, I introduce a high throughput mass spectrometry workflow for testing large number of mutants up to 5,000 per day. A custom software, an instrument, and a bioanalytical experiment workflow are developed to create the workflow. The workflow tests microbial colonies grown on Petri dish instead of liquid culture, greatly simplifying and accelerating the chemical screen. Random mutant library of Thioesterase from Umbellularia californica was screened, revealing enzyme variants showing modified substrate specificity towards producing medium chain fatty acids.

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