4.7 Article

A flow cytometer-based whole cell screening toolbox for directed hydrolase evolution through fluorescent hydrogels

Journal

CHEMICAL COMMUNICATIONS
Volume 51, Issue 41, Pages 8679-8682

Publisher

ROYAL SOC CHEMISTRY
DOI: 10.1039/c5cc01791b

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Funding

  1. German Federal Ministry of Education and Research (BMBF) [FKZ: 031A165, FKZ: 031A227F]
  2. Alliance FuPol (Funktionalisierung von Polymeren)

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A high throughput whole cell flow cytometer screening toolbox was developed and validated by identifying improved variants (1.3-7-fold) for three hydrolases (esterase, lipase, cellulase). The screening principle is based on coupled enzymatic reaction using glucose derivatives which yield upon hydrolysis a fluorescent-hydrogel-layer on the surface of E. coli cells.

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