Journal of Microbiology

Research Support, Non-U.S. Gov't

NOTE] Development of a High-Throughput Screening Method for Recombinant Escherichia coli with Intracellular Dextransucrase Activity: So-Ra Lee , Ah-Rum Yi , Hong-Gyun Lee , Myoung-Uoon Jang , Jung-Mi Park , Nam Soo Han , Tae-Jip Kim; J. Microbiol. 2011;49(2):320-323. Published online May 3, 2011; DOI: https://doi.org/10.1007/s12275-011-1078-1

Abstract: To efficiently engineer intracellular dextransucrase (DSase) expression in Escherichia coli, a high-throughput screening method was developed based on the polymer-forming activity of the enzyme. Recombinant E. coli containing the Leuconostoc citreum DSase (LcDS) gene was grown on Luria-Bertani agar plates, containing 2% sucrose, at 37°C for 8 h. The plates were then evenly overlaid with 0.6% soft agar, containing 1.2 mg/ml D-cycloserine, and incubated at 30°C to allow gradual cell disruption until a dextran polymer grew through the overlaid layer. A significant correlation between dextran size and enzyme activity was established and applied for screening truncated mutants with LcDS activity.

Search