The E. coli FutA(+) cells expressing FuctA were mixed with FutA(−) cells harboring empty plasmid pUC18 at ratios of 1:10, 1:100, 1:1000, and 1:10,000. The cell mixtures were reacted with two fluorogenic substrates at 37°C for 30 min, and the excess acceptor sugars were removed by washing the cells with LB media and PBS (pH 7.4). Next, the samples were sorted by flow cytometry, and single E. coli cells with the top 0.5% of blue fluorescence intensity were collected into 1.5-ml tubes containing 0.5 ml of super optimal broth with catabolite repression (SOC) medium. After growing the collected cells on agar plates, negative and positive colonies were identified by individual bacterial colony PCR with the universal primers M13F(−47)/M13R(−48). The enrichment factors were calculated on the basis of the positive ratio values from before and after sorting.

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