Science Advances

Supplementary Materials

This PDF file includes:

  • Fig. S1. Scheme for the fluorescent product entrapment strategy and the cell-based FutC and FucD fucosylation assays using FACS.
  • Fig. S2. Analysis of fluorescence retention in various cells.
  • Fig. S3. Optimization of the FACS-based system.
  • Fig. S4. Site-directed mutagenesis and ordered recombination of the mutation site.
  • Fig. S5. Rational selection of candidates for “best hit” from Cα of catalytic key residue and clustering of α helices on substrate-binding sites for CAST and SSM.
  • Fig. S6. LC-MS analyses of the LeX from LacNAc catalyzed by FutA variants.
  • Fig. S7. Steady-state kinetics of wild type, M26, and M32 measured using various substrates.
  • Fig. S8. Structural insight into the improved activity of the best M32 mutant.
  • Table S1. Model screening of FutA(+) cells.
  • Table S2. Specific activities of FutA and selected mutants using LacNAc and lactose as acceptors.
  • Table S3. Activity comparison between the best mutant in the present study and previously reported FutA enzymes.
  • Table S4. Primers used in this study.
  • Table S5. Data collection and refinement statistics.
  • References (44, 45)

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