Caffeine Junkie: an Unprecedented Glutathione S-Transferase-Dependent Oxygenase Required for Caffeine Degradation by Pseudomonas putida CBB5

Supplemental material

Files in this Data Supplement:

• Supplemental file 1 -

  Supplemental methods

  Fig. S1, NdmAB do not form a complex with His₆-NdmCDE

  Fig. S2, oxygen consumption for N₇-demethylation of 7-methylxanthine (A) and Michaelis-Menten plot for kinetic determination for His₆-NdmCDE toward 7-methylxanthine (B)

  Fig. S3, characterization of 7-methyluric acid produced from 7-methylxanthine by single-gene-knockout strains of P. putida CBB5

  Fig. S4, schematic representation of predicted electron flow through the 7-methylxanthine N₇-demethylation components in the caffeine degradation pathway of P. putida CBB5 based on that observed in other RO systems

  Table S1, apparent solubility of NdmCDE expressed in different combinations in E. coli BL21(DE3)

  Table S2, protein accession and GI numbers for NdmCDE gene clusters

  Table S3, PCR primers

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