FIG. 2.
Deletion analysis of the 5.2-kb BamHI-EcoRI fragment carrying ferBA (A) and the restriction map of the 7.0-kb EcoRI fragment carrying ferB2 and ligW (B). The ferulic acid transformation activity and the ability to generate vanillin from ferulic acid of E. coli JM109 harboring each plasmid are presented on the right-hand side of panel A. Small arrows indicate the direction of transcription from the lac promoter. Triangles indicate the positions of the Kmr gene insertion of the ferA mutant (FAK), ferB mutant (FBK), and ferB2 mutant (FB2K). Double-headed arrows indicate the probes used for Southern hybridization analysis of the fer gene mutants. The following genes (and their products) were used: ferA, feruloyl-CoA synthetase; ferB and ferB2, feruloyl-CoA hydratase/lyase; ligW, 5-carboxyvanillic acid decarboxylase. Abbreviations: A, ApaI; B, BamHI; Bx, BstXI; E, EcoRI; H, HindIII; N, NotI; P, PstI; S, SalI; Sc, SacI; X, XhoI.