Dialysis experiments show that physical interaction of TKS and OAC is not required for OA formation. Recombinant TKS and OAC were assayed in dialysis chambers separated by a 5-kDa cutoff membrane. (A) Assays with TKS and OAC in separate chambers resulted in the formation of HTAL, PDAL, and olivetol in the TKS-containing chamber 1 and HTAL, PDAL, and OA in the OAC-containing chamber 2. (B) Positive control assays with TKS and OAC in chamber 1 and no enzyme in chamber 2 produced large amounts of OA, in addition to HTAL and PDAL. (C) Negative control assays with TKS in chamber 1 and no enzyme in chamber 2 yielded only HTAL, PDAL, and olivetol. Chromatograms were extracted at 270 nm.