TABLE 1.

Production of AuAAC by different microorganisms

MicroorganismAuAAC production (IU/ml)a at indicated time
0 h72 h96 h120 h140 h
A. utahensis NRRL 1205b0.060
S. lividans 1326b00000
S. lividans (pEM4)b00000
S. lividans (pEAAC)b00.6951.2640.8490.861
  • a IU, μmol of 6-APA produced per minute under the standard assay conditions. Acylase activity was routinely assayed using penicillin V as a substrate. The activities of A. utahensis NRRL 12502 and of the recombinant S. lividans culture supernatants were determined according to the method previously described (20).

  • b For enzymatic assays, A. utahensis was cultured in liquid medium containing 3% sucrose, 0.5% peptone, 0.1% K2HPO4, 0.05% KCl, 0.05% MgSO4·7H2O, and 0.0002% FeSO4·7H2O (pH 6.5) (18) at 30°C and 250 rpm for 96 h, whereas S. lividans was cultured aerobically under submerged conditions in trypticase soy broth with thiostrepton (5 μg/ml) at 30°C and 250 rpm.