TABLE 1.
Production of AuAAC by different microorganisms
| Microorganism | AuAAC production (IU/ml)a at indicated time
|
||||
|---|---|---|---|---|---|
| 0 h | 72 h | 96 h | 120 h | 140 h | |
| A. utahensis NRRL 1205b | 0.060 | ||||
| S. lividans 1326b | 0 | 0 | 0 | 0 | 0 |
| S. lividans (pEM4)b | 0 | 0 | 0 | 0 | 0 |
| S. lividans (pEAAC)b | 0 | 0.695 | 1.264 | 0.849 | 0.861 |
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.