Spores and hyphal fragments of S. l

Spores and hyphal fragments of S. lividans transformants grown on A9 (with 100 mg apramycin l21) were inoculated into 250 ml baffled flasks containing 40 ml A345 and shakenat 240 r.p.m. for 44 h at 30 uC. Five per cent of this seed was transferred to 250 ml baffled flasks containing 50 ml A346 and shaken at240 r.p.m. for up to 7 days at 30 uC. For production in fermenters,biomass from 9-day cultures of S. lividans CBUK 136742 on A9 wasinoculated into a 2 l baffled Erlenmeyer flask containing 250 mlA355 (with 0?1 % antifoam A, 25 mg apramycin l21) and shaken at240 r.p.m. for 2 days at 30 uC. This seed was used to inoculate14 l A346 (with 0?1 % antifoam A) in a 20 l fermenter stirred at350 r.p.m., aerated at 0?7 v.v.m. and maintained at 30 uC. After20 h, a 50 % (w/v) glucose feed at 5 g h21 was introduced. After40 h, a 50 : 50 (w/w) blend of decanoic acid : methyl oleate feed at0?5gh21 was initiated.