- 10^7 Spores ofS. coelicolorM1152 [18]orS. coelicolorM145
[12]. - Sterile water.
- Razor blades.
- Ethyl acetate, HPLC grade (seeNote 1).
- Methanol, HPLC grade.
- 30C Incubator.
- Round-bottom flask.
- Rotary evaporator.
- 250-ml conical flasks.
- 500-ml conical flask.
- 2-ml plastic tubes.
- Speed vacuum concentrator.
- Vortex.
2.2 Extraction
of Butyrolactones
from Liquid Culture
- Supplemented minimal medium [17]: 5% (w/v) PEG 6000,
0.5 mM MgSO 4 ·7H 2 O, 25 mM TES buffer at pH 7.2, 0.5 mM
NaH 2 PO 4 , 0.5 mM K 2 HPO 4 , 50 mM glucose, 0.01% (w/v)
antifoam 289, 0.1% (v/v) trace element solution (Subheading
2.1,item 1), 0.2% (w/v) casamino acids (Difco). Autoclave all
components individually before mixing, except the glucose and
the trace elements, which are filter-sterilized before mixing. - 10^10 Spores of S. coelicolor M1152 [18]orS. coelicolor
M145 [12]. - 250-ml conical flask with stainless steel spring.
- 500-ml conical flask.
- Ethyl acetate, HPLC grade (seeNote 1).
- Methanol, HPLC grade.
- 30C shaking incubator.
- Round-bottom flask.
- Rotary evaporator.
- Separation funnel.
- Spectrophotometer.
- Disposable 1-ml cuvettes.
- 50-ml plastic tubes.
- Centrifuge.
- 2-ml plastic tubes.
- Vortex.
- MgSO 4 (anhydrous).
- Filter paper.
120 Marc Biarnes-Carrera et al.