To generate this extract, the strain was revived from freezer stock onto TSA and incubated at 37°C overnight.
A single colony was used to inoculate a 4mL culture of TSB, which was incubated at 37°C in a drum roller overnight.
25uL of overnight culture was used to inoculate 25mL of TSB in a 125mL Ultra Yield flask, which was grown at 37°C at 220rpm for 24hrs. 

The culture was centrifuged @3000rpm for 15 minutes to pellet cells.
Supernatant was decanted and acidified to 0.1% formic acid.
The acidified sample was centrifuged @3000rpm for 15 minutes to remove precipitates before being loaded onto a 60mg HLB SPE column. 

Once loaded, the SPE column was washed with 4mL H2O and 4mL of 10% ACN before elution with 3mL of 80% ACN. 

Elution was rotovapped to remove the organic and lyophilized to dryness. 

The sample was then rehydrated in 5% ACN w/ 0.1% FA to ~2mg/mL. 

2uL were injected into a Synergi Fusion-RP column with a flow rate of 200uL/min. 
	A: H2O w/ 0.1% FA
	B: ACN w/ 0.1% FA
Gradient: 5% B --> 60% B at 35min --> 98% B at 54 min

Scan Range: 250-3750m/z
Scan Rate ~6/second
Top 5 most intense ions --> 25eV collision energy for HCD