# 461b: MINERAL MEDIUM WITH PYRROLIC ACID


## Main sol. 461b

1000	ml	**Main sol. 461**	
10	ml	**Pyrrole-2-carboxylic acid solution**	

1. Prepare and autoclave medium 461 (vitamins not necessary). Add 10 ml of pyrrolic acid solution per liter mineral medium.


## Main sol. 461

1000	ml	Distilled water	
1.45	g	Na2HPO4 x 2 H2O	
0.25	g	KH2PO4	
0.01	g	CaCl2	
0.5	g	MgSO4 x 7 H2O	
0.01	g	MnSO4	
0.3	g	NH4Cl	
0.05	g	NaCl	
5	ml	**Vitamin solution**	
1	ml	**Trace element solution SL-10**	

1. Dissolve and autoclave the phosphates separately. Add the vitamin solution and trace element solution after autoclaving. After mixing, pH should be 7.5.
2. Rehydrate and cultivate the lyophilized cells in complex medium recommended for the specific strain (e.g. medium 1, 220 or 535) first. After this reactivation, cultivate on mineral medium 461 with the appropriate carbon source.


## Pyrrole-2-carboxylic acid solution

10	M	Pyrrole-2-carboxylic acid	
1000	ml	Potassium phosphate buffer (50 mM; pH 7.2)	

1. Dissolve 10 mol pyrrole-2-carboxylic acid per liter potassium phosphate buffer (50 mM; pH 7.2) and sterilize the solution by membrane filtration.