867
867 . RHODOBACTER VELDKAMPII MEDIUM

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Culture medium recipe

Vol.

×

Main sol. 867 #

ml

Equipment needed: Autoclave

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
Medium 28	1000.00	ml	-	-
Yeast extract	0.25	%	2.5	-
Na-acetate	0.30	%	3	36.57
Vitamin B₁₂	2.00	mg	0.002	0.001
Standard vitamin solution	10.00	ml	-	-
1 supplement medium 28 with 0,25% Yeast-extract, 0,3% Na-acetate, 2mg/l Vitamin B₁₂ and 10ml/l Standard vitamin solution (see below).

Medium 28 #

1000

Solution occurs in 8 other media (first defined in Medium 28).

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
Solution A	460	ml	-	-
1 Aliquot Solution A into 100 mL screw-cap bottles, filled with 46 mL each. Bubble with N₂/CO₂ and autoclave at 121°C for 15 min (as decribed below).
2 Prepare the following solutions (resazurin, bicarbonate and Pfennig's heterotrophic salts) and sterilize as given below.
Resazurin solution	450	ml	-	-
Bicarbonate solution	50	ml	-	-
Pfennig's heterotrophic salts solution	26	ml	-	-
3 Add bicarbonate solution and Pfennig's heterotrophic salts to the resazurin (complete volumina, i.e. 50 mL bicarbonate solution and 26 mL Pfennig's heterotrophic salts solution). Bubble with CO₂ in an ice bath under sterile conditions.
4 Fill 50 ml of this mixture to each bottle of solution A (46 mL + 50 mL).
5 Before use, add 4 ml sulfide solution (1.5%) and 0.1 ml Vitamin B₁₂ solution to each 100 mL bottle.
Sulfide solution, 1.5%	40	ml	-	-
Vitamin B12 solution	1	ml	-	-
6 Adjust the pH with filter-sterilised 1M Na₂CO₃ to 7.1-7.3.
7 If needed, aliquot into sterile, N₂ gassed screw-cap tubes under N₂ gas.
8 During the first 24 h, the iron of the medium precipitates in the form of black flocks. No other sediment should arise in the otherwise clear medium.
9 Feed the actively growing culture periodically with neutralized 3% solution of sodium sulfide (use 1 -3 mL/100 mL depending on strain and cultivation stage) to replenish sulfide and with other supplement solutions (see Ref. 3365).
Neutralized sulfide solution 3% (w/v)	10	ml	-	-

Solution A #

460

Solution occurs in 4 other media (first defined in Medium 28).

Equipment needed: Autoclave

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
CaCl₂ x 2 H₂O	0.25	g	0.543	3.697
Yeast extract	0.25	g	0.543	-
Distilled water	460.00	ml	-	-
1 Aliquot Solution A into 100 mL screw-cap bottles, filled with 46 mL each. Bubble with N₂/CO₂ and autoclave at 121°C for 15 min.

Sulfide solution, 1.5% #

135

Solution occurs in 4 other media (first defined in Medium 28).

Equipment needed: Autoclave

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
Na₂S x 9 H₂O	2	g	14.815	61.682
Distilled water	135	ml	-	-
1 Prepare in a screw-cap bottle, bubble with N₂ to replace air, close tightly and autoclave.

Bicarbonate solution #

50

Solution occurs in 4 other media (first defined in Medium 28).

Equipment needed: Filter

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
NaHCO₃	1.5	g	30	357.113
H₂O	50.0	ml	-	-
1 Bubble with CO₂ and filter sterilize into sterile, gas-tight, 100 ml screw-cap bottle.

Resazurin solution #

450

Solution occurs in 4 other media (first defined in Medium 28).

Equipment needed: Autoclave Ice bath

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
Resazurin (0,1%)	0.5	ml	0.005	0.022
Distilled water	450.0	ml	-	-
1 Autoclave in a cotton-stoppered Erlenmeyer flask with an outlet tube for medium, connected to a glass outlet at the bottom of the vessel and has, at the other end, a silicon rubber tube with a pinch cock and a bell for aseptic dispensing of the medium into bottles.
2 Cool to room temperature under an atmosphere of N₂/CO₂ in an ice bath.

Pfennig's heterotrophic salts solution #

26

Solution occurs in 4 other media (first defined in Medium 28).

Equipment needed: Filter

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
Ammonium chloride	0.35	g	13.462	251.665
Ammonium acetate	0.25	g	9.615	124.746
Pyruvic acid sodium salt	0.25	g	9.615	87.381
Dextrose	0.25	g	9.615	48.521
MgSO₄ x 7 H₂O	0.50	g	19.231	78.023
KCl	0.35	g	13.462	180.571
KH₂PO₄	0.35	g	13.462	98.92
Trace element solution SL-12 B	1.00	ml	-	-
Distilled water	25.00	ml	-	-
1 Filter sterilize into sterile, gas-tight, 100 ml screw-cap bottle.

Vitamin B12 solution #

100

Solution occurs in 5 other media (first defined in Medium 28).

Equipment needed: Filter

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
Vitamin B₁₂	0.01	g	0.1	0.074
Distilled water	100.00	ml	-	-
1 Filter sterilized

Standard vitamin solution #

100

Solution occurs in 3 other media (first defined in Medium 81).

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
Riboflavin	10.0	mg	0.1	0.266
Thiamine-HCl x 2 H₂O	50.0	mg	0.5	1.339
Nicotinic acid	50.0	mg	0.5	4.061
Pyridoxine hydrochloride	50.0	mg	0.5	2.431
Calcium pantothenate	50.0	mg	0.5	1.049
Biotin	0.1	mg	1.0e-3	0.004
Folic acid	0.2	mg	0.002	0.005
Vitamin B₁₂	1.0	mg	0.01	0.007
Distilled water	100.0	ml	-	-

Neutralized sulfide solution 3% (w/v) #

100

Solution occurs in 18 other media (first defined in Medium 28).

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
Na₂S x 9 H₂O	3	g	30	124.905
Distilled water	100	ml	-	-
1 The sulfide solution is prepared in a 250 ml screw-capped bottle with a butyl rubber septum and a magnetic stirrer. The solution is bubbled with nitrogen gas, closed and autoclaved for 15 min. at 121°C. After cooling to room temperature the pH is adjusted to about 7.0 by adding of sterile 2 M H₂SO₄ drop-wise with a syringe without opening the bottle.

Metadata

Last modified:	24.02.22
Source:	DSMZ
Medium type:	Complex medium
Final pH:	n.d.
Equipment needed:	Autoclave, Filter, Ice bath

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Associated strains

All strains for this medium:

No strains found

Compound	Conc. [g/L]	Conc. [mM]
Na-acetate	3	36.57
NaHCO₃	1.5	17.856
Na₂S x 9 H₂O	0.593	2.467
MgSO₄ x 7 H₂O	0.5	2.029
Ammonium chloride	0.35	6.543
KCl	0.35	4.695
KH₂PO₄	0.35	2.572
Ammonium acetate	0.25	3.243
Pyruvic acid sodium salt	0.25	2.272
CaCl₂ x 2 H₂O	0.25	1.701
Dextrose	0.25	1.262
Yeast extract	0.25	-
Nicotinic acid	0.005	0.041
Pyridoxine hydrochloride	0.005	0.024
Thiamine-HCl x 2 H₂O	0.005	0.013
Calcium pantothenate	0.005	0.01
Na₂-EDTA	0.003	0.009
Resazurin	0.002	0.01
FeSO₄ x 7 H₂O	0.001	0.004
Riboflavin	1.0e-3	0.003
Vitamin B₁₂	1.0e-3	7.4e-4
H₃BO₃	3.0e-4	0.005
CoCl₂ x 6 H₂O	1.9e-4	0.001
MnCl₂ x 2 H₂O	5.0e-5	-
ZnCl₂	4.2e-5	3.1e-4
NiCl₂ x 6 H₂O	2.4e-5	1.9e-4
Folic acid	2.0e-5	4.5e-5
Na₂MoO₄ x 2 H₂O	1.8e-5	7.4e-5
Biotin	1.0e-5	4.1e-5
CuCl₂ x 2 H₂O	2.0e-6	1.2e-5
H₂O	-	-
Distilled water	-	-
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