General guidelines on how to grow up a culture of Acetobacter xylinum, ATCC strain 53582 .
To prepare ～500 ml of liquid Acetobacter media, add the following:
Glucose - 1.0 g
Peptone - 2.5 g
Yeast extract - 2.5 g
Na2HPO4 - 1.35 g
Citric acid - 0.75 g
Distilled water - 500 ml
If you are making plates, use the same protocol but add 7.5 g of agar.
Prepare media as outlined
Autoclave to sterilize media.
Streak/inoculate Acetobacter onto plates or in media.
Incubate cells at 26°C for 2-3 days.
If using a freeze dried source of Acetobacter (ex. ATCC shipment), growth may take up to 4 days.
All questions, input and feedback are welcome!
The growth of Acetobacter does not give a cloudy appearance in the media, the media will remain transparent to slightly translucent in appearance.
The growth of Acetobacter is accompanied by the formation of a thick cellulose matrix within the media that must be removed before cells can be pelleted for a miniprep procedure. Simply vortex briefly to break up the cellulose into chunks and remove the cellulose chunks from the media with a pipette while carefully avoiding the removal of cells.
Acetobacter will grow well at room temperature in aerobic conditions.
For information on the growth conditions of other Acetobacter strains, please visit