Acetobacter Xylinum Culture

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General guidelines on how to grow up a culture of Acetobacter xylinum, ATCC strain 53582 .

Preparation of Acetobacter Media

To prepare ∼500 ml of liquid Acetobacter media, add the following:

  • Glucose - 10 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.


  1. Prepare media as outlined
  2. Autoclave to sterilize media.
  3. Streak/inoculate Acetobacter onto plates or in media.
  4. Incubate cells at 26°C for 2-3 days.
  5. 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!

  1. The growth of Acetobacter does not give a cloudy appearance in the media, the media will remain transparent to slightly translucent in appearance.
  2. 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.
  3. Acetobacter will grow well at room temperature in aerobic conditions.
  4. For information on the growth conditions of other Acetobacter strains, please visit [1]



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