Objective: This study aimed for the isolation and identification of mutation -resistant cellulose producing acetic acid bacteria from fruits and organic vinegar samples through biochemical and molecular methods.

Materials and Methods: Strains that could produce acetic acid in calcium-carbonate (CaCO3)-ethanol culture media, and produce cellulose in Hestrin-Schramm (HS) culture media were identified in genus level by application of relevant biochemical tests. The amount of the cellulose produced was determined by cell count and glucose concentration analysis. Cellulose structure was verified by the micromethod described by Dearing and by XRD analysis. Most efficient strain, selected according to the high level of cellulose productivity and stability over long-term preservation, was identified at species level by 16S rRNA gene sequence analysis.

Results: A total of 112 colonies compatible with the colony morphologies of acetic acid bacteria were isolated. The majority (63.4%) were originated from the rotten fruits. It was determined that six of the 35 isolates that oxidised acetic acid, produced cellulose. Elaborate biochemical tests supported the affiliation of selected isolates to Gluocnacetobacter genus. Cellulose production capacities of these strains were determined within the range 0.56-4.7 g/l, after incubation in HS medium for a week at 30°. XRD analysis showed that only one strain was able to produce “Cellulose I crystal” type crystalline cellulose. However, acid hydrolysis of all samples confirmed cellulose structure due to recovery rates of 95.14%-98.57%. Only strain that could preserve its cellulose production capability over passages and long-term storage, was P2A isolated from a rotten plum. Comparison of the 16S rRNA sequence of this strain with similar sequences obtained from primary databanks revealed 99.8% sequence similarities with G. hansenii species and thus, the strain was named as G. hansenii P2A.

Conclusion: As a result, a strain that exhibited high production of bacterial cellulose (1.275±0.15 g/l) and resistance to long term preservation, was isolated and identified as G. hansenii P2A. The strain was included under the reference number KUEN 1606 in the culture collection of the “Microorganism Culture Collections Research and Application Center” Department of Microbiology Istanbul University Faculty of Medicine, Turkey. This strain was suggested as a candidate organism for the industrial production of bacterial cellulose.