*Article* **Kombucha Tea as a Reservoir of Cellulose Producing Bacteria: Assessing Diversity among** *Komagataeibacter* **Isolates**

**Salvatore La China, Luciana De Vero, Kavitha Anguluri, Marcello Brugnoli, Dhouha Mamlouk and Maria Gullo \***

Department of Life Sciences, University of Modena and Reggio Emilia, 42122 Reggio Emilia, Italy; salvatore.lachina@unimore.it (S.L.C.); luciana.devero@unimore.it (L.D.V.); kavitha.anguluri@unimore.it (K.A.); marcello.brugnoli96@gmail.com (M.B.); mamloukdhouha@gmail.com (D.M.) **\*** Correspondence: maria.gullo@unimore.it

**Abstract:** Bacterial cellulose (BC) is receiving a great deal of attention due to its unique properties such as high purity, water retention capacity, high mechanical strength, and biocompatibility. However, the production of BC has been limited because of the associated high costs and low productivity. In light of this, the isolation of new BC producing bacteria and the selection of highly productive strains has become a prominent issue. Kombucha tea is a fermented beverage in which the bacteria fraction of the microbial community is composed mostly of strains belonging to the genus *Komagataeibacter*. In this study, Kombucha tea production trials were performed starting from a previous batch, and bacterial isolation was conducted along cultivation time. From the whole microbial pool, 46 isolates were tested for their ability to produce BC. The obtained BC yield ranged from 0.59 g/L, for the isolate K2G36, to 23 g/L for K2G30—which used as the reference strain. The genetic intraspecific diversity of the 46 isolates was investigated using two repetitive-sequence-based PCR typing methods: the enterobacterial repetitive intergenic consensus (ERIC) elements and the (GTG)5 sequences, respectively. The results obtained using the two different approaches revealed the suitability of the fingerprint techniques, showing a discrimination power, calculated as the D index, of 0.94 for (GTG)5 rep-PCR and 0.95 for ERIC rep-PCR. In order to improve the sensitivity of the applied method, a combined model for the two genotyping experiments was performed, allowing for the ability to discriminate among strains.

**Keywords:** kombucha tea; microbial diversity; bacterial cellulose; *Komagataeibacter xylinus*; repetitive elements sequence-based rep-PCR; typing
