Cellulose production and cellulose synthase gene detection in acetic acid bacteria

• Published in: Applied microbiology and biotechnology 2015-02, Vol.99 (3), p.1349-1361
• Main Authors: Valera, Maria José, Torija, Maria Jesús, Mas, Albert, Mateo, Estibaliz
• Format: Article
• Language: English
• Subjects: Acetic acid; Acetic Acid - metabolism; acetic acid bacteria; Acetobacter; Amino Acid Sequence; Applied Genetics and Molecular Biotechnology; Bacteria; Bacteria - enzymology; Bacteria - genetics; Bacteria - isolation & purification; Bacteria - metabolism; Bacterial Proteins - chemistry; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; biofilm; Biofilms; Biomedical and Life Sciences; Biosynthesis; Biotechnology; Blueberry Plants - microbiology; Carbon; Carbon sources; Cellulose; Cellulose - biosynthesis; cellulose synthase; Ethanol; fluorescence microscopy; Gene expression; Genes; Genetic aspects; Gluconacetobacter; Gluconobacter; Glucose; Glucosyltransferases - chemistry; Glucosyltransferases - genetics; Glucosyltransferases - metabolism; Identification and classification; ISO standards; Laboratories; Life Sciences; Microbial Genetics and Genomics; Microbiology; Microscopy; Molecular Sequence Data; oxygen; Phylogeny; polymerase chain reaction; Proteobacteria; Sequence Alignment; Studies; Vinegar; vinegars; Vitis - microbiology
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/s00253-014-6198-1

The ability of acetic acid bacteria (AAB) to produce cellulose has gained much industrial interest due to the physical and chemical characteristics of bacterial cellulose. The production of cellulose occurs in the presence of oxygen and in a glucose-containing medium, but it can also occur during vinegar elaboration by the traditional method. The vinegar biofilm produced by AAB on the air-liquid interface is primarily composed of cellulose and maintains the cells in close contact with oxygen. In this study, we screened for the ability of AAB to produce cellulose using different carbon sources in the presence or absence of ethanol. The presence of cellulose in biofilms was confirmed using the fluorochrome Calcofluor by microscopy. Moreover, the process of biofilm formation was monitored under epifluorescence microscopy using the Live/Dead BacLight Kit. A total of 77 AAB strains belonging to 35 species of Acetobacter, Komagataeibacter, Gluconacetobacter, and Gluconobacter were analysed, and 30 strains were able to produce a cellulose biofilm in at least one condition. This cellulose production was correlated with the PCR amplification of the bcsA gene that encodes cellulose synthase. A total of eight degenerated primers were designed, resulting in one primer pair that was able to detect the presence of this gene in 27 AAB strains, 26 of which formed cellulose.