Screening and molecular characterization of new thermo- and ethanol-tolerant Acetobacter malorum strains isolated from two biomes Moroccan cactus fruits

Biotechnol Appl Biochem. 2021 Jun;68(3):476-485. doi: 10.1002/bab.1941. Epub 2020 Jun 2.

Abstract

Industrially, the sensitivity of acetic acid bacteria (AAB) to the high temperatures and the high ethanol concentrations is the major concerns for manufacturers. This study was conceived and designed to isolate and identify new thermo- and ethanol-tolerant AAB from Opuntia ficus-indica L. fruits. As a result, among 140 isolated bacterial strains, five selected strains (CR1, CR5, CR23, CZ2, and CZ15) exhibited important acetic acid production until 40 °C. The use of 16S rDNA gene analysis was insufficient to identify selected bacteria. Indeed, except CR5 that presented 100% similarity to A. cerevisiae, the other strains presented similar homology rates simultaneously to the 16S rDNA sequences of A. cerevisiae and A. malorum. The reidentification by 16S-23S rDNA gene sequencing showed that CR1, CR23, and CZ15 were A. malorum, which were shown tolerance to the highest concentration of ethanol (12%) and produced elevated amount (40 g/L) of acetic acid at 37 °C. In summary, we showed the thermotolerance and ethanol tolerant character of new A. malorum strains, which can be used as a starter for vinegar production. Furthermore, during the molecular characterization of the isolated strains, we concluded that 16S-23S rDNA internal transcribed spacer sequence is of great importance for discriminating between AAB species as a complement to the identification by 16S rDNA sequencing.

Keywords: Acetobacter; acetic acid bacteria; cactus; fermentation; thermotolerance; vinegar.

MeSH terms

  • Acetobacter / genetics
  • Acetobacter / isolation & purification*
  • DNA, Bacterial / genetics
  • Ethanol / chemistry*
  • Fruit / microbiology*
  • Opuntia / microbiology*
  • RNA, Ribosomal, 16S / genetics
  • RNA, Ribosomal, 23S / genetics
  • Temperature*

Substances

  • DNA, Bacterial
  • RNA, Ribosomal, 16S
  • RNA, Ribosomal, 23S
  • Ethanol

Supplementary concepts

  • Acetobacter malorum