Microbial community structure, physicochemical characteristics and predictive functionalities of the Mexican tepache fermented beverage

Abstract

Tepache is a native beverage from Mexico, which is usually elaborated with pineapple shells, brown cane sugar and is fermented naturally. Beneficial health effects have been attributed to its consumption; however, the total ecosystem of this beverage including chemicals (substrates for microbial growth, prebiotics, etc) and microbiota (probiotics), and potential functionality had not been studied. In this work, the analysis of the tepache beverage for its physicochemical characteristics, as well as its structure of microbial communities and the predictive metabolic functionalities was carried out. Chemical characterization was performed via enzymatic and GC-MS methods. The bacterial and fungal communities were identified by using 16S rRNA and ITS metabarcoding through Illumina MiSeq 2 × 300. The metabolic potential was predicted by in silico tools. This research showed that after 72 h of fermentation, the tepache physicochemical characteristics shifted to 9.5 Brix degrees and acidic pH. The content of ethanol, acetic and L-lactic acid increased significantly from 0.83 ± 0.02 to 3.39 ± 0.18 g/L, from 0.38 ± 0.04 to 0.54 ± 0.04 g/L and from 1.42 ± 0.75 to 8.77 ± 0.34 g/L, respectively. While, the total sugars was decreased from 123.43 ± 2.01 to 84.70 ± 2.34 g/L. The microbial diversity analysis showed a higher richness of bacterial communities and increased fungal evenness at the end of fermentation. At 72 h of fermentation the microbial community was dominated by Lactobacillus, Leuconostoc, Acetobacter and Lactococcus bacterial genera. As for the fungal community, Saccharomyces, Gibberella, Zygosaccharomyces, Candida, Meyerozyma, Talaromyces, Epicoccum and Kabatiella were found to be in most abundance. The predicted functionality profile evidenced a close-fitting relationship between fungal communities at 0 h with the bacterial communities at 72 h of fermentation. The metabolic potential showed that glycolysis and citrate cycle metabolism were predominant for fungal community, while glycolysis, fructose and tricarboxylic acid metabolism were more representative for the bacterial core. Tepache fermentation mainly occurred at two temporal successions. First, a lactic acid and ethanol fermentation dominated by lactic acid bacteria and yeast, and then an increase in acetogenic bacteria. This study revealed for the first time the physicochemical, microbiological changes and predictive functionality that are involved during tepache fermentation. These findings contributed to the knowledge of important microbial sources and could be essential to future efforts in manufacturing process. In addition, this work could help to analyze the health benefits that are empirically attributed to it by consumers.

Keywords: 16S rRNA gene; Bacteria; Fungi; ITS gene; Illumina-MiSeq; Lactic acid.