Influence of pulp on the microbial diversity during cupuassu fermentation

Cupuassu (Theobroma grandiflorum Schum) is a fruit belonging to the same genus as cocoa and, through seed fermentation, a chocolate-like product called "the cupulate" is obtained. The pulp is removed from the seeds before fermentation because its abundance hinders the process. Unlike cocoa, little is known about the microbial diversity involved in cupuassu fermentation. The goal of this study was to explore the use of next-generation sequencing to identify the yeasts and bacteria communities involved in cupuassu seed fermentation on three different pulp concentrations (0, 7.5, and 15%) as well as two turning schemes on the microbial growth. In order to do that, a massive sequencing of the 16S and ITS4 rRNA region (S) using the Illumina MiSeq Platform identified some genera of bacteria and yeasts, respectively, in the fermentation environment. Taxonomic analyses of both communities, especially at the genus level, revealed a predominance of yeasts such as Pichia and Hanseniaspora, and bacteria such as Acetobacter and Lactobacillus. A predominance of bacteria over yeasts diversity was observed in the experiments with higher pulp concentrations (15%). The physicochemical analysis showed that fermentation of samples with 15% pulp exhibited longer fermentation times, the highest temperatures, and elevated production of organic acids such as acetic acid, a precursor of flavor. In addition, the turning applied every 24 h to the mass slightly favored the formation of flavor precursors. It seems that the abundance and composition of cupuassu pulp, rich in organic compounds, can influence the diversity of some populations of yeasts. Some of those compounds identified in previous studies are terpenes with antimicrobial activity. More studies will be necessary to confirm if the presence of terpenes compounds in the cupuassu pulp exert some inhibitory action on microorganism diversity.