Screening and characterizing indigenous yeasts, lactic acid bacteria, and acetic acid bacteria from cocoa fermentation in Hainan for aroma Development

Fermentation is crucial for inducing desirable flavor and aroma profiles in cocoa products. This research focused on identifying microbial strains isolated from spontaneous cocoa fermentation in Hainan through 16S and Internal Transcribed Spacer (ITS) sequencing. Pectinase activity was screened, and metabolic dynamics of sugars and organic acids were analyzed using high-performance liquid chromatography. Additionally, gas chromatography-mass spectrometry was employed for the quantification of volatile compounds. The fermentation potentials of isolated yeast, lactic acid bacteria, and acetic acid bacteria were analyzed from multiple perspectives. Pichia fermentans XY23.1 and Hanseniaspora uvarum XY23.1 exhibited significant pectinolytic activity, essential for breaking down pectin in cocoa pulp. Moreover, H. uvarum XY23.1, H. occidentalis XY23.1, Saccharomyces cerevisiae XY23.2, and P. fermentans XY23.1 were identified as producers of notable amounts of alcohols and esters, contributing sweet and floral notes to the fermentation profile. Furthermore, Levilactobacillus brevis exhibited strong fructophilicity, and Lactiplantibacillus plantarum strains showed high metabolic rates and lactic acid production abilities, crucial for enhancing fermentation efficiency. Assessment of growth rate and acid production performance revealed that Gluconobacter potus XY23.2 and Acetobacter oryzifermentans XY23.1 can produce less acid during rapid growth, avoiding flavor defects caused by excessive acidity. This study demonstrates the impact of various flavor compounds on the flavor characteristics of cocoa pulp. It highlights the potential of these microbial strains for use in starter culture cocktails, which can significantly improve the quality of cocoa products by enhancing desirable flavor and aroma profiles while maintaining balanced acidity. PRACTICAL APPLICATION: This study screened and characterized microorganisms isolated from the fermentation of Hainan cocoa (Trinitario) through a series of experiments, providing new insights for the future selection of cocoa fermentation starters.