Modulation of cocoa fermentation by co-inoculation of Pichia kluyveri CCMA0237 and Hanseniaspora uvarum CCMA0236: targeted chemical profiling and chocolate quality.

This study evaluated the effects of co-inoculating Pichia kluyveri CCMA0237 and Hanseniaspora uvarum CCMA0236 during cocoa fermentation with the SJ02 hybrid on the chemical profiling and sensory quality of chocolate. Microbial dynamics were monitored using qPCR, chemical analyses by HPLC, LC/MS, and GC/MS, and sensory analysis using an acceptance test followed by CATA questions. The co-inoculated yeasts showed higher microbial density; H. uvarum reached the highest cell density, while P. kluyveri in the control was detected only at 72 h at a low concentration. Co-inoculation accelerated sugar metabolism, resulting in higher ethanol accumulation in the pulp. In the control, seeds showed higher acetic acid levels at 72 h. Methylxanthines showed higher levels in the control at the end of drying. Free amino acids increased at 144 h, especially in co-inoculation, with increases of 35.5% in total and 68.7% in essential ones. Chemical characterization revealed broader, more intense formation of key volatiles, particularly higher alcohols and ketones. Sensory evaluation supported these findings, with chocolates from co-inoculated fermentations showing stronger cocoa notes and reduced bitterness. These results demonstrate that targeted co-inoculation with P. kluyveri CCMA0237 and H. uvarum CCMA0236 enhances the chemical complexity and sensory quality of cocoa hybrid SJ02, offering a strategy to improve chocolate value.