A procedure for designing starter cultures for fermentation is illustrated for prickly pear wine production. The illustration includes kinetic studies on inoculated and spontaneous fermentation, microorganism identification studies based on molecular biology tools, and microbial ecology studies, which led to the selection of strains that are capable of synthesizing alcohol and desirable volatile compounds. Results show that a mixed starter inoculum containing Pichia fermentans and Saccharomyces cerevisiae leads to a fermented product that contains 8.37% alcohol (v/v). The gas chromatography and mass spectrometry (GC-MS) analysis shows the presence of 9 major volatile compounds (Isobutanol, Isopentanol, Ethyl acetate, Isoamyl acetate, Ethyl octanoate, Ethyl decanoate, Ethyl 9-decanoate, β-Phenylethyl acetate, and Phenylethyl alcohol) that have ethereal, fruity, aromatic notes that are considered to be essential for a fine wine flavor. These compounds harmonically synergize with the alcohol to produce a fermented product with a unique flavor and taste. Several assays using the mixed culture show that the process is stable, predictable, controllable, and reproducible. Moreover, the results show that a mixed culture leads to a broader range of aromatic products than that produced by a single, pure culture. Therefore, we conclude that combinations of Saccharomyces strains and non-Saccharomyces strains can be used to obtain high-quality fermented beverages from prickly pear juice.

This paper describes a methodology to establish an optimal process design for prickly pear wine production that preserves the peculiar and unique traits of traditional products, generating at the same time, technical information for appropriate design of both bioreactor and overall process. The strategy includes alcoholic fermentation optimization by the mixed native culture composed by Pichia fermentans and Saccharomyces cerevisiae, followed by malolactic fermentation optimization by Oenococcus oeni. The optimization criteria were based on multiple output functions: alcohol content, volatile compounds profile, organic acids profile, and compound contents related to color, which were analyzed by spectroscopy-chromatography methods and sensory analysis. The results showed that the mixed culture inoculated into a bioreactor containing prickly pear juice with 20 °Bx of fermentable sugars concentration, processed at a constant temperature of 20 °C for 240 h, leads to a fermented product with 9.93% (v/v) total alcohol content, and significant abundance of volatile compounds, which provide fruity and ethereal aromatic notes, complemented by a lively but not unpleasant acidity. This young wine was further subjected to malolactic fermentation at constant temperature (16 °C) for 192 h, decreasing malic acid, and balancing volatile compounds contents, thus resulting in a product with better aroma and flavor perception, and a velvety feeling of long aftertaste. Repeated assays showed that the process is stable, predictable, controllable, and reproducible. These results were used for process design and spreadsheet construction in order to simulate the process, and properly select and size the equipment required for such process.