A novel strategy to construct multi-strain starter cultures: an insight to evolve from natural to directed fermentation

Some biotechnological strategies have succeeded in the attempt to imitate natural fermentation, and bioprocesses have been efficiently designed when the product is the result of a unique biological reaction. However, when the process requires more than one biological reaction, few bioprocesses have been successfully designed because the available tools to construct multi-strain starter cultures are not yet well defined. In this work, a novel experimental strategy to construct multi-strain starter cultures with selected native microorganisms from natural fermentation is proposed. The strategy analyses, selects, and defines the number and proportion of each strain that should form a starter culture to be used in directed fermentations. It was applied to evolve natural fermentation to directed fermentation in distilled agave production. The results showed that a starter culture integrated by Kluyveromyces marxianus, Clavispora lusitaniae, and Kluyveromyces marxianus var. drosophilarum in proportions of 35, 32, and 33%, respectively, allows obtaining fermented agave juice containing a 2.1% alcohol yield and a distilled product with a broad profile of aromatic compounds. Hence, the results show, for the first time, a tool that addresses the technical challenge for multi-strain starter culture construction, offering the possibility of preserving the typicity and genuineness of the original traditional product.