There is a recent and growing interest in the study and application of non‐Saccharomyces yeasts, mainly in fermented foods. Numerous publications and patents show the importance of these yeasts. However, a fundamental issue in studying and applying them is to ensure an appropriate preservation scheme that allows to the non‐Saccharomyces yeasts conserve their characteristics and fermentative capabilities by long periods of time. The main objective of this review is to present and analyze the techniques available to preserve these yeasts (by conventional and non‐conventional methods), in small or large quantities for laboratory or industrial applications, respectively. Wine fermentation is one of the few industrial applications of non‐Saccharomyces yeasts, but the preservation stage has been a major obstacle to achieve a wider application of these yeasts. This review considers the preservation techniques, and clearly defines parameters such as culturability, viability, vitality and robustness. Several conservation strategies published in research articles as well as patents are analyzed, and the advantages and disadvantages of each technique used are discussed. Another important issue during conservation processes is the stress to which yeasts are subjected at the time of preservation (mainly oxidative stress). There is little published information on the subject for non‐Saccharomyces yeast, but it is a fundamental point to consider when designing a preservation strategy.
Alcoholic fermentation is influenced by yeast strain, culture media, substrate concentration and fermentation conditions, which contribute to taste and aroma. Some non-Saccharomyces yeasts are recognized as volatile compound producers that enrich aromatic profile of alcoholic beverages. In this work, 21 strains of Pichia kluyveri isolated from different fermentative processes and regions were evaluated. A principal component analysis (PCA) showed statistical differences between strains mainly associated with the variety and concentration of the compounds produced. From the PCA, two strains (PK1 and PK8) with the best volatile compound production were selected to evaluate the impact of culture media (M12 medium and Agave tequilana juice), stirring speeds (100 and 250 rpm) and temperatures (20°C, 25°C and 30°C). Increased ester production was observed at 250 rpm. Greatest effect in alcohols and ester production was found with A. tequilana, identifying PK1 as higher alcohol producer, and PK8 as better ester producer. Regarding temperature, PK1 increased ester production with decreased fermentation temperature. PK8 presented maximum levels of ethyl acetate and ethyl dodecanoate at 20°C, and finally isoamyl acetate increased its production at 30°C. Therefore, P. kluyveri strains are of great interest to produce different aromatic profiles that are affected by factors including medium, agitation and temperature.
Agro-industrial wastes can be used to obtain high-value compounds rich in antioxidant and antimicrobial activity. This study aimed to compare different extraction techniques for the recovery of sugars, antioxidants, and antimicrobial compounds from brewer’s spent grain (BSG), blue agave bagasse (BAB), spoiled blackberries (BB), and raspberries (RB). Aqueous (AQ), enzymatic (E), chemical-enzymatic (CE), and hydroalcoholic (EOH) extractions were assessed, and sugars, phenolics, flavonoids, and anthocyanin contents were quantified. Antioxidant activity of the extracts was evaluated using the ABTS and DPPH assays, and antimicrobial activity was tested against three yeasts and six bacteria. The CE process gave the highest total and reducing sugars content for the four residues tested, and the highest antioxidant activity, phenolics, flavonoids and anthocyanin content for BAB and BSG. Regarding BB and RB, the best treatment to obtain total and reducing sugars and antioxidant activity with ABTS was CE; the highest content of anthocyanins, phenolic, flavonoids and antioxidant activity with DPPH was obtained with EOH treatment. CE extracts of BSG and RB showed the highest inhibition against the strains studied. Results show that BSG, BB, and RB can be a source of antioxidants and antimicrobial compounds for the food and pharmaceutical industries. Depending on the desired application and component of interest, one of the extraction techniques evaluated here could be used.
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