Foods 2022, 11, 2029. https://doi.org/10.3390/foods11142029 www.mdpi.com/journal/foods Article Impact of Non-Saccharomyces Wine Yeast Strains on Improving Healthy Characteristics and the Sensory Profile of Beer in Sequential Fermentation Vanesa Postigo 1,2, *, Paula Sanz 1 , Margarita García 1 and Teresa Arroyo 1 1 Department of Agri-Food, Madrid Institute for Rural, Food and Agriculture Research and Development (IMIDRA), El Encín, A-2, km 38.2, 28805 Alcala de Henares, Spain; paula.sanz9991@gmail.com (P.S.); margarita_garcia_garcia@madrid.org (M.G.); teresa.arroyo@madrid.org (T.A.) 2 Brewery La Cibeles, Petróleo 34, 28918 Leganes, Spain * Correspondence: vanesa.postigo@madrid.org Abstract: The use of non-Saccharomyces yeasts in brewing is a useful tool for developing new prod- ucts to meet the growing consumer demand for innovative products. Non-Saccharomyces yeasts can be used both in single and in mixed fermentations with Saccharomyces cerevisiae, as they are able to improve the sensory profile of beers, and they can be used to obtain functional beers (with a low ethanol content and melatonin production). The aim of this study was to evaluate this capacity in eight non-Saccharomyces strains isolated from Madrid agriculture. For this purpose, single fermen- tations were carried out with non-Saccharomyces strains and sequential fermentations with non-Sac- charomyces and the commercial strain SafAle S-04. The Wickerhamomyces anomalus strain CLI 1028 was selected in pure culture for brewing beer with a low ethanol content (1.25% (v/v)) for its fruity and phenolic flavours and the absence of wort flavours. The best-evaluated strains in sequential fermentation were CLI 3 (Hanseniaspora vineae) and CLI 457 (Metschnikowia pulcherrima), due to their fruity notes as well as their superior bitterness, body, and balance. Volatile compounds and mela- tonin production were analysed by GC and HPLC, respectively. The beers were sensory-analysed by a trained panel. The results of the study show the potential of non-Saccharomyces strains in the production of low-alcohol beers, and as a flavour enhancement in sequential fermentation. Keywords: non-Saccharomyces; sequential fermentation; beer aroma; wine yeast 1. Introduction The brewing industry generally uses Saccharomyces yeasts for brewing, while non- Saccharomyces yeasts have been more characteristic of spontaneously fermenting beers such as Sour and Lambic beer styles. [1]. During fermentation, the sugars present in the wort are totally or partially trans- formed into secondary metabolites such as aromas (higher alcohols, esters, sulphur com- pounds, acids, vicinal diketones), CO2, ethanol, glycerol, and melatonin by the yeasts. Some compounds also affect the antioxidant capacity of the beer, thus determining the final quality of the beer [2–5]. Likewise, over the last few years, the craft beer industry has been growing and with it the interest on the part of consumers for new beers with different properties [6]. This demand has led to pure culture and sequential fermentations with non-conventional yeasts to obtain beers with a low ethanol content, as well as functional and good organo- leptic properties [7]. In this context, studies on wine are more abundant than on beer. Sequential or mixed fermentations in beer has been studied with species such as Pichia kluyveri, Brettanomyces spp., Torulaspora delbrueckii, Hanseniaspora guilliermondii, Han- seniaspora opuntiae, and Lachancea thermotolerans [8–12], and a production of esters and Citation: Postigo, V.; Sanz, P.; García, M.; Arroyo, T. Impact of Non-Saccharomyces Wine Yeast Strains to Improve the Sensory Profile of Beer in Sequential Fermentation. Foods 2022, 11, 2029. https://doi.org/10.3390/foods11142029 Academic Editors: Gabriella Siesto, Angela Capece, Rocchina Pietrafesa and Patrizia Romano Received: 12 June 2022 Accepted: 6 July 2022 Published: 8 July 2022 Publisher’s Note: MDPI stays neu- tral with regard to jurisdictional claims in published maps and institu- tional affiliations. Copyright: © 2022 by the authors. Li- censee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and con- ditions of the Creative Commons At- tribution (CC BY) license (http://crea- tivecommons.org/licenses/by/4.0/).