International Journal of Innovative Approaches in Agricultural Research
Abbreviation: IJIAAR | ISSN (Online): 2602-4772 | DOI: 10.29329/ijiaar

Research article    |    Open Access
International Journal of Innovative Approaches in Agricultural Research Volume 10 (2026)

Application of Metschnikowia Pulcherrima in Sequential Fermentations for the Production of Pilsner-Style Craft Beer

Julian Karaulli, Nertil Xhaferaj, Bruno Testa, Francesco Letizia, Onejda Kycyk, Mamica Ruci, Renata Kongoli, Fatbardha Lamce, Ilir Lloha, Kapllan Sulaj, Massimo Iorizzo

pp. 1 - 14   |  DOI: https://doi.org/10.29329/ijiaar.2026.1423.9

Publish Date: May 26, 2026  |   Single/Total View: 0/0   |   Single/Total Download: 0/0

PDF Download

Abstract

Previous studies have demonstrated promising potential and interest in the application of Metschnikowia pulcherrima in sequential fermentations in co-culture with Saccharomyces cerevisiae for the production of ale-style craft beer. This technique has yielded favorable results for top-fermenting beers, while its application in lager-style fermentations remains more limited. For this purpose, the indigenous S. cerevisiae 31 and M. pulcherrima 62 strains, isolated respectively from the grape varieties Shesh i Zi and Kallmet, were used to ferment a Pilsner-style beer in sequential inoculation. As a reference, a fermentation was prepared using the commercial lager strain S. pastorianus S-23 as a single starter. Fermentation kinetics and key physicochemical parameters were evaluated. The results showed that co-inoculation with M. pulcherrima 62 facilitated the completion of fermentation of S. cerevisiae 31, unlike the fermentation carried out with S. cerevisiae 31 alone. Furthermore, the co-cultures exhibited similar final alcohol content and pH values to those observed with the commercial strain S-23. However, higher levels of acetaldehyde were detected in beer obtained by sequential inoculation, while, glycerol concentrations were lower compared to beer obtained with S. pastorianus S-23. These preliminary findings suggest that the use of local non-Saccharomyces yeasts in Pilsner-style bottom fermentations is technologically feasible. Although not yet equivalent to commercial strains in performance, these findings are promising and provide a basis for the development of novel beer profiles using locally isolated starter cultures.

Keywords: Saccharomyces Cerevisiae, Metschnikowia Pulcherrima, Fermentation Characteristics, Sequential Inoculation, Pilsner Style, Craft Beer

How to Cite this Article?

APA 7th edition
Karaulli, J., Xhaferaj, N., Testa, B., Letizia, F., Kycyk, O., Ruci, M., Kongoli, R., Lamce, F., Lloha, I., Sulaj, K., & Iorizzo, M. (2026). Application of Metschnikowia Pulcherrima in Sequential Fermentations for the Production of Pilsner-Style Craft Beer. International Journal of Innovative Approaches in Agricultural Research, 10(1), 1-14. https://doi.org/10.29329/ijiaar.2026.1423.9

Harvard
Karaulli, J., Xhaferaj, N., Testa, B., Letizia, F., Kycyk, O., Ruci, M., Kongoli, R., Lamce, F., Lloha, I., Sulaj, K. and Iorizzo, M. (2026). Application of Metschnikowia Pulcherrima in Sequential Fermentations for the Production of Pilsner-Style Craft Beer. International Journal of Innovative Approaches in Agricultural Research, 10(1), pp. 1-14.

Chicago 16th edition
Karaulli, Julian, Nertil Xhaferaj, Bruno Testa, Francesco Letizia, Onejda Kycyk, Mamica Ruci, Renata Kongoli, Fatbardha Lamce, Ilir Lloha, Kapllan Sulaj and Massimo Iorizzo (2026). "Application of Metschnikowia Pulcherrima in Sequential Fermentations for the Production of Pilsner-Style Craft Beer". International Journal of Innovative Approaches in Agricultural Research 10 (1):1-14. https://doi.org/10.29329/ijiaar.2026.1423.9
References
  1. Aquilani, B., Laureti, T., Poponi, S., & Secondi, L. (2015). Beer choice and consumption determinants when craft beers are tasted: An exploratory study of consumer preferences. Food Quality and Preference, 41, 214–224. https://doi.org/10.1016/j.foodqual.2014.12.005 [Google Scholar] [Crossref] 
  2. Bellut, K., & Arendt, E. K. (2019). Chance and challenge: Non-Saccharomyces yeasts in nonalcoholic and low alcohol beer brewing – A review. Journal of the American Society of Brewing Chemists, 77(2), 77–91. https://doi.org/10.1080/03610470.2019.1569452 [Google Scholar] [Crossref] 
  3. Benito, S., Hofmann, T., Laier, M., Lochbühler, B., Schüttler, A., Ebert, K., Fritsch, S., Röcker, J., & Rauhut, D. (2015). Effect on quality and composition of Riesling wines fermented by sequential inoculation with non-Saccharomyces and Saccharomyces cerevisiae. European Food Research and Technology, 241, 707–717. https://doi.org/10.1007/s00217-015-2497-y [Google Scholar] [Crossref] 
  4. Canonico, L., Agarbati, A., Galli, E., Comitini, F., & Ciani, M. (2023). Metschnikowia pulcherrima as biocontrol agent and wine aroma enhancer in combination with a native Saccharomyces cerevisiae. LWT - Food Science and Technology, 181, Article 114758. https://doi.org/10.1016/j.lwt.2023.114758 [Google Scholar] [Crossref] 
  5. Catarino, M., & Mendes, A. (2011). Non-alcoholic beer—A new industrial process. Separation and Purification Technology, 79(3), 342–351. https://doi.org/10.1016/j.seppur.2011.03.020 [Google Scholar] [Crossref] 
  6. Chorbadzhiev, P., Gerginova, D., & Simova, S. (2025). Weiss or Wit: Chemical profiling of wheat beers via NMR-based metabolomics. Foods, 14(9), Article 1621. https://doi.org/10.3390/foods14091621 [Google Scholar] [Crossref] 
  7. Comitini, F., Agarbati, A., Canonico, L., & Ciani, M. (2021). Yeast interactions and molecular mechanisms in wine fermentation: A comprehensive review. International Journal of Molecular Sciences, 22(14), Article 7754. https://doi.org/10.3390/ijms22147754 [Google Scholar] [Crossref] 
  8. Comitini, F., Gobbi, M., Domizio, P., Romani, C., Lencioni, L., Mannazzu, I., & Ciani, M. (2011). Selected non-Saccharomyces wine yeasts in controlled multistarter fermentations with Saccharomyces cerevisiae. Food Microbiology, 28(5), 873–882. https://doi.org/10.1016/j.fm.2010.12.001 [Google Scholar] [Crossref] 
  9. Drosou, F., Mamma, D., Tataridis, P., Dourtoglou, V., & Oreopoulou, V. (2022). Metschnikowia pulcherrima in mono or co-fermentations in brewing. BrewingScience, 75, 104–114. http://dx.doi.org/10.23763/BrSc22-09drosou [Google Scholar]
  10. Einfalt, D. (2021). Barley‑sorghum craft beer production with Saccharomyces cerevisiae, Torulaspora delbrueckii and Metschnikowia pulcherrima yeast strains. European Food Research and Technology, 247, 385–393. https://doi.org/10.1007/s00217-020-03632-7 [Google Scholar] [Crossref] 
  11. Eram, M. S., & Ma, K. (2013). Decarboxylation of pyruvate to acetaldehyde for ethanol production by hyperthermophiles. Biomolecules, 3(3), 578–596. https://doi.org/10.3390/biom3030578 [Google Scholar] [Crossref] 
  12. European Brewery Convention. (2007). Analytica - EBC. Hans Carl, Fachverlag. [Google Scholar]
  13. Fontana, M., & Buiatti, S. (2009). Amino acids in beer. In V. Preedy (Ed.), Beer in Health and Disease Prevention (pp. 273–284). Academic Press. [Google Scholar]
  14. Garavaglia, C., & Swinnen, J. (2017). The craft beer revolution: An international perspective. Choices, 32(3), 1–8. [Google Scholar]
  15. Han, X., Qin, Q., Li, C., Zhao, X., Song, F., An, M., Chen, Y., Wang, X., Huang, W., & Zhan, J. (2023). Application of non-Saccharomyces yeasts with high β-glucosidase activity to enhance terpene-related floral flavor in craft beer. Food Chemistry, 404, Article 134726. https://doi.org/10.1016/j.foodchem.2022.134726 [Google Scholar] [Crossref] 
  16. Ivit, N. N., Longo, R., & Kemp, B. (2020). The effect of non-Saccharomyces and Saccharomyces cerevisiae yeasts on ethanol and glycerol levels in wine. Fermentation, 6(3), Article 77. https://doi.org/10.3390/fermentation6030077 [Google Scholar] [Crossref] 
  17. Jolly, N. P., Varela, C., & Pretorius, I. S. (2014). Not your ordinary yeast: Non-Saccharomyces yeasts in wine production uncovered. FEMS Yeast Research, 14(2), 215–237. https://doi.org/10.1111/1567-1364.12111 [Google Scholar] [Crossref] 
  18. Karaoglan, S. Y., Jung, R., Gauthier, M., Kinćl, T., & Dostàlek, P. (2022). Maltose-negative yeast in non-alcoholic and low-alcoholic beer production. Fermentation, 8(6), Article 273. https://doi.org/10.3390/fermentation8060273 [Google Scholar] [Crossref] 
  19. Karaulli, J., Xhaferaj, N., Ruci, M., Testa, B., Letizia, F., Albanese, G., Kongoli, R., Lamçe, F., Kyçyk, O., Sulaj, K., & Iorizzo, M. (2023). Evaluation of Saccharomyces and non-Saccharomyces yeasts isolated from Albanian autochthonous grape varieties for craft beer production. In Proceedings of V International Agricultural, Biological, Life Science Conference (AGBIOL). Edirne, Turkey. [Google Scholar]
  20. Karaulli, J., Xhaferaj, N., Coppola, F., Testa, B., Letizia, F., Kyçyk, O., Kongoli, R., Ruci, M., Lamçe, F., Sulaj, K., & Iorizzo, M. (2024). Bioprospecting of Metschnikowia pulcherrima strains, isolated from a vineyard ecosystem, as novel starter cultures for craft beer production. Fermentation, 10(10), Article 513. https://doi.org/10.3390/fermentation10100513 [Google Scholar] [Crossref] 
  21. Karaulli, J., Xhaferaj, N., Testa, B., Letizia, F., Kyçyk, O., Ruci, M., Kongoli, R., Lamçe, F., Lloha, I., Sulaj, K., & Iorizzo, M. (2024). Application of Saccharomyces cerevisiae 31 and Metschnikowia pulcherrima 62, isolated from Albanian vineyards, as new starters in the production of Ale style beer [Conference presentation]. International Biological & Life Sciences Congress, Antalya, Turkey. [Google Scholar]
  22. Klimczak, K., Cioch-Skoneczny, M., Ciosek, A., & Poreda, A. (2024). Application of non-Saccharomyces yeast for the production of low-alcohol beer. Foods, 13(20), Article 3214. https://doi.org/10.3390/foods13203214 [Google Scholar] [Crossref] 
  23. Kregiel, D., Krajewska, A., Kowalska-Baron, A., Czarnecka-Chrebelska, K. H., & Nowak, A. (2024). Photoprotective effects of yeast pulcherrimin. Molecules, 29(20), Article 4873. https://doi.org/10.3390/molecules29204873 [Google Scholar] [Crossref] 
  24. Li, H., Han, X., Liu, F., Kun-Farkas, G., & Kiss, Z. (2015). Simple HPLC method for determining the glycerol content of beer. Journal of the American Society of Brewing Chemists, 73(4), 314–317. https://doi.org/10.1094/ASBCJ-2015-0814-01 [Google Scholar] [Crossref] 
  25. Lin, X., Tang, X., Han, X., He, X., Han, N., Ding, Y., & Sun, Y. (2022). Effect of Metschnikowia pulcherrima on Saccharomyces cerevisiae Pdh by-pass in mixed fermentation with varied sugar concentrations of synthetic grape juice and inoculation ratios. Fermentation, 8(9), Article 480. https://doi.org/10.3390/fermentation8090480 [Google Scholar] [Crossref] 
  26. Liu, C., Li, Q., Niu, C., Zheng, F., & Zhao, Y. (2018). Simultaneous determination of diethyl acetal and acetaldehyde during beer fermentation and storage process. Journal of the Science of Food and Agriculture, 98(12), 4733–4741. https://doi.org/10.1002/jsfa.9008 [Google Scholar] [Crossref] 
  27. MacGregor, A. W. (1996). Malting and brewing science: Challenges and opportunities. Journal of the Institute of Brewing, 102(2), 97–102. https://doi.org/10.1002/j.2050-0416.1996.tb00900.x [Google Scholar] [Crossref] 
  28. Mencher, A., Morales, P., Curiel, J. A., Gonzalez, R., & Tronchoni, J. (2021). Metschnikowia pulcherrima represses aerobic respiration in Saccharomyces cerevisiae suggesting a direct response to co-cultivation. Food Microbiology, 94, Article 103670. https://doi.org/10.1016/j.fm.2020.103670 [Google Scholar] [Crossref] 
  29. Merck KGaA. (2009). Manual of analysis methods for the brewery industry (Method 12 – Diacetyl). ASBC Method. [Google Scholar]
  30. Michel, M., Haslbeck, K., Ampenberger, F., Meier-Dörnberg, T., Stretz, D., Hutzler, M., Coelhan, M., Jacob, F., & Liu, Y. (2019). Screening of brewing yeast β-lyase activity and release of hop volatile thiols from precursors during fermentation. BrewingScience, 72, 179–186. [Google Scholar]
  31. Michel, M., Meier-Dörnberg, T., Jacob, F., Methner, F., Wagner, R. S., & Hutzler, M. (2016). Pure non-Saccharomyces starter cultures for beer fermentation with a focus on secondary metabolites and practical applications. Journal of the Institute of Brewing, 122(4), 569–587. https://doi.org/10.1002/jib.346 [Google Scholar] [Crossref] 
  32. Morata, A., Loira, I., Escott, C., del Fresno, J. M., Bañuelos, M. A., & Suárez-Lepe, J. A. (2019). Applications of Metschnikowia pulcherrima in wine biotechnology. Fermentation, 5(3), Article 63. https://doi.org/10.3390/fermentation5030063 [Google Scholar] [Crossref] 
  33. Moreira, M. T. G., Pereira, P. R., Aquino, A., Conte-Junior, C. A., & Paschoalin, V. M. F. (2022). Aldehyde accumulation in aged alcoholic beer: Addressing acetaldehyde impacts on upper aerodigestive tract cancer risks. International Journal of Molecular Sciences, 23(22), Article 14147. https://doi.org/10.3390/ijms232214147 [Google Scholar] [Crossref] 
  34. Oro, L., Ciani, M., & Comitini, F. (2014). Antimicrobial activity of Metschnikowia pulcherrima on wine yeasts. Journal of Applied Microbiology, 116(5), 1209–1217. https://doi.org/10.1111/jam.12446 [Google Scholar] [Crossref] 
  35. Postigo, V., O’Sullivan, T., Schuurman, T. E., & Arroyo, T. (2022). Non-conventional yeast: Behavior under pure culture, sequential and aeration conditions in beer fermentation. Foods, 11(22), Article 3717. https://doi.org/10.3390/foods11223717 [Google Scholar] [Crossref] 
  36. Sadoudi, M., Rousseaux, S., David, V., Alexandre, H., & Tourdot-Maréchal, R. (2017). Metschnikowia pulcherrima influences the expression of genes involved in PDH bypass and glyceropyruvic fermentation in Saccharomyces cerevisiae. Frontiers in Microbiology, 8, Article 1137. https://doi.org/10.3389/fmicb.2017.01137 [Google Scholar] [Crossref] 
  37. Snyman, L., Theron, L. W., & Divol, B. (2019). The expression, secretion and activity of the aspartic protease MpAPr1 in Metschnikowia pulcherrima IWBT Y1123. Journal of Industrial Microbiology and Biotechnology, 46(12), 1733–1743. https://doi.org/10.1007/s10295-019-02236-4 [Google Scholar] [Crossref] 
  38. Staub, C., Contiero, R., Bosshart, N., & Siegrist, M. (2022). You are what you drink: Stereotypes about consumers of alcoholic and non-alcoholic beer. Food Quality and Preference, 101, Article 104633. https://doi.org/10.1016/j.foodqual.2022.104633 [Google Scholar] [Crossref] 
  39. Stewart, G. G. (2017). The production of secondary metabolites with flavour potential during brewing and distilling wort fermentations. Fermentation, 3(4), Article 63. https://doi.org/10.3390/fermentation3040063 [Google Scholar] [Crossref] 
  40. Troianou, V., Toumpeki, C., Dorignac, E., Kogkou, C., Kallithraka, S., & Kotseridis, Y. (2019). Evaluation of Saccharomyces pastorianus impact to Sauvignon blanc chemical & sensory profile compared to different strains of S. cerevisiae/bayanus. BIO Web of Conferences, 12, Article 02025. https://doi.org/10.1051/bioconf/20191202025 [Google Scholar] [Crossref] 
  41. Vicente, J., Ruiz, J., Belda, I., Benito-Vàzquez, I., Marquina, D., Calderόn, F., Santos, A., & Benito, S. (2020). The genus Metschnikowia in enology. Microorganisms, 8(7), Article 1038. https://doi.org/10.3390/microorganisms8071038 [Google Scholar] [Crossref] 
  42. Vrînceanu, C. R., Diguță, F. C., Cudalbeanu, M. D., Ortan, A., Mihai, C., Bărbulescu, I. D., Frîncu, M., Begea, M., Matei, F., & Teodorescu, R. I. (2025). Exploring the potential of Torulaspora delbrueckii, Starmerella bacillaris, and Saccharomyces cerevisiae as a probiotic starter for craft beer production. Foods, 14(9), Article 1608. https://doi.org/10.3390/foods14091608 [Google Scholar] [Crossref] 
  43. Withers, E. T. (2017). The impact and implications of craft beer research: An interdisciplinary literature review. In Craft Beverages and Tourism (Vol. 1, pp. 11–24). Palgrave Macmillan. [Google Scholar]
  44. Zhao, X., Procopio, S., & Becker, T. (2015). Flavor impacts of glycerol in the processing of yeast fermented beverages: A review. Journal of Food Science and Technology, 52(12), 7588–7598. https://doi.org/10.1007/s13197-015-1977-y [Google Scholar] [Crossref] 
Meta
Volume 10 (2026)
Download
Metric
History
Related

Volume 10 Issue 1

January 2026

All Manuscript
Copyright © 2026 International Journal of Innovative Approaches in Agricultural Research.
Pen Academic Publishing
This work is licensed under a Creative Commons Attribution 4.0 International License. CC BY
All elements of this website have been created using the BOQ™ - eJournal Management System.
Pen Academic Publishing General Publication Policy: