Sour Beer Research Articles

Exploring the diversity of native Lachancea thermotolerans strains isolated by sugary extracts from manna ash to modulate the flavour of sour beers

Exploring the diversity of native Lachancea thermotolerans strains isolated by sugary extracts from manna ash to modulate the flavour of sour beers

Survival of the probiotic strain Lacticaseibacillus paracasei subsp. paracasei F19 in high-hopped beers

This study aims to enhance understanding of probiotic lactic acid bacteria (LAB) survival in high-hopped beer formulations and their interactions with different yeasts and highlights the fermentation processes, microbial metabolism, and production of distinctive beer flavors. For this, this research used Lacticaseibacillus paracasei F19 (F19), Saccharomycodes ludwigii, and Saccharomyces cerevisiae strains US-05 (US-05) and Kveik (Kveik) for brewing. Bacterial and yeast cultures were prepared, fermented in wort, and analyzed in different hop concentrations (International Bitterness Units − IBU 0, 20, 40). Methods included physicochemical analysis, yeast and bacterial counts, RT-qPCR for gene expression, statistical analysis, and sensory evaluation by sommeliers following BJCP guidelines. Physicochemical analysis showed efficient fermentation across all hop concentrations (IBU 0, 20, 40), with decreasing SG and pH over time due to lactic acid bacteria and yeast metabolism. Higher hop levels (IBU 20 and 40) resulted in less acidic beer, indicating hop interference with bacterial activity. Yeast populations remained stable regardless of hop content, with Saccharomyces cerevisiae and Saccharomycodes ludwigii performing well. Probiotic strain F19 exhibited robust viability in all formulations. Sensory analysis favored higher hop content beers, suggesting consumer acceptance and potential health benefits of probiotic, high-hop beers. Higher hop content hindered sour beer production as only hop-free beers reached low pH levels. Probiotic strain F19 remained viable under high IBU formulations (20 and 40), with these being preferred by sommeliers using BJCP methodology. All yeast strains supported F19 survival. Further studies are needed on gastrointestinal resistance and clinical benefits.

Chemical profiling and probiotic viability assessment in Gueuze-style beer: Fermentation dynamics, metabolite and sensory characterization, and in vitro digestion resistance

Probiotic viability, metabolite concentrations, physicochemical parameters, and volatile compounds were characterized in Gueuze beers formulated with probiotic lactic acid bacteria (LAB) and yeast. Additionally, the sensory profile of the beers and the resistance of the probiotics to digestion were determined. The use of 2 International Bitterness Units resulted in high concentrations of probiotic LAB but a decline in probiotic yeast as pH decreased. Secondary fermentation led to the consumption of maltose, citric acid, and malic acid, and the production of lactic and propionic acids. Carbonation and storage at 4 °C had minimal impact on probiotic viability. The addition of probiotic LAB resulted in a distinct aroma profile with improved sensory characteristics. Our results demonstrate that sour beers produced with probiotic LAB and a probiotic yeast, and fermented using a two-step fermentation process, exhibited optimal physicochemical parameters, discriminant volatile compound profiles, promising sensory characteristics, and high probiotic concentrations after digestion.

Lactic and Acetic Acids in Sour Beers Promote Corrosion During Aluminum Beverage Can Storage

Newer sour beers are often packaged in aluminum cans, but the compatibility of sour beer and its major acids (lactic, acetic) with cans is unclear. In an initial study, commercial sour beers were packaged in cans containing one of four different liners (bisphenol A (BPA) epoxy, two BPA–non-intent (BPA-NI) epoxy, and acrylic). Corrosion, as measured by dissolved aluminum and visual degradation of the liner, was positively correlated with concentrations of lactic acid, acetic acid, and decreased pH value. After 48 wk, aluminum concentrations up to 58 mg/L were observed in one sour beer, or nearly 100-fold greater than typical dissolved aluminum concentrations in non-sour beers. Liner type did not affect corrosion. In a subsequent model sour beer study with two acrylic liners and one BPA-NI liner, molecular SO2 positively correlated with corrosion, but only at concentrations 5-fold higher than the maximum expected in sour beers. Other added components (chloride, copper, ethanol) did not affect corrosion. Addition of acetic, lactic, and phosphoric acid in varying equinormal combinations to a non-sour beer demonstrated that acetic and lactic acids (average dissolved aluminum = 2.54 mg/L following storage) promote corrosion more than phosphoric acid (average dissolved aluminum = 0.47 mg/L). Titratable acidity (TA) correlated well with corrosion, with increased dissolved aluminum observed at TA > 6 g/L as lactic acid equivalents. Organic acid corrosivity was hypothesized to relate to the proportion of acid in its neutral form, and thus these findings are relevant to producers of other beverages with high levels of organic acids.

A Review on the Interaction of Acetic Acid Bacteria and Microbes in Food Fermentation: A Microbial Ecology Perspective.

In fermented foods, acetic acid bacteria (AAB), kinds of bacteria with a long history of utilization, contribute to safety, nutritional, and sensory properties primarily through acetic acid fermentation. AAB are commonly found in various fermented foods such as vinegar, sour beer, fermented cocoa and coffee beans, kefir beverages, kombucha, and sourdough. They interact and cooperate with a variety of microorganisms, resulting in the formation of diverse metabolites and the production of fermented foods with distinct flavors. Understanding the interactions between AAB and other microbes is crucial for effectively controlling and utilizing AAB in fermentation processes. However, these microbial interactions are influenced by factors such as strain type, nutritional conditions, ecological niches, and fermentation duration. In this review, we examine the relationships and research methodologies of microbial interactions and interaction studies between AAB and yeasts, lactic acid bacteria (LAB), and bacilli in different food fermentation processes involving these microorganisms. The objective of this review is to identify key interaction models involving AAB and other microorganisms. The insights gained will provide scientific guidance for the effective utilization of AAB as functional microorganisms in food fermentation processes.

Effect of Mixed Cultures on Microbiological Development in Berliner Weisse Beer

Sour beers play an important role in the brewing market, and their production has been growing exponentially. In light of this, six microorganisms directly related to this class of beer were studied, and the fermentation behavior of six strains used in the past for traditional commercial Berliner Weisse beer production was monitored. The microorganisms used were Lactobacillus brevis, Lactobacillus parabrevis, Brettanomyces bruxellensis, and Brettanomyces anomalus and two strains of Saccharomyces cerevisiae. The six microorganisms were selected in a previous work, and a comparison between single and mixed fermentations was carried out via daily measurements of the fermentation parameters like pH, extract, and cell count during 22 days. The ability to isolate a specific microorganism from a mixed culture was investigated using three commonly used nutrient media and aerobic/anaerobic growth conditions. Both Lactobacillus and Brettanomyces could be isolated; however, the conditions imposed were not sufficient in order to isolate Saccharomyces. Fermentations carried out with LAB and Brettanomyces showed a decrease in Lactobacillus growth if compared to pure fermentations, but no influence on the growth of Brettanomyces could be perceived. In general, fermentations carried out in the presence of Saccharomyces were dominated by this yeast. Its quick growth seems to be responsible for the high end pH values observed as well as the decrease in cell growth for both LAB and Brettanomyces. A decrease in the cell viability of Saccharomyces was followed by an increased growth of the other microorganisms involved, possibly meaning that the molecules released through apoptosis are used by both LAB and Brettanomyces as a valuable nutrient source. The volatile compound concentrations of the first group were higher in fermentations with Saccharomyces, whereas esters’ concentration was higher in fermentations carried out only with Brettanomyces and Lactobacillus. Furthermore, understanding how these microorganisms interact during the fermentation process can help brewers better control production and ensure the consistency in the quality of the final product. The end pH values and acidity reached levels acceptable for Berliner Weisse beer. This innovative approach certainly contributes to the evolution and refinement of the art of brewing.

The Probiotic Paradox: Thriving in High-Hopped Sour Beer

The global craft beer market is witnessing substantial growth, with a particular spotlight on sour beers. These unique brews are created through lactic fermentation initiated by lactic acid bacteria (LAB). However, the presence of hops (Humulus lupulus L.), renowned for their antimicrobial properties, can pose a limitation within the realm of sour beer production. This research aims to explore and assess the potential of sour beer as a probiotic beverage. It involves a comparative analysis of the viability of Lacticaseibacillus paracasei subsp. paracasei F19 (F19) and 431 (L431) in a sour beer with an International Bitterness Unit (IBU) value of 22, alongside examining the expression of genes such as horA, horB, horC, hitA, bsrA, and recA, which are associated with hop resistance. The study encompasses the development of two formulations of sour beers characterized by both high hop content and a robust probiotic count (8.44–8.77 log CFU/mL). The findings suggest that both probiotic strains of F19 and L431, are well-suited to produce sour beers with elevated hop levels, demonstrating excellent stability. Notably, the expression of genes responsible for hop resistance exhibited distinct modulation patterns among the two strains. It appears that a higher concentration of bsrA, as observed in strain L431, may be more effective in mitigating the effects of hop-related stress, potentially surpassing the combined expression of horA and bsrA. Nonetheless, further research is essential to validate this observation in gene expression. F19 and L431 emerge as promising candidates for the development of sour beers with increased hop intensity.

The mechanisms of lactiplantibacillus plantarum J6-6 against iso-α-acid stress and its application in sour beer production

The mechanisms of lactiplantibacillus plantarum J6-6 against iso-α-acid stress and its application in sour beer production

A Review of N-Heterocycles: Mousy Off-Flavor in Sour Beer.

Beer has over 600 flavor compounds and creates a positive tasting experience with acceptable sensory properties, which are essential for the best consumer experience. Spontaneous and mixed-culture fermentation beers, generally classified as sour beers, are gaining popularity compared to typical lager or ale styles, which have dominated in the USA for the last few decades. Unique and acceptable flavor compounds characterize sour beers, but some unfavorable aspects appear in conjunction. One such unfavorable flavor is called "mousy". This description is usually labeled as an unpleasant odor, identifying spoilage of fermented food and beverages. It is related as having the odor of mouse urine, cereal, corn tortilla chips, or freshly baked sour bread. The main compounds responsible for it are N-heterocyclic compounds: 2-acetyltetrahydropyridine, 2-acetyl-1-pyrroline, and 2-ethyltetrahydropyridine. The most common beverages associated with mousy off-flavor are identified in wines, sour beers, other grain-based beverages, and kombucha, which may contain heterofermentative lactic acid bacteria, acetic acid bacteria, and/or yeast/fungus cultures. In particular, the fungal species Brettanomyces bruxellensis are associated with mousy-off flavor occurrence in fermented beverages matrices. However, many factors for N-heterocycle formation are not well-understood. Currently, the research and development of mixed-cultured beer and non/low alcohol beverages (NABLAB) has increased to obtain the highest quality, sensory, functionality, and most notably safety standards, and also to meet consumers' demand for a balanced sourness in these beverages. This paper introduces mousy off-flavor expression in beers and beverages, which occurs in spontaneous or mixed-culture fermentations, with a focus on sour beers due to common inconsistency aspects in fermentation. We discuss and suggest possible pathways of mousy off-flavor development in the beer matrix, which also apply to other fermented beverages, including non/low alcohol drinks, e.g., kombucha and low/nonalcohol beers. Some precautions and modifications may prevent the occurrence of these off-flavor compounds in the beverage matrix: improving raw material quality, adjusting brewing processes, and using specific strains of yeast and bacteria that are less likely to produce the off-flavor. Conceivably, it is clear that spontaneous and mixed culture fermentation is gaining popularity in industrial, craft, and home brewing. The review discusses important elements to identify and understand metabolic pathways, following the prevention of spoilage targeted to off-flavor compounds development in beers and NABLABs.

Transcriptional Analysis of Mixed-Culture Fermentation of Lachancea thermotolerans and Saccharomyces cerevisiae for Natural Fruity Sour Beer

Increasingly high interest in yeast–yeast interactions in mixed-culture fermentation is seen along with beer consumers’ demands driving both market growth and requests for biotechnological solutions that can provide better sensory characteristics. In this study, Lachancea thermotolerans and Saccharomyces cerevisiae with a cell population ratio of 10:1 were inoculated for sour beer fermentation while the process conditions within the brewing industry remained unchanged. With L. thermotolerans producing lactic acid (1.5–1.8 g/L) and bringing down the pH to 3.3–3.4 whilst adding no foreign flavors herein, this study revealed a new natural, fruity sour beer with a soft, sour taste. In this study, the double-yeast mixed-culture fermentation produced more flavor substances than a single-culture process, and plenty of isobutyl acetate and isoamyl acetate enhanced the fruit aroma and balanced the sour beer with a refreshing taste. While playing a positive role in improving the beer’s quality, the double-yeast mixed-culture fermentation developed in this study helps to offer an alternative mass production solution for producing sour beer with the processes better controlled and the fermentation time reduced. The stress responses of the L. thermotolerans during the fermentation were revealed by integrating RNA sequencing (RNA-Seq) and metabolite data. Given that the metabolic flux distribution of the S. cerevisiae during the fermentation differed from that of the non-Saccharomyces yeasts, transcriptional analysis of non-Saccharomyces yeast and S. cerevisiae could be suitable in helping to develop strategies to modulate the transcriptional responses of specific genes that are associated with the aroma compounds released by S. cerevisiae and non-Saccharomyces yeasts. In the case of some non-Saccharomyces yeast species/strains, the diversion of alcoholic fermentation and the formation of a great number of secondary compounds may, in part, account for the low ethanol yield.

Analysis of mousy off-flavor compound 2-Acetyl-tetrahydropyridine using Liquid Chromatography Mass Spectrometry with Electrospray Ionization in sour beer

Mousy off-flavor describes N-heterocycles compounds related to spoilage in the brewing industry. It has also been identified in sour beers through sensory analysis. Therefore, preventing spoilage N-heterocycles development is essential to preserve end-products and obviate economic losses. To this day, no methods or protocols have been reported to identifying mousy off-flavor compounds in a beer matrix. The main objective of this work was to develop a standardized quantification method for 2-acetyl-3,4,5,6-tetrahydropyridine (ATHP) in beer matrix, by Liquid Chromatography Mass Spectrometry with Electrospray Ionization (LC-MS-ESI). Extraction of ATHP in the samples was performed using QuEChERS (quick, easy, cheap, effective, rugged, and safe) technique. Over a dozen different potentially mousy cask-aged sour beers including other spontaneously fermented beverages were provided, based on sensory analysis, to determine the variation in ATHP levels. Results indicated ATHP was found in all the samples, ranging from 1.64 ± 0.06 to 57.96 ± 2.15 µg L−1. Herein, we described our detection method of mousy-off flavor compounds which enables future research to mitigate the occurrence of such defects in fermented beverages matrix.•ATHP content in samples varied from 1.64 ± 0.06 to 57.96 ± 2.15 µg L−1.•The recovery range of ATHP using LC-MS-ESI varied from 71% to 97%.•Basified QuEChERS salting-out procedure is applicable for ATHP extraction from beer and other fermented beverages matrices.

Optimization of beer mixed fermentation using Saccharomyces cerevisiae and Lactobacillus brevis

Mixed fermentation is one of the methods used in sour beer production. The process requires initialization of the fermentation step by well-planned addition of brewing yeast and lactic acid bacteria to slightly hopped wort. The final product’s properties strictly depend on how the microorganisms are pitched and the initial wort composition. The experiment was performed to evaluate the impact of different initial conditions and pitching methods on the mixed fermentation process and the final product’s characteristics. With the aim of limitation of the number of experiments, the Box–Behnken design was applied. Three independent factors were considered while obtaining the response surface: initial extract, bitterness and order of pitching. The final product’s properties: ethanol and lactic acid concentration, appeared to depend strictly on initial conditions and pitching order. Several important observations have been made; for example, it appeared that the presence of LAB does not significantly impact the final ethanol concentration. Optimal conditions for obtaining the maximum or minimum of each quality were calculated using Matlab. Obtained results might improve the sour beer production process while shortening the duration and reducing the usage of ingredients.

Lachancea quebecensis a Novel Isolate for the Production of Craft Beer.

Yeasts are ubiquitously present in different natural sources. Some of these yeasts have interesting characteristics for the production of fermented food products. This study characterized Lachancea thermotolerans and L. quebecensis isolated from insects to determine their brewing potential. The yeasts were evaluated according to their fermentative potential in glucose and maltose-defined media and their resistance to ethanol and hop. Finally, craft beer was elaborated at a laboratory scale (10 L). The yeasts utilized glucose as the only carbon source and produced 3.25 ± 1.77, and 4.25 ± 1.06% (v/v), of ethanol for L. thermotolerans and quebecensis, respectively. While in the maltose-defined medium, ethanol content reached 3.25 ± 0.45, and 3.92 ± 0.36, respectively. The presence of alpha acids and ethanol affected the growth of L. quebecensis, which showed lower growth at 90 IBU and 8 ethanol% (v/v) mixtures. The craft beer brewed with L. quebecensis in monoculture experiments showed fruity flavors associated with ethyl acetate and isoamyl acetate. The ethanol content reached 3.50 ± 0.46% (v/v). The beer pH was 4.06 ± 0.20, with a lactic acid concentration of 1.21 ± 0.05 g/L. The sensory panel identified the beer as "fruity", "floral", "hoppy", "sweet", and "sour". To our knowledge, this is the first time L. quebecensis was reported as a potential candidate for sour beer production with reduced ethanol content.

Sour Beer as Bioreservoir of Novel Craft Ale Yeast Cultures.

The increasing demand for craft beer is driving the search for novel ale yeast cultures from brewing-related wild environments. The focus of bioprospecting for craft cultures is to identify feral yeasts suitable to imprint unique sensorial attributes onto the final product. Here, we integrated phylogenetic, genotypic, genetic, and metabolomic techniques to demonstrate that sour beer during aging in wooden barrels is a source of suitable craft ale yeast candidates. In contrast to the traditional lambic beer maturation phase, during the aging of sour-matured production-style beer, different biotypes of Saccharomyces cerevisiae dominated the cultivable in-house mycobiota, which were followed by Pichia membranifaciens, Brettanomyces bruxellensis, and Brettanomyces anomalus. In addition, three putative S. cerevisiae × Saccharomyces uvarum hybrids were identified. S. cerevisiae feral strains sporulated, produced viable monosporic progenies, and had the STA1 gene downstream as a full-length promoter. During hopped wort fermentation, four S. cerevisiae strains and the S. cerevisiae × S. uvarum hybrid WY213 exceeded non-Saccharomyces strains in fermentative rate and ethanol production except for P. membranifaciens WY122. This strain consumed maltose after a long lag phase, in contrast to the phenotypic profile described for the species. According to the STA1+ genotype, S. cerevisiae partially consumed dextrin. Among the volatile organic compounds (VOCs) produced by S. cerevisiae and the S. cerevisiae × S. uvarum hybrid, phenylethyl alcohol, which has a fruit-like aroma, was the most prevalent. In conclusion, the strains characterized here have relevant brewing properties and are exploitable as indigenous craft beer starters.

Study on the technology of kumquat grapefruit sour beer

A kumquat-grapefruit acid beer was brewed with Australian barley malt, lactobacillu, D17 yeast, Qingdao hop flower, kumquat concentrated juice and grapefruit concentrated juice as raw materials by using special mash and fermentation process. The effects of acidification degree, the addition amount of kumquat and grapefruit juice concentrate and the addition time on beer flavor were studied in this paper. The results showed that when the pH was adjusted to 3.5 by acidification, the sour beer produced by adding 6% kumquat concentrated juice and 4% grapefruit concentrated juice before the CO2 pressure of the tank rised, the beer has white and exquisite foam with prominent fruit aroma, especially those of kumquat and grapefruit. The beer body was clear and transparent, with about 5% residual sugar. The sweet and sour taste of the beer was balanced. The beer contained unique nutrients of kumquat and grapefruit also, and was especially suitable to drink for brewers and ladies.

Sequentially Pitching Lactic Acid Bacteria and Active Dry Yeasts for Sour Beer Production

Lactic acid bacteria are key constituents in the souring process of traditional acidic beers, resulting from spontaneous or mixed fermentations. The development of such beer styles provides a balanced and complex-flavored beverage but is time-consuming and not easily repeatable. New time-efficient alternatives include successively pitching a souring bacteria and yeast, such that both have optimal fermentation conditions. In this study, each microorganism is assessed to comprehend its individual impact on the fermentation process (acidification and fermentation rates, attenuation, ethanol production) and its influence on physicochemical and organoleptic parameters. Lactic acid bacteria are responsible for the wort’s acidification, reaching a pH of 3.5 within 48 h, by converting up to 5 g/L of simple sugars into organic acids and other by-products. The two-step acidification and fermentation process alters the color and bitterness, the latter decreasing from an initial iso-α-acid concentration of 29 mg/L to an average of 13 mg/L. The sequential method shows that the decarboxylation and subsequent reduction of hydroxycinnamic acid precursors into their vinyl and ethyl derivatives is strain dependent. All tested lactic acid bacteria appear to possess the decarboxylation enzymes. However, only Lactiplantibacillus plantarum can reduce vinylphenols into ethylphenols.

Study on preparation technology for mango sour beer [pdf

Study on preparation technology for mango sour beer [pdf

Study on preparation technology for mango sour beer

Background. Mango sour beer was made from mango and malt. A sensory score was used as an index to study the effects of main fermentation temperature, Lactobacillus plantarum addition amount and fermentation tank pressure on the taste of mango sour beer. Material and methods. Based on the single-factor experiment, a response surface methodology was used to optimize the preparation process of mango sour beer. Results. The results showed that the optimum fermentation conditions were as follows: the main fermentation temperature was 25℃, the Lactobacillus plantarum content was 0.25 kg/KL and the fermenting pressure was 0.15 MPa. Conclusion. At this point, the mango sour beer was bright and golden, with a clear and transparent body, a rich fruit flavor, an outstanding sour flavor and a mellow wheat flavor, which provides a reference point for the subsequent optimization of the production process and the production of mango sour beer.

The combination of omics strategies to evaluate starter and probiotic strains in the Catharina sour Brazilian-style beer

Catharina sour, the first internationally recognized Brazilian beer, is characterized by fermentation with lactic acid bacteria (LAB), which may have probiotic potential, and the addition of fruit juice. This study aimed to evaluate the use of the starter Streptococcus thermophilus TH‐4 (TH‐4) and the probiotics Lacticaseibacillus paracasei F19 and 431, associated with Saccharomyces cerevisiae US-05, in the absence (control)/presence of passion fruit or peach juices. Evaluation proceeded during fermentation and storage by enumeration using pour-plate and qPCR; gene expressions of hop resistance; proteome by Liquid Chromatography Tandem Mass Spectrometry (LC-MS/MS); and odor, flavor, and metabolome by Headspace Solid-Phase Microextraction (HS-SPME), coupled with the gas chromatography–mass spectrometry (GC–MS) analysis. We concluded that the strains studied are recommended for applications in sour beers, due to the presence of defense mechanisms like membrane adhesion and H + pump. Furthermore, HS-SPME/GC-MS indicated that the strains may contribute to the beer flavor and odor.

Lachancea thermotolerans, an Innovative Alternative for Sour Beer Production

The interest in and growth of craft beer has led to an intense search for new beers and styles. The revival of traditional styles has sometimes been hampered by the use of microorganisms such as lactic acid bacteria. Therefore, studies on alternative yeasts for the production of this style of beer have increased. In this work and together with previous studies carried out with yeasts isolated from Madrid agriculture (from grapes, must, wine, vineyards and wineries), the capacity of 10 yeast strains, belonging to the genus Lachancea thermotolerans, for the production of sour beer has been determined. For this purpose, different fermentation scale-ups (100 mL, 1 L and 100 L) have been performed and their fermentation capacity, aroma compound production (33 volatile compounds by GC), organoleptic profile (trained tasting panel and consumers), melatonin production (HPLC) and antioxidant capacity have been studied. Beer fermented with yeast strain CLI 1232 showed a balanced acidity with a fruity aromatic profile and honey notes. On the other hand, the beer fermented with strain 1-8B also showed a balanced acidity, but less fruity and citric flavour than CLI 1232 strain. Finally, the yeast strain selected by the consumers (CLI 1232) was used for beer production at industrial scale and the market launch of a sour beer.