Influence of pollen addition on mead elaboration: Physicochemical and sensory characteristics

Mead (honey-wine) results from the alcoholic fermentation of diluted honey using a wine yeast strain. However, mead elaboration can be hampered by several problems, including delayed or arrested fermentation, production of an unpleasant aroma, poor quality and inconsistency of the final product. These difficulties are due to honey’s low nutrient content, its natural antifungal components, and the inability of the yeast strain to adapt to these unfavourable growth conditions. In this study, we evaluated the results of adding pollen at concentrations ranging from 10 to 50 g/l as a fermentation activator to improve the fermentation kinetic and the quality of meads. The effect of pollen addition on the honey must, fermentation kinetics, physicochemical characteristics, aroma profiles and sensorial aspects of the obtained meads were evaluated. The results showed that pollen addition improved fermentation rates, alcohol yields, and the final characteristics of meads. An increase in the volatile contents of the meads and an improved sensory profile was observed with pollen addition; however, this improvement was not correlated with the concentration of pollen. The adequate dose of pollen was determined by the final characteristics and sensory profile of the meads.     

Mead production: effect of nitrogen supplementation on growth,fermentation profile and aroma formation by yeasts in mead fermentation

Mead is an alcoholic beverage, produced since ancient times, resulting from an alcoholic fermentation of diluted honey by yeasts. When it is produced in a traditional manner, mead producers can encounter several problems related to a lack of essential nutrients, such as available nitrogen. Thus, the aim of this study was to evaluate the effect of nitrogen addition to honey‐must on the fermentation performance of two Saccharomyces cerevisiae wine yeasts, QA23 and ICV D47, as well as on the mead composition and production of volatile aroma compounds. A portion of honey‐must was supplemented with diammonium phosphate (DAP) to achieve the nitrogen concentration required by yeast to complete alcoholic fermentation. The supplementation with DAP reduced the fermentation length to around 7 days, but not all sugars were fully consumed, suggesting that other factors could be interfering with yeast growth. For both yeasts the specific growth rate and final biomass were higher in musts supplemented with DAP. Mead final composition was similar under the two experimental conditions. Analysis of the volatile profile revealed that the concentrations of the volatile fatty acids and volatile phenols were higher in meads supplemented with DAP. The concentrations of ethyl hexanoate, ethyl octanoate and isoamyl acetate were above their perception threshold and were higher in meads supplemented with DAP, which could contribute to the enhancement of the fruity character. This study could be useful for the optimization of mead production and quality improvement. Copyright © 2015 The Institute of Brewing & Distilling     

High-cell-density fermentation of Saccharomyces cerevisiae for the optimisation of mead production

Mead is a traditional drink that contains 8%–18% (v/v) of ethanol, resulting from the alcoholic fermentation of diluted honey by yeasts. Mead fermentation is a time-consuming process and the quality of the final product is highly variable. Therefore, the present investigation had two main objectives: first, to determine the adequate inoculum size of two commercial wine-making strains of Saccharomyces cerevisiae for the optimisation of mead fermentation; and second, to determine if an increase in yeast pitching rates in batch fermentations altered the resulting aroma profiles. Minor differences were detected in the growth kinetics between the two strains at the lowest pitching rate. With increasing pitching rates net growth of the strain ICV D47 progressively decreased, whereas for the QA23 the increasing inoculum size had no influence on its net growth. The time required to reach the same stage of fermentation ranged from 24 to 96 h depending on the inoculum size. The final aroma composition was dependent on the yeast strain and inoculum size. Fourteen of the twenty-seven volatile compounds quantified could contribute to mead aroma and flavour because their concentrations rose above their respective thresholds. The formation of these compounds was particularly pronounced at low pitching rates, except in mead fermented by strain ICV D47, at 10⁶ CFUs/mL. The esters isoamyl acetate, ethyl octanoate and ethyl hexanoate were the major powerful odourants found in the meads. The results obtained in this study demonstrate that yeast strain and inoculum size can favourably impact mead's flavour and aroma profiles.     

Effect of Saccharomyces cerevisiae cells immobilisation on mead production

Mead is a traditional alcoholic beverage obtained by the fermentation of diluted honey performed by yeasts. In this work the potential of application of immobilised yeast cells on single-layer Ca-alginate or double-layer alginate–chitosan for mead production was assessed for the first time. The meads produced either with entrapped or free cells were evaluated in terms of quality and aroma profile. The immobilisation procedure had no adverse effect on cell viability, since minor differences were found in fermentation kinetics among the strains and immobilisation systems. The double-layer alginate-chitosan had no advantage compared with the single-layer Ca-alginate, as the number of free cells in the medium, resulting from cell leakage, was similar. Although meads obtained with entrapped yeast cells presented less ethanol and glycerol and more acetic acid, it exhibited larger amounts of volatile compounds. Immobilised cells produced meads with more compounds with fruity characteristics, such as ethyl octanoate and ethyl hexanoate; however the concentrations of undesirable compounds in such meads were also higher. The effect of immobilisation on the aroma profile was important, but the strain contribution was also of major importance. Thus, the sensory analysis of final product gives an important insight on the overall quality.     

Optimization of honey-must preparation and alcoholic fermentation by Saccharomyces cerevisiae for mead production

Mead fermentation is a time-consuming process, often taking several months to complete. Despite of the use of starter cultures several problems still persist such as lack of uniformity of the final products, slow or premature fermentation arrest and the production of off-flavors by yeast. Thus the aim of this study was to optimize mead production through the use of an appropriate honey-must formulation to improve yeast performance alcoholic fermentation and thereby obtain a high quality product. Honey-must was centrifuged to reduce insoluble solids, pasteurized at 65 °C for 10 min, and then subjected to different conditions: nitrogen supplementation and addition of organic acids. Although the addition of diammonium phosphate (DAP) reduced fermentation length, it did not guarantee the completeness of the fermentation process, suggesting that other factors could account for the reduced yeast activity in honey-must fermentations. Sixteen yeast-derived aroma compounds which contribute to the sensorial quality of mead were identified and quantified. Global analysis of aromatic profiles revealed that the total concentration of aroma compounds in meads was higher in those fermentations where DAP was added. A positive correlation between nitrogen availability and the levels of ethyl and acetate esters, associated to the fruity character of fermented beverages, was observed whereas the presence of potassium tartrate and malic acid decreased, in general, their concentration.This study provides very useful information that can be used for improving mead quality.     

Effect of Heat Treatment on Antioxidant Capacity and Flavor Volatiles of Mead

ABSTRACT: The objective of this study was to evaluate the effects of heat processing on the antioxidant capacity of mead (honey wine). Soy and buckwheat honey musts were subjected to 2 heat treatments and fermented into wine. Total phenolic concentration was determined. High-performance liquid chromatography (HPLC) phenolic profiling was performed on the methanol fraction of Amberlite extraction. Antioxidant capacity was evaluated using the oxygen radical absorbance capacity (ORAC) assay. Changes in volatile components were evaluated by headspace-solid phase microextraction/gas chromatography-mass spectrometry (H-SPME/GC-MS). ORAC values of experimental meads (3.62 m M Trolox equivalent) were comparable to those of commercial white wine (3.66 m M Trolox equivalent). No significant difference in antioxidant capacity due to heat treatment or honey type was observed. There was no difference in total phenolics between heat treatments in buckwheat mead; however, soy mead made from high-heated must had significantly greater phenolic concentration than the gently heated mead (α= 0.05). Linear regression analysis indicated a strong positive correlation between total phenolic concentration and antioxidant capacity by ORAC ( r = 0.9077; P < 0.0001). HPLC analysis of phenolic profiles in the methanol fractions of Amberlite extraction of the meads indicated significantly higher levels of certain phenolics as a result of the high-heat process in buckwheat mead, but not in soy mead. Differences in volatile components that potentially impact flavor were noted between high and low heat treatments. Results of this study suggest dramatic heat treatments that are often avoided because their flavor impact in mead production have the potential to alter the antioxidant capacity of mead by changing phenolic profiles.     

Determination of some individual phenolic compounds and antioxidant capacity of mead produced from different types of honey

The aim of this study was to promote mead (honey wine) production by testing the appropriateness of different honey varieties which were obtained from the flora of Turkey for the production of mead. Cotton‐Mezda (Gossypium spp.), pine (Pinus spp.) and flower honeys were used in the fermentation studies. Pure culture Saccharomyces cerevisiae yeasts were used in the mead studies. While determining the phenolic contents of meads via HPLC method, five phenolic acids and two flavonoids were used as standards with known antioxidant properties. In addition, total antioxidant activity was analysed in meads using trolox equivalent antioxidant capacity (TEAC). Among the phenolic acids, the most abundant one was protocatechuic acid (75.12–179.03 μg/L) and among flavonoids it was catechin (10.38–125.55 μg/L). It was found out that total phenolic content ranged from 103.56 GAE to 167.89 GAE mg/L. Copyright © 2017 The Institute of Brewing & Distilling     

The influence of yeast immobilization on selected parameters of young meads

Mead is a traditional product derived from the fermentation of diluted honey. The main purpose of the study was to determine the influence of Saccharomyces cerevisiae immobilization in calcium alginate, κ‐carrageenan and lightly crosslinked poly(acrylic acid) on fundamental enological parameters and concentrations of formic and acetic acids (SPME‐GC) in fermented mead worts. Ionic hydrogels, especially poly(acrylic acid) and κ‐carrageenan, increase the ethanol concentration (up to 18% v /v) in young meads and reduce titratable (35%) and volatile acidity (67%), primarily by decreasing acetic acid synthesis with the use of immobilized cells of yeast. The surface immobilization on poly(acrylic acid) resulted in a reduction of the time of fermentation by about 25 days. The immobilization of yeast inside calcium alginate and κ‐carrageenan gels extended the time of preliminary fermentation and reduced the degree of fermentation in samples with calcium alginate. Copyright © 2017 The Institute of Brewing & Distilling     

Mead production: fermentative performance of yeasts entrapped in different concentrations of alginate

Mead is an alcoholic drink known since ancient times, produced by yeast fermenting diluted honey. However, the production of mead has suffered in recent years, partially owing to the lack of scientific progress in this field. In this study, two strains of Saccharomyces cerevisiae, QA23 and ICVD47, were immobilized in 2 or 4% (w/v) alginate beads to assess the most effective alginate concentration for yeast immobilization to produce mead. Neither of the alginate concentrations was able to prevent cell leakage from the beads. The fermentation length was 120 h for both yeast strains. In all cases, at the end of the fermentation, the number of cells entrapped in the beads was higher than the number of free cells, and the total 4% alginate bead wet weight was significantly higher than the 2% alginate bead wet weight. In addition, the evaluation of mead quality showed that the yeast strain had significantly more influence on the physicochemical characteristics than the alginate concentration. Although the yeasts immobilized in the two alginate concentrations were able to perform the fermentation, further research is needed in order to understand the evolution of the yeast population inside the beads throughout the fermentative process. Copyright © 2014 The Institute of Brewing & Distilling     

Physicochemical property changes during the fermentation of longan (Dimocarpus longan) mead and its aroma composition using multiple yeast inoculations

Mead is a traditional drink obtained from diluted honey with the appropriate yeast inoculations. However, the production of mead is time‐consuming, and the process often takes several months to accomplish. During the elaboration of mead, sluggish and stuck fermentations often occur and lead to the production of undesirable flavours. To improve the condition of longan mead production in Taiwan, multiple yeast inoculations with different strains of yeast were examined in this study. The physicochemical properties and mead aroma compounds were measured. Longan mead without multiple inoculations, but with added yeast nutrients, displayed a sharp increase in ethanol content compared with the inoculated groups without yeast nutrients. However, the physicochemical properties of every mead reached almost the same endpoint at the end of the fermentation time (day 22). Moreover, differences among the groups were observed in their aroma compounds. Some aroma compounds, such as 2‐methyltetrahydrothiophen‐3‐one, 1‐propanol and 2‐methylbutyl acetate, were only found in the double‐inoculation groups, whereas other aroma compounds, such as phenethyl acetate and linalool, were only found in the single‐yeast inoculation groups. These results demonstrate that multiple inoculations have a positive effect on longan mead fermentation. This technique could assist local producers in elaborating a longan mead of acceptable quality. Copyright © 2013 The Institute of Brewing & Distilling     

蜂蜜酒酿造工艺技术的研究与应用

蜂蜜酒是以蜂蜜为原料,经过发酵后所获得的酒精饮料。为了酿造较高酒精度的优质蜂蜜酒,从蜜源挑选、蜜汁灭菌、溶氧操作、选育优良酵母菌株。确定蜂蜜酒发酵过程中流加的次数,流加的量和流加间隔时间,并建立流加的方程。适温发酵、过滤除浊、精心调配等工序,取得蜂蜜酒酿造最佳技术参数。     

Production and characterization of mead from the honey of Melipona scutellaris stingless bees

Mead is a traditional alcoholic beverage obtained by fermenting must and can offer a solution to honey over‐production and a way of valorizing honey of lower quality. The purpose of this study was to produce and characterize mead with different levels of sugars and alcohol obtained from honey from Melipona scutellaris. The honey used for mead preparation was analysed in order to ensure that it met the required quality standards. It was found that the alcoholic content and volatile acidity were outwith the limits established by Brazilian law. Mead legislation is based on the product obtained from Apis mellifera (‘honey bee’) honey and these results indicate the need to re‐evaluate the standards established for this product in order to incorporate mead produced from honey from stingless bees of the genus Melipona. Copyright © 2018 The Institute of Brewing & Distilling     

蜂蜜中酵母菌的分离鉴定及蜂蜜酒的研制

采用稀释涂布平板法分离纯化蜂蜜样品中的菌株,并对其进行鉴定,并对蜂蜜酒进行了研制并对其挥发性成分进行测定。结果表明,从12份采自云南文山的蜂蜜样品中分离出5株酵母菌（Y3、Y5、Y7、Y9、Y10）,经5.8S rDNA-ITS 序列分析,结合形态学特征初步鉴定为暹罗接合酵母（Zygosaccharomyces siamensis）。5株菌用于蜂蜜稀释液发酵,其中菌株Y3发酵的蜂蜜酒酒香和果香浓郁,是较好的蜂蜜酒发酵菌株。以同时蒸馏萃取（SDE）-气质联用（GC-MS）技术对菌株Y3发酵的蜂蜜酒风味成分进行分析,共检测出41种挥发性成分,主要成分为2-苯乙醇（58.11%）和异戊醇（25.43%）。     

Production and Antioxidative Activity of Mead Made From Honey and Black Rice (Oryza sativa var. Indica cv. Shiun)

In order to brew a red‐coloured alcoholic beverage having antioxidative activity, mead was made from honey and black rice grains. Alcoholic beverages made from honey and black rice grains contained around 10% (v/v) ethyl alcohol and a higher amount of phenolic compounds. Alcoholic beverages made from Chinese milk vetch honey and black rice grain contained anthocyanin and were red in colour similar to pink wine. The amount of total phenolic compounds present in the beverages made using Chinese milk vetch honey and buckwheat honey was 0.2 and 0.4 mg/mL, respectively. The DPPH radical scavenging activity of the mead made from the black rice grains was higher than that of beverages made from polished rice. The inhibitory activity of lipid peroxidation of the mead made from black rice grains was also higher.     

Isolation and Characterisation of Sri Lankan Yeast Germplasm and Its Evaluation for Alcohol Production

Use of inferior yeast cultures represents one of the reasons for low fermentation efficiencies in Sri Lankan alcohol distilleries that use sugarcane molasses. The present study isolated and characterised yeast strains found in natural environments in Sri Lanka and evaluated their performance under laboratory conditions in an effort to select superior strains for industrial fermentations. Yeasts were characterised based on morphological and physiological features such as sugar fermentation and nitrate assimilation. Ethanol production, alcohol tolerance and growth rate of the most promising strains were monitored following laboratory fermentations of molasses. Over a thousand yeast cultures were collected and screened for fermentative activity and a total of 83 yeast isolates were characterised as higher ethanol producers. Most of these belonged to the genus Saccharomyces. Certain strains produced over 10% (v/v) alcohol in molasses media during 72 h laboratory fermentations. Only two strains, SL‐SRI‐C‐102 and 111, showed an appreciable fermentation efficiency of about 90%. The latter strain produced the highest level of ethanol, 11% (v/v) within a 48 h fermentation and exhibited improved alcohol tolerance when compared with the baker's yeast strains currently used in Sri Lankan alcohol distilleries. This study highlights the benefits of exploiting indigenous yeasts for industrial fermentation processes.     

Replacement of sulfur dioxide by lysozyme and oenological tannins during fermentation: influence on volatile composition of white wines

BACKGROUND: In recent years the use of sulfur dioxide, a commonly used additive in winemaking, has been questioned because of its toxic effects on human health. Studies have been conducted to find alternatives that can effectively substitute for this additive in all its various technological functions. In previous work, lysozyme and oenological tannins were found as possible substitutes in controlling bacterial undesirable fermentations and phenolic oxidation. However, data on the volatile composition of wines obtained by that protocol are lacking. In this work, the effects on volatile composition of white wines by the substitution of SO₂ during fermentation with lysozyme and tannin were studied. At the same time, the technological performance of two strains of yeast that produce low amounts of SO₂ were evaluated. RESULTS: The results showed that both SO₂ and lysozyme prevented the development of undesirable bacterial fermentations. The study of volatile compounds shows differences in the alcohol, acid and ester contents among the final products: wines fermented with strain 1042 and lysozyme had higher total alcohol concentration, while the addition of SO₂ promoted higher production of 3‐methyl‐1‐butanol, 3‐methylthio‐1‐propanol, phenylethyl alcohol and 4‐hydroxy‐benzenethanol. Esters, as a total, were influenced by the different strain and tannins addition, while amounts of medium‐chain fatty acid ethyl esters and their corresponding fatty acids were found in higher amounts in wines coming from fermentations with lysozyme. The sensory analysis revealed a preference for wines to which lysozyme and tannins had been added. CONCLUSION: The data suggest that the addition of lysozyme and oenological tannins during alcoholic fermentation could represent a promising alternative to the use of SO₂ and for the production of wines with reduced content of SO₂. The composition of the volatiles in the final wines was affected by the different vinification protocols (mainly with regards to alcohols and ethyl esters). Copyright © 2009 Society of Chemical Industry     

The temperature dependent functionality of Brettanomyces bruxellensis strains in wort fermentations

In recent years, Brettanomyces bruxellensis has found increasing application in brewery fermentations. Indeed, B. bruxellensis contributes to the flavour profile of many Belgian beers, typically during secondary or spontaneous fermentation. In North America, the yeast is used in primary fermentation to produce beers with ‘Brett’ characteristics with ‘fruity’ and/or ‘funky’ sensory profiles associated with the production of volatile esters and phenols. However, little is understood about the factors that influence flavour metabolite production or fermentation rate in this yeast. Here, the impact of temperature is reported on fermentation efficiency, flavour metabolite production and carbon utilisation of one commonly used and eight poorly characterised B. bruxellensis strains during wort fermentation. A high degree of strain and temperature‐dependent variability was found in fermentation efficiency and metabolite production amongst B. bruxellensis strains. Further, fermentation efficiency and carbon utilisation were temperature dependent, while ester production increased at higher temperature and phenol production was strain and temperature independent. These results indicate significant strain and temperature dependent variation, suggesting the potential application of strain variability as a tool to achieve product diversity in B. bruxellensis primary fermentations. © 2019 The Institute of Brewing & Distilling     

Application of fluorescence in situ hybridisation (FISH) to the analysis of yeast population dynamics in winery and laboratory grape must fermentations

To analyse the yeast population diversity during wine fermentations, specific fluorescein-labelled oligonucleotide probes targeted to the D1/D2 region of the 26S rRNA of different yeast species known to occur frequently in this environment were designed and tested with reference strains. The probes were then used to identify wine must isolates and to follow, in combination with plate counts, the evolution of yeast populations in two winery fermentations of white and red grape musts. In both cases, a high diversity of non-Saccharomyces yeast species was detected, including Candida stellata, Hanseniaspora uvarum, H. guilliermondii, Kluyveromyces marxianus, K. thermotolerans and Torulaspora delbrueckii. Some of these species (e.g., K. marxianus, K. thermotolerans and T. delbrueckii) were present in significant amounts during the tumultuous fermentation stage, despite the predominance of Saccharomyces cerevisiae cells following the inoculation of the wine musts with a starter strain. To further clarify the yeast population dynamics at the late phase of the fermentations, and because winery conditions do not allow a reliable control of experimental variables, strains isolated from the industrial musts were used to conduct two laboratory microvinifications in synthetic grape juice, using different ratios of S. cerevisiae/non-Saccharomyces in the inocula. Under these conditions, the results were similar to those obtained in the winery, showing a yeast profile with mixed species throughout the first fermentation stage, i.e. until about 40-50% of the total sugar was consumed. Non-Saccharomyces yeasts were outgrown by S. cerevisiae only after ethanol reached concentrations around 4-5% (v/v), which argues in favour of a potential important role of non-Saccharomyces in the final organoleptic characteristics of the wine.     

Influence of SO<Subscript>2</Subscript> on the evolution of volatile compounds through alcoholic fermentation of must stabilized by pulsed electric fields

The aim of this work was to study the influence of sulphur dioxide (SO₂) on the formation of volatile compounds by yeast through wine alcoholic fermentation. Thus Parellada must was microbiologically stabilized using a pulsed electric field (PEF) treatment and inoculated with Saccharomyces cerevisiae Na33 strain. Fermentation was carried out with or without SO₂ and the results showed that the evolution of the volatile compounds profile throughout the process was similar. The content of volatile acids in wine obtained by using sulphur dioxide was not significantly different from that fermented without adding the compound. However, the final content of total alcohols and esters was significantly different even thought the differences were small. Consequently, when grape must is treated by PEF the sulphur dioxide concentration usually used in winemaking could be reduced to safer levels or even eliminated without an important effect on the volatile compounds content of the final product. Therefore, the absence of sulphur dioxide should not have a negative impact on the sensory characteristics of wine.     

Influence of addition of ammonium and different amino acid concentrations on nitrogen metabolism in spontaneous must fermentation

The effect of the addition of different amino acid concentrations in must on yeast nitrogen metabolism during alcoholic fermentation was studied. To do this, fermentations of Mazuelo must, poor in nitrogen compounds, were carried out. Ammonium and different concentrations of amino acids (0, 45, 120, 250 and 450 mg/l) were added to the must. Addition of 45, 120 and 250 mg/l of proteic amino acids to the must increased the rate of fermentation. Proline was mainly consumed in fermentations with smaller amounts of amino nitrogen and, at the same time, this amino acid showed the highest residual concentration in the final wines. The consumption of other proteic amino acids was directly proportional to their concentration in the musts, with the exception of leucine and isoleucine that were synthesized. However, a difference in the percentages of the amino acids consumed by the yeasts was observed. The percentages of aspartic acid, alanine and arginine consumed were higher in the fermentations supplemented with amino acids than in the fermentation where only ammonium was added. The percentages of tyrosine and phenylalanine consumed gradually increased with increase of their initial concentration.