Impact of two Williopsis yeast strains on the volatile composition of mango wine

The aim of the research was to study the volatile composition of mango wine fermented with two Williopsis yeast strains: Williopsis saturnus var. mrakii NCYC500 and W. staurnus var. suaveolens NCYC2586. Thirty terpenoids, twenty‐six esters, ten alcohols, nine acids, seven aldehydes and ketones, two ethers, two phenols and one sulphur compound were identified in the mango wine fermented with strain NCYC500, while twenty‐seven terpenoids, thirty esters, eleven alcohols, eight acids, eight aldehydes and ketones, three ethers, two phenols, one sulphur compound and one furan were detected in the mango wine fermented with the strain NCYC2586. The kinetic changes, final concentrations and odour activity values of major volatiles were compared between the two Williopsis yeast strains and also with other yeast reported in the literature. The results showed that Williopsis yeast strains NCYC500 and NCYC2586 were high producers of acetic acid and acetate esters, but low producers of medium‐ to long‐chain fatty acids and their corresponding ethyl esters. Unlike mango wine fermented with Saccharomyces cerevisiae, most terpenoids derived from mango juice were retained in the resultant mango wine fermented with the two Williopsis yeast strains, suggesting the mango wine could retain the aromatic hints of fresh mango.     

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     

Evolution of volatile compounds in papaya wine fermented with three Williopsis saturnus yeasts

The changes in volatile compounds during papaya juice fermentation with three Williopsis saturnus yeasts were investigated in this study. Time‐course papaya juice fermentations were carried out using three Williopsis saturnus yeasts: W. saturnus var. mrakii NCYC2251, W. saturnus var. saturnus NCYC22 and W. saturnus var. sargentensis NCYC2727. Changes in yeast cell population, Brix and pH were similar among the three yeasts, which preferentially utilised glucose over fructose while partially degrading l ‐malic acid. A range of volatile compounds were produced during fermentation including fatty acids, alcohols and esters with esters being the most abundant volatile compounds produced. Benzyl isothiocyanate, butyric acid, 2‐ethylhexanol, benzaldehyde and β‐damascenone present in the papaya juice were metabolised to trace levels during fermentation. There were significant variations among the three yeasts in their ability to produce and metabolise volatile compounds during fermentation. The study suggests that papaya juice fermentation with W. saturnus yeasts is able to result in the formation of a more complex aroma compounds.     

Construction of a brewing yeast expressing the glucoamylase gene STA1 by mating

Standard brewing yeast cannot utilize larger oligomers or dextrins, which represent about 25% of wort sugars. A brewing yeast strain that could ferment these additional sugars to ethanol would be useful for producing low‐carbohydrate diabetic or low‐calorie beers. In this study, a brewing yeast strain that secretes glucoamylase was constructed by mating. The resulting Saccharomyces cerevisiae 278/113371 yeast was MAT a/α diploid, but expressed the glucoamylase gene STA1 . At the early phase of the fermentation test in malt extract medium, the fermentation rate of the diploid STA1 strain was slower than those of both the parent strain S. cerevisiae MAFF113371 and the reference strain bottom‐fermenting yeast Weihenstephan 34/70. At the later phase of the fermentation test, however, the fermentation rate of the STA1 yeast strain was faster than those of the other strains. The concentration of ethanol in the culture supernatant of the STA1 yeast strain after the fermentation test was higher than those of the others. The concentration of all maltooligosaccharides in the culture supernatant of the STA1 yeast strain after the fermentation test was lower than those of the parent and reference strains, whereas the concentrations of flavour compounds in the culture supernatant were higher. These effects are due to the glucoamylase secreted by the constructed STA1 yeast strain. In summary, a glucoamylase‐secreting diploid yeast has been constructed by mating that will be useful for producing novel types of beer owing to its different fermentation pattern and concentrations of ethanol and flavour compounds. Copyright © 2017 The Institute of Brewing & Distilling     

Identification and characterization of yeast isolated from the elaboration of seasoned green table olives

The purpose of this study was to investigate the yeast population during the processing of green table olives. In the fresh olives, yeast were found at concentrations of around 3.0 log cfu/g, with Cryptococcus spp. being predominant. In the brine, the yeast concentrations were greater than 4.9 log cfu/ml, with Pichia anomala, Kluyveromyces marxianus, and Saccharomyces cerevisiae being the predominant species. Unlike the yeast isolated from the fresh olives, the strains obtained from the olive brine mostly showed low pectolytic but high catalase activities. Some of these strains also exhibited other biochemical desirable properties for the fermentation of green table olives, including their lipolytic activities and their assimilation or production of organic acids in the brine. Seven strains in particular of P. anomala, K. marxianus, S. cerevisiae, and Candida maris showed the best properties for use in trials as starter culture in pilot fermenters.     

添加淋饭酒母对液态法酿制清爽型山兰黄酒品质及风味的影响

采用淋饭酒母取代传统的纯种酒母酿造清爽型山兰黄酒。结果表明,液态法酿制过程中,与纯种酒母发酵相比,添加3％淋饭酒母发酵平缓,所得山兰黄酒香气浓郁。通过电子鼻主成分及判别因子分析,能很好的区分纯种酒母发酵与淋饭酒母发酵的酒样,纯种酒母发酵的酒样香气与3％淋饭酒母发酵的酒样香气差异显著。通过氨基酸成分分析,淋饭酒母发酵的山兰黄酒游离氨基酸总量更低,口感更加清爽。     

Random and targeted gene integrations through the control of non‐homologous end joining in the yeast Kluyveromyces marxianus

Kluyveromyces marxianus DMKU3‐1042 is a thermotolerant yeast strain suitable for high‐temperature ethanol fermentation and genetic engineering with linear DNA. We have developed a highly efficient random gene integration method with a frequency that exceeds 2.5 × 10⁶ transformants/µg linear DNA, a figure comparable to what is observed with autonomously replicating plasmid transformation in Saccharomyces cerevisiae. To establish the mechanism of random integration in DMKU3‐1042, we identified and deleted the K. marxianus KU70 gene, which is known to be involved in the non‐homologous end‐joining (NHEJ) pathway. In yeast lacking KU70, high‐frequency non‐homologous gene integration was abolished and the Kmku70 mutants showed 82–95% homologous gene targeting efficiencies using homologous sequences of 40–1000 bp. These results indicate that the highly efficient NHEJ pathway can be utilized with random gene disruption techniques such as transposon mutagenesis and plasmid‐free gene manipulations in K. marxianus. Copyright © 2009 John Wiley & Sons, Ltd.     

Intracellular metabolite profiling of industrial yeast and the synthesis of flavour compounds in beer

Beer quality is largely guaranteed by a rational ratio of different flavour compounds. In this study, we present intracellular metabolic profiles in the light of lager beer flavour compound synthesis on an industrial scale. A total of 62 intracellular metabolites were identified and quantitatively analysed. Among these, amino acid was identified as the most dominant category of metabolites. We also report gene expression profiling of relevant genes encoding six representing amino acids, including serine, alanine, lysine, valine, leucine and asparagine. Nine genes involved in the amino acid synthesis – ALT1 , ALT2 , ASN1 , ASN2 , BAT1 , SER2 , SHM1 , SHM2 and LYA1 – displayed significantly higher level of expression at various fermentation stages in an industrial lager fermentation. However, concentrations of the recovered amino acids decreased towards the end of fermentation. Our results suggested that the defined amino acids could be assimilated by the yeast to be utilized for synthesis of aroma‐active metabolites at different stages in fermentation. Copyright © 2017 The Institute of Brewing & Distilling     

Simultaneous utilization of ammonia,free amino acids and peptides during fermentative growth of Saccharomyces cerevisiae

The efficiency of nitrogen use by yeast is one of the key determinants of the successful completion of alcoholic fermentations. In this work the growth of Saccharomyces cerevisiae S288c in a synthetic medium containing ammonia and free amino acids, supplemented with yeast hydrolysate, was studied. Experiments with ¹⁵NH₄Cl and ¹⁵N‐labelled yeast hydrolysate were carried out to gain insight into which of these three classes of assimilable nitrogen sources yeast cells prefer. Co‐consumption of all three sources was observed; approximately 40% of the total nitrogen in the yeast protein fraction originated from yeast hydrolysate, while free amino acids and ammonia contributed 40 and 20%, respectively. The results indicate that several amino acids are more readily obtained from peptides, most likely when the uptake of their free forms is competitively inhibited and/or repressed. During the second half of each fermentation, a decrease in the incorporation of yeast hydrolysate‐derived nitrogen was observed. These results highlight the nutritional role of peptides in various yeast fermentations. Copyright © 2016 The Institute of Brewing & Distilling     

Fermentative capacity of Kluyveromyces marxianus and Saccharomyces cerevisiae after oxidative stress

Volatile compound production during alcoholic fermentation has been studied in the production of many beverages. Temperature, yeast strain, nutrients and pH have been identified as important factors in the production of volatile compounds. In addition, other factors could influence this production during the fermentation process as well. Oxidative stress could occur during yeast biomass production because oxygen is an essential nutrient that is added to the growth medium. The fermentation parameters and the volatile compound production of one Saccharomyces cerevisiae strain (MC4) and two Kluyveromyces marxianus strains (OFF1 and SLP1) were evaluated in relation to fermentation parameters after oxidative stress induced by hydrogen peroxide or menadione. These yeasts were compared with S. cerevisiae W303–1A and showed significant differences in ethanol production, ethanol yield and maximum ethanol production rate. K. marxianus (OFF1) showed better fermentative capacity after oxidative stress. The higher alcohol production decreased after oxidative stress by >35% after 72 h fermentation time, and the amyl alcohol decreased at a higher level (>60%); however, the isobutanol production increased after oxidative stress between 1.5 and 4 times. The yeasts produced significant concentrations of esters however ethyl lactate, ethyl caprylate and the ethyl caproate were not detected in the control fermentation, while in the stress fermentation they accounted for up to 3 mg/L. These results demonstrate that oxidative stress can play an important role in the final aroma profile; but it is necessary to guarantee adequate yeast growth to obtain the volatile compounds desired. Copyright © 2017 The Institute of Brewing & Distilling     

Application of Plackett–Burman experimental design for investigating the effect of wort amino acids on flavour‐active compounds production during lager yeast fermentation

Aroma‐active higher alcohols and esters are produced intracellularly in the cytosol by fermenting lager yeast cells, which are of major industrial interest because they determine aroma and taste characteristics of the fermented beer. Wort amino acid composition and their utilization by yeast during brewer's wort fermentation influence both the yeast fermentation performance and the flavour profile of the finished product. To better understand the relationship between the yeast cell and wort amino acid composition, Plackett–Burman screening design was applied to measure the changes in nitrogen composition associated with yeast amino acids uptake and flavour formation during fermentation. Here, using an industrial lager brewing strain of Saccharomyces pastorianus , we investigated the effect of amino acid composition on the accumulation of higher alcohols and volatile esters. The objective of this study was to identify the significant amino acids involved in the flavour production during beer fermentation. Our results showed that even though different flavour substances were produced with different amino acid composition in the fermentation experiments, the discrepancies were not related to the total amount of amino acids in the synthetic medium. The most significant effect on higher alcohol production was exercised by the content of glutamic acid, aromatic amino acids and branch chain amino acids. Leucine, valine, glutamic acid, phenylalanine, serine and lysine were identified as important determinants for the formation of esters. The future applications of this information could drastically improve the current regime of selecting malt and adjunct or their formula with desired amino acids in wort. Copyright © 2017 The Institute of Brewing & Distilling     

Free D- and L-Amino Acid Evolution During Sourdough Fermentation and Baking

The evolution of free D- and L-amino acids in sourdoughs started with various lactic acid bacteria (LAB) and yeasts was studied. Lactobacillus brevis subsp. lindneri CB1 and Lactobacillus plantrum DC400 had high proteolytic activity. During sourdough fermentation, Saccharomyces cerevisiae 141 and Saccharomyces exiguus M14 sequentially utilized free amino acids produced by bacterial activity. Due to increased cell yeast autolysis, more S. exiguus M14 inocula caused more free amino acids which were partially utilized by LAB without causing hydrolysis of wheat flour protein. D-alanine, D-glutamic acid and traces of other D-isomers were observed in sourdoughs fermented with L. brevis subsp. lindneri CB1 and S. cerevisiae 141. Free total D- and L-amino acid content decreased by more than 44% after baking the sourdoughs. No abiotic generation of new D-amino acid isomers was detected in the baked sourdoughs.     

Influence of yeast strain,immobilisation and ageing time on the changes of free amino acids and amino acids in peptides in bottle‐fermented sparkling wines obtained from Vitis vinifera cv. Emir

This paper reports the influence of yeast strain, immobilisation and ageing time on the free amino acids and amino acids in peptides of sparkling wine made by the traditional method from the Emir grape variety over a period of 365 days in vintages 2004 and 2005. Four sparkling wines were obtained from the same base wine using yeast strains (both immobilised and free Saccharomyces bayanus and Saccharomyces oviformis). The analysis of variance and cluster revealed that there were significant differences between free amino acids and amino acids in peptides due to ageing time. However, the yeast strain affected most of the free amino acids and amino acids in peptides in only the 2004 vintage. The total amount of free amino acids was higher in the wine made with the S. oviformis compared to S. bayanus. Furthermore, there were no significant differences between the use of immobilised and free yeast in cases of free amino acids and amino acids in peptides.     

Modulation of grape wine flavor via the sequential inoculation of Williopsis saturnus and Saccharomyces cerevisiae

The effects of the inoculum ratio of Williopsis saturnus var. saturnus NCYC22 and Saccharomyces cerevisiae var. bayanus EC-1118 at 1:200 and 1:800 on the chemical and volatile compositions of grape wine were studied in sequential fermentation. The grape juice was first inoculated with Williopsis (W.) saturnus for 9 d; thereafter, Saccharomyces (S.) cerevisiae was inoculated to continue the fermentation until d 19. The cell population of W. saturnus disappeared by d 13, with S. cerevisiae dominating until the end of the fermentation in both inoculum ratios. The changes in yeast count, pH, total soluble solids, sugars, organic acids, and amino acids were similar between the two inoculum ratios. A range of volatile compounds was formed, including alcohols, esters, fatty acids, aldehydes, and terpenes. There were significant differences between both inoculum ratios for medium-chain fatty acids (C8, C10, and C12), ethyl esters of fatty acids of C6, C10, C12, and C14 as well as isoamyl octanoate, while other volatiles were statistically the same.     

Effect of a halophilic aromatic yeast together with Aspergillus oryzae in koji making on the volatile compounds and quality of soy sauce moromi

Koji making is an essential step in the production of high‐quality soy sauce. In this study, we inoculated koji with a halophilic aromatic yeast (Zygosaccharomyces rouxii) and Aspergillus oryzae and investigated the effects on the volatile compounds and quality of soy sauce moromi. Our results suggest that the optimal yeast inoculation was approximately 1.5 × 10⁶ yeast per gram of koji, and excessive yeast greatly decreased A. oryzae enzyme activity. In comparison with the control group, there were no significant changes in the concentrations of amino nitrogen, total titratable acids, total soluble nitrogen and reducing sugar by extending the fermentation time during the first fermentation month, whereas the concentration of glutamate in the yeast‐inoculated sample increased by 46.16% and the arginine concentration decreased by 61.07%. After the second fermentation month, 17, 38 and 38 volatile components were identified by GC–MS in C1, C2 and C5, respectively. Moreover, a large number of volatile components, such as 2‐Methoxy‐4‐vinylphenol, ethyl acetate and 1‐Octen‐3‐ol were found in C2 and C5. And the total contents of volatile compounds in C2 were obviously higher than C5. This work furthers our understanding of the traditional multistrain koji making and provides a new method for enhancing the quality and flavour of soy sauce.     

Ethanol yield and volatile compound content in fermentation of agave must by Kluyveromyces marxianus UMPe-1 comparing with Saccharomyces cerevisiae baker's yeast used in tequila production

In tequila production, fermentation is an important step. Fermentation determines the ethanol productivity and organoleptic properties of the beverage. In this study, a yeast isolated from native residual agave must was identified as Kluyveromyces marxianus UMPe-1 by 26S rRNA sequencing. This yeast was compared with the baker's yeast Saccharomyces cerevisiae Pan1. Our findings demonstrate that the UMPe-1 yeast was able to support the sugar content of agave must and glucose up to 22% (w/v) and tolerated 10% (v/v) ethanol concentration in the medium with 50% cells survival. Pilot and industrial fermentation of agave must tests showed that the K. marxianus UMPe-1 yeast produced ethanol with yields of 94% and 96% with respect to fermentable sugar content (glucose and fructose, constituting 98%). The S. cerevisiae Pan1 baker's yeast, however, which is commonly used in some tequila factories, showed 76% and 70% yield. At the industrial level, UMPe-1 yeast shows a maximum velocity of fermentable sugar consumption of 2.27g·L(-1)·h(-1) and ethanol production of 1.38g·L(-1)·h(-1), providing 58.78g ethanol·L(-1) at 72h fermentation, which corresponds to 96% yield. In addition, the major and minor volatile compounds in the tequila beverage obtained from UMPe-1 yeast were increased. Importantly, 29 volatile compounds were identified, while the beverage obtained from Pan1-yeast contained fewer compounds and in lower concentrations. The results suggest that the K. marxianus UMPe-1 is a suitable yeast for agave must fermentation, showing high ethanol productivity and increased volatile compound content comparing with a S. cerevisiae baker's yeast used in tequila production.     

Ethyl carbamate formation regulated by Saccharomyces cerevisiae ZJU in the processing of Chinese yellow rice wine

Ethyl carbamate (EC) is a probable carcinogen existing in most fermented foods. Throughout traditional fermentation processes, the Chinese fermentation starter plays an important role, but it contains varieties of microorganisms which make inhibiting EC efficiently become a challenge. Therefore, the traditional fermentation starter is substituted with a single yeast strain (Saccharomyces cerevisiae ZJU) to regulate EC catabolism. In this work, S. cerevisiae ZJU can reduce EC formation and the data of EC concentration show that there is 85.6% reduction of EC at most using S. cerevisiae ZJU instead of the traditional fermentation starter. Extracellular urea and citrulline were the leading precursors of EC. The content of amino acids and volatile flavour compounds in the experimental group has no significant influence compared to the natural fermentation. The findings in this work suggest that EC can be regulated by means of the fermentation starters variation.     

Use of spent brewer's yeast in L‐(+) lactic acid fermentation

The application of by‐products from the brewing industry in lactic acid (LA) production was investigated in order to replace expensive nitrogen sources (such as yeast extract) with cheaper and renewable nitrogenous materials such as brewer's yeast (BY). In this study, brewer's spent grain (BSG) hydrolysate was used for L‐(+)‐LA fermentation by Lactobacillus rhamnosus ATCC 7469. The effect of pH control during the fermentation and the addition of various BY contents (5–50 g/L) in BSG hydrolysate on fermentation parameters was evaluated. BY addition significantly increased free amino nitrogen (FAN) concentration (by 25.2% at 5 g/L to 616% at 50 g/L). A strong positive correlation between FAN concentration in the hydrolysate and concentration of L‐(+)‐LA produced was observed (correlation coefficient of 0.913). A high cell viability of L. rhamnosus ATCC 7469 (1.95–3.32 × 10⁹ CFU/mL at the end of fermentation) was achieved in all fermentations with the addition of brewer's yeast. The addition of BY increased L‐(+)‐lactic acid yield and volumetric productivity up to 8.4% (5 g/L) and 48.3% (50 g/L). The highest L‐(+)‐LA yield (89%) and volumetric productivity (0.89 g/L h^?1) were achieved in fermentation of BSG hydrolysate with 50 g/L of BY. © 2019 The Institute of Brewing & Distilling     

Investigation on the elimination of yeasty flavour in yeast extract by mixed culture of lactic acid bacteria and yeast

Yeast extract (YE) is a kind of umami additives used in fermented condiments, it also has special yeasty flavour (YF), which is not widely accepted by some consumers. In this paper, the technology of mixed microbial transformation was applied to eliminate or cover up YF during mixed fermentation. FG10, which is produced by Hubei Angel Yeast Co. Ltd, is a kind of YE that has the distinctive YF and has been applied to the market. It had good sensory evaluation after 6 h with Saccharomycopsis fibuligera and Lactococcus lactis. At 6 h, compared with single fermentation, the number of bacteria of L. lactis was enhanced by 34.28%, and the number of bacteria of S. fibuligera was reduced by 47.93%. The yeasty flavoured substances propionic acid and butyric acid were completely removed. The result of GC-MS showed that acetal, phenyl acetaldehyde, 2,4-dimethylbenzaldehyde, benzaldehyde, isoamyl alcohol, 5-methyl-2-furan methanol, phenylethanol were generated or increased. Furthermore, the amino acids of YE were significantly different after fermentation by different microorganisms (P < 0.05), The flavour compounds and amino acids in YE before and after fermentation had closely correlation (r = 0.94), the consumption of amino acids in mixed fermentation promoted the variety of corresponding flavour compounds, which boosted the formation of free YF YE. In addition, taste substances lactic acid generated 6.27 ± 0.115 g L⁻¹ and nucleotides improved 15.05%, which were beneficial to natural YF free seasoning with YE as raw material.     

Release of nitrogen compounds to the extracellular medium by three strains of Saccharomyces cerevisiae during induced autolysis in a model wine system.

To detect differences among three strains of Saccharomyces cerevisiae used in the manufacture of sparkling wines and to study the changes in nitrogen compounds during autolysis, a model wine system was used. Significant differences were observed between the mean values of the autolytic capacity of the three strains. The amount of nitrogen (total, protein, peptide and amino) present in the autolysates and the concentration of most free amino acids was significantly affected by the strain. These findings suggest that the strain of yeast used in the manufacture of sparkling wines can play an important role in the aging process and can affect final composition.