Zygosaccharomyces Rouxii Research Articles

The revenge of Zygosaccharomyces yeasts in food biotechnology and applied microbiology.

Non-conventional yeasts refer to a huge and still poorly explored group of species alternative to the well-known model organism Saccharomyces cerevisiae. Among them, Zygosaccharomyces rouxii and the sister species Zygosaccharomyces bailii are infamous for spoiling food and beverages even in presence of several food preservatives. On the other hand, their capability to cope with a wide range of process conditions makes these yeasts very attractive factories (the so-called "ZygoFactories") for bio-converting substrates poorly permissive for the growth of other species. In balsamic vinegar Z. rouxii is the main yeast responsible for converting highly concentrated sugars into ethanol, with a preference for fructose over glucose (a trait called fructophily). Z. rouxii has also attracted much attention for the ability to release important flavor compounds, such as fusel alcohols and the derivatives of 4-hydroxyfuranone, which markedly contribute to fragrant and smoky aroma in soy sauce. While Z. rouxii was successfully proposed in brewing for producing low ethanol beer, Z. bailii is promising for lactic acid and bioethanol production. Recently, several research efforts exploited omics tools to pinpoint the genetic bases of distinctive traits in "ZygoFactories", like fructophily, tolerance to high concentrations of sugars, lactic acid and salt. Here, I provided an overview of Zygosaccharomyces industrially relevant phenotypes and summarized the most recent findings in disclosing their genetic bases. I suggest that the increasing number of genomes available for Z. rouxii and other Zygosaccharomyces relatives, combined with recently developed genetic engineering toolkits, will boost the applications of these yeasts in biotechnology and applied microbiology.

Non-conventional yeasts from fermented honey by-products: Focus on Hanseniaspora uvarum strains for craft beer production

The increasing interest in novel beer productions focused on non-Saccharomyces yeasts in order to pursue their potential in generating groundbreaking sensory profiles. Traditional fermented beverages represent an important source of yeast strains which could express interesting features during brewing. A total of 404 yeasts were isolated from fermented honey by-products and identified as Saccharomyces cerevisiae, Wickerhamomyces anomalus, Zygosaccharomyces bailii, Zygosaccharomyces rouxii and Hanseniaspora uvarum. Five H. uvarum strains were screened for their brewing capability. Interestingly, Hanseniaspora uvarum strains showed growth in presence of ethanol and hop and a more rapid growth than the control strain S. cerevisiae US-05. Even though all strains showed a very low fermentation power, their concentrations ranged between 7 and 8 Log cycles during fermentation. The statistical analyses showed significant differences among the strains and underlined the ability of YGA2 and YGA34 to grow rapidly in presence of ethanol and hop. The strain YGA34 showed the best technological properties and was selected for beer production. Its presence in mixed- and sequential-culture fermentations with US-05 did not influence attenuation and ethanol concentration but had a significant impact on glycerol and acetic acid concentrations, with a higher sensory complexity and intensity, representing promising co-starters during craft beer production.

Toward application of biocontrol to inhibit wine spoilage yeasts: The use of statistical designs for screening and optimisation

Spoilage yeasts generate considerable economic losses in the wine industry, and although sulphur dioxide (SO2) is traditionally used for control, its use has become controversial because of its negative effects on health. Biocontrol has emerged as a partial alternative to SO2, and most research has focused on the selection of biocontrol yeasts and/or the mechanisms involved, while little research has been directed to the environmental conditions that make biocontrol effective for application. When there are two or more interacting yeasts, the physicochemical factors that affect their antagonism are many and therefore the application of biocontrol is complex. To reduce SO2, the present study aimed to elucidate biocontrol mechanisms of two yeast interactions and to establish optimal physicochemical conditions for biocontrol of the spoilage yeast during grape must fermentation. Through the use of statistical design, it was possible to find relevant physicochemical factors and optimise them. Wickerhamomyces anomalus “BWa156” developed an active supernatant against ZygoSaccharomyces rouxii “BZr6” while supernatant from Metschnikowia pulcherrima “BMp29” was ineffective. In mixed must fermentations, the first interaction (BWa156 vs. BZr6) showed fewer physicochemical factors impacting biocontrol compared to the second interaction (BMp29 vs. BZr6). However, the fewer factors of the first interaction had a stronger effect on the decline in the spoilage population. Validations showed that the optimal conditions for biocontrol with the first interaction could be predicted. Analysis of the results with BWa156 vs. BZr6 and BMp29 vs. BZr6 suggests that the first interaction is a competition that includes a killer toxin, while the second interaction involves competition for iron resources. Response surface methodology (RSM) allowed a reduction in the number of experiments and permitted to find the optimal biocontrol conditions (SO2: 0 mg mL-1; pH: 3.7; Reducing sugars: 23 °Brix) for the interaction between BWa156 and BZr6.

Ester synthesis mechanism and activity by Bacillus licheniformis, Candida etchellsii, and Zygosaccharomyces rouxii isolated from Chinese horse bean chili paste.

Esters are indispensable aroma compounds and contribute significantly to the fruity aromas in fermented condiments. The ester synthesis activity and pathways of Bacillus licheniformis, Candida etchellsii, and Zygosaccharomyces rouxii, isolated from Chinese horse bean chili-paste (CHCP), were investigated. Chemical buffer models containing esterification and alcoholysis systems inoculated with extracellular extracts of these three strains were established. The ester synthesis activity of C. etchellsii was stronger than that of the other two strains. Zygosaccharomyces rouxii could synthesize acetate esters via esterification, whereas the biosynthesis pathways of B. licheniformis and C. etchellsii were esterification and alcoholysis. Esterification exhibited relatively high activity at pH4, whereas alcoholysis activity improved with an increase in the pH from 4 to 8. Candida etchellsii could synthesize C6 -C8 of acetate esters, and its activity improved with the number of alcohol carbon atoms. These three strains could synthesize C10 -C18 of ethyl esters. Their ethyl ester synthesis activity decreased with the aliphatic acid carbon number. Candida etchellsii has the potential to be used in CHCP fermentation to accumulate esters and improve flavor compared with the other two strains. This research is helpful in explaining the mechanism of ester synthesis in fermented condiments. © 2021 Society of Chemical Industry.

Development of a defined autochthonous starter through dissecting the seasonal microbiome of broad bean paste

Bean-based fermentation foods are usually ripened in open environment, which would lead to inconsistencies in flavor and quality between batches. The physicochemical metabolism and microbial community of seasonal broad bean paste (BBP) were compared to distinguish discriminant metabolites and unique taxa, as well as their specific reasons for different flavor and quality in this study. Here, we found that environmental variables led to the seasonal distribution of microbiota, and differential microorganisms further contributed to the inconsistency of flavor quality, in which Lactobacillales was responsible for the higher titratable acid and amino acid nitrogen concentration in winter pei, while Saccharomycetales benefited the formation of volatile flavor substances in autumn pei. Additionally, we compared the effect of different combinations of Lactobacillales with Zygosaccharomyces rouxii on the quality of BBP, and found that W. confusa was more suitable for BBP fermentation rather than T. halophilus in terms of sensory characteristics and physicochemical metabolites.

Effect of Tetragenococcus halophilus, Zygosaccharomyces rouxii, and Torulopsis versatilis addition sequence on soy sauce fermentation

The aim of this study was to investigate the effect that the sequences of ingredient addition had on soy sauce fermentation. Five strategies were assessed for the inoculation of Tetragenococcus halophilus, Zygosaccharomyces rouxii, and Torulopsis versatilis. The results of physicochemical analysis and levels of flavor compounds demonstrated that the best strategy for soy sauce fermentation was to inoculate T. halophilus (2 × 105 CFU/mL) on the 15th day, Z. rouxii (1.0 × 106 CFU/mL) on the 30th day, and T. versatilis (1.0 × 106 CFU/mL) on the 45th day; this enhanced its fruity, sauce, alcoholic, and caramel-like flavors by 64.3%, 22.7%, 43.1%, and 36.2%, respectively, and saline taste by 64.3%. Spearman correlation analysis revealed the content of ethanol positively correlated with acetic acid (r = 0.82) and succinic acid (r = 0.80), while the level of formic acid negatively correlated with ethyl phenylacetate (r = −0.88). In summary, these results provided fundamental evidence for strain addition sequence for producing flavor-enhanced soy sauce with potential application in the future.

Use of non-Saccharomyces yeasts in cider fermentation: Importance of the nutrients addition to obtain an efficient fermentation

The isolation of autochthonous yeast species presents a good strategy to select new microorganisms for developing an adequate inoculum to carry out fermentations and generate representative products of the cider production zone. However, non-Saccharomyces yeasts have been considered to have low capacity to carry out a complete fermentation as Saccharomyces cerevisiae. In this work, five autochthonous yeasts from a cider fermentation process were isolated and identified as Saccharomyces cerevisiae, Kluyveromyces marxianus, Pichia membranaefaciens, P. kluyveri and Zygosaccharomyces rouxii. A series of fermentations were developed at laboratory level, using each species individually and it was observed that only S. cerevisiae was able to finish the process. K. marxianus consumed less than 50% of the sugars; P. kluyveri and Z. rouxii consumed less than 70% and P. membranaefaciens consumed more than 90% but the yield (ethanol produced for sugar consumed (YP/S)) was 0.39. Nevertheless, the addition of magnesium, zinc and nitrogen increased the fermentative capacity of almost all species: K. marxianus, Z. rouxii and P. kluyveri, showed an increase in ethanol production when nutrients were added, obtaining more than 80 g/L of ethanol, and showing that those nutrients are necessary to complete the fermentation. This work describes the potential use of different non-Saccharomyces species to carry out fermentation of apple juice and highlights the importance of certain nutrients to enable an efficient alcoholic fermentation and the generation of desirable volatile compounds for cider production.

Metabarcoding analysis and fermentation performance of the dominant fungal microbiota associated with the Algerian traditional date product "Btana".

The study highlights the fungal diversity of the traditional Algerian date's product "Btana" prepared with direct Btana method (DBM) and undirect Btana method (UBM). Btana fungal populations were analyzed through 28S metabarcoding. Data treatment resulted in 122,997 reads representing three Phyla in which 76% reads (46 OTUs) belong to Ascomycota phylum. Zygosaccharomyces rouxii was the most prevailed species accounting for 35.40% of the total population. Similarity percentage analysis revealed a low level of resemblance in species in each of the two Btana types (DBM: 17.26%, UBM: 16.87). According to HPLC analysis, lactate was detected in nine samples within a range of 0.87-23.06 g/100g. Culture plating and subsequent D1/D2 domain of 28s DNA analysis showed the prevalence of Z. rouxii. Fermentation of non-renewed date medium revealed a high ethanol production (21.31 ± 2.89 g/100g) by Lachancea thermotolerans and 5.87 g/100g of lactates by Kluyveromyces delphensis. Enzymatic assay revealed a high esterase (C4) and naphtol-AS-BI-phosphohydrolase activity by L. thermotolerans, K. delphensis, and Pichia subpelliculosa, while a high level of α-fucosidase was recorded for L. thermotolerans and P. subpelliculosa. The current results demonstrated that the traditional date product Btana is a promising source for yeasts useful in production of value-added products like bioethanol and lactic acid using low-income date cultivars.

Effects of microwave pasteurization on the thermal resistance of Zygosaccharomyces rouxii and on the physicochemical properties of organic intermediate moisture raisin

In this study, organic intermediate moisture raisin was pasteurized with microwave and conventional pasteurization methods. The thermal resistance (D and z values) of the target yeast, Zygosaccharomyces rouxii, was determined for both methods at different temperatures 60°C, 70°C, and 80°C and at different time intervals. The effect of pasteurization methods on the quality properties of raisin samples was investigated. The D values of Z. rouxii were found as 6.50 ± 0.49, 2.96 ± 0.02, 1.91 ± 0.01 min for conventional treatment and 3.46 ± 0.31, 2.02 ± 0.14 and 1 ± 0.01 min for microwave treatment at 60℃, 70℃, and 80℃, respectively. F8537 values for 5-log reduction and come-up time significantly decreased after microwave pasteurization. Brix, acidity, and color values were preserved better by microwave treatment. Hydroxymethylfurfural content of the microwave pasteurized samples was found 42% less than the samples pasteurized conventionally. Microwave pasteurization can be alternatively used for the conventional method for raisin improving quality with less heating time. Practical applications Microwave heating needs less come-up time rather than conventional pasteurization. Decreasing process time provides advantageous via the costs and energy saving so organic intermediate moisture fruits that are perishable can be safely pasteurized by a microwave method for longer shelf life in the industry. Z. rouxii, which may be found in high sugar content foods, can be inactivated to the target log that provides the fruit safe by using this method. In addition, quality properties, such as color, which is effective on the consumer preference also, protected better by the effect of volumetric heating.

Vinification without Saccharomyces: Interacting Osmotolerant and "Spoilage" Yeast Communities in Fermenting and Ageing Botrytised High-Sugar Wines (Tokaj Essence).

The conversion of grape juice to wine starts with complex yeast communities consisting of strains that have colonised the harvested grape and/or reside in the winery environment. As the conditions in the fermenting juice gradually become inhibitory for most species, they are rapidly overgrown by the more adaptable Saccharomyces strains, which then complete the fermentation. However, there are environmental factors that even Saccharomyces cannot cope with. We show that when the sugar content is extremely high, osmotolerant yeasts, usually considered as “spoilage yeasts“, ferment the must. The examination of the yeast biota of 22 botrytised Tokaj Essence wines of sugar concentrations ranging from 365 to 752 g∙L−1 identified the osmotolerant Zygosaccharomyces rouxii, Candida (Starmerella) lactis-condensi and Candida zemplinina (Starmerella bacillaris) as the dominating species. Ten additional species, mostly known as osmotolerant spoilage yeasts or biofilm-producing yeasts, were detected as minor components of the populations. The high phenotypical and molecular (karyotype, mtDNA restriction fragment length polymorphism (RFLP) and microsatellite-primed PCR (MSP-PCR)) diversity of the conspecific strains indicated that diverse clones of the species coexisted in the wines. Genetic segregation of certain clones and interactions (antagonism and crossfeeding) of the species also appeared to shape the fermenting yeast biota.

Levaduras aisladas de mieles como antagonistas de mohos patógenos de cultivos

Se buscó el aislamiento de levaduras de muestras de miel de la Provincia de Jujuy, Argentina, con el objetivo de utilizarlas in vitro como antagonistas de mohos patógenos pre- y poscosecha, de diferentes cultivos de interés, como, cítricos, maíz, maní y caña de azúcar. Se utilizó miel, ya que la misma puede actuar como medio selectivo de cepas inocuas y tolerantes a distintos estrés abióticos, como bajo pH, elevada presión osmótica, presencia de fitoquímicos, entre otros. Se logró aislar un total de 15 levaduras de 25 muestras de mieles. Las levaduras fueron identificadas como: Candida parapsilosis, Zygosaccharomyces baili, Zygosaccharomyces mellis, Zygosaccharomyces rouxii, Tausonia pullulans, Lachancea thermotolerans, Lachancea fermentati, Torulaspora delbrueckii y Saccharomyces cerevisiae. Todos los géneros se encontraron descriptos como presentes en la miel, provenientes, ya sea, de fuentes primarias de contaminación o del ambiente circundante. De los aislamientos obtenidos se probó el antagonismo in vitro, por disminución del crecimiento micelial de los mohos: Penicillium italicum, P. ulaiense, Aspergillus parasiticus y Fusarium sacchari. Lachancea thermotolerans, fue el único aislamiento que presentó antagonismo hacia todos los mohos ensayados. Candida parapsilosis manifestó antagonismo hacia P. italicum; Lachancea fermentati hacia P. ulaiense; Z. mellis frente P. ulaiense y uno de los aislamientos de T. delbrueckii frente a P. italicum y P. ulaiense. Se podrían utilizar las levaduras antagonistas en futuros ensayos in vivo, en vistas al diseño de un biofungicida activo contra mohos patógenos de cultivos, en la etapa de producción a campo o durante el almacenamiento.

Heat preadaptation improved the ability of Zygosaccharomyces rouxii to salt stress: a combined physiological and transcriptomic analysis.

Zygosaccharomyces rouxii plays important roles in the brewing process of fermented foods such as soy sauce, where salt stress is a frequently encountered condition. In this study, effect of heat preadaptation on salt tolerance of Z. rouxii and the protective mechanisms underlying heat preadaptation were investigated based on physiological and transcriptomic analyses. Results showed that cells subjected to heat preadaptation (37°C, 90min) prior to salt stress aroused many physiological responses, including maintaining cell surface smooth and intracellular pH level, increasing Na+/K+-ATPase activity. Cells subjected to heat preadaptation increased the amounts of unsaturated fatty acids (palmitoleic C16:1, oleic C18:1, linoleic C18:2) and decreased the amounts of saturated fatty acids (palmitic C16:0, stearic C18:0) which caused the unsaturation degree (unsaturated/saturated = U/S ratio) increased by 2.4 times when compared with cells without preadaptation under salt stress. Besides, salt stress led to increase in contents of 5 amino acids (valine, proline, threonine, glycine, and tyrosine) and decrease of 2 amino acids (serine and lysine). When comparing the cells pre-exposed to heat preadaptation followed by challenged with salt stress and the cells without preadaptation under salt stress, the serine, threonine, and lysine contents increased significantly. RNA sequencing revealed that the metabolic level of glycolysis by Z. rouxii was weakened, while the metabolic levels of the pentose phosphate pathway and the riboflavin were enhanced in cells during heat preadaptation. Results presented in this study may contribute to understand the bases of adaptive responses in Z. rouxii and rationalize its exploitation in industrial processes.Key points• Heat preadaptation can improve high salinity tolerance of Z. rouxii.• Combined physiological and transcriptomic analyses of heat preadaptation mechanisms.• Provide theoretical support for the application of Z. rouxii.

Effect of aroma-producing yeasts in high-salt liquid-state fermentation soy sauce and the biosynthesis pathways of the dominant esters

Fermentation with excellent aroma-producing yeasts can enhance the flavour of soy sauce. In this work, Millerozyma farinosa CS2.23, Zygosaccharomyces rouxii CS2.42, and Candida parapsilosis CS2.53 were added to the high-salt liquid-state moromi to promote soy sauce fermentation. All three yeasts improved the TE of soy sauce, the highest of which reached 1.03 g/L with added CS2.42. Other quality indexes of soy sauce, including RS, TA, and AN, were not greatly affected. The volatile esters of soy sauce added to the three yeasts increased by 108.85%, 166.71%, and 113.61% compared with the control through GC-MS analysis. Obviously, CS2.42 had an excellent ability to produce esters. Studying the biosynthesis pathway of esters, CS2.42 has the best esterification ability, while CS2.53 has the advantage of alcoholysis ability. The exploration of the biosynthetic pathway of acetate and ethyl esters has laid a foundation for regulating esters in soy sauce fermentation.

Zygosaccharomyces rouxii, an Aromatic Yeast Isolated From Chili Sauce, Is Able to Biosynthesize 2-Phenylethanol via the Shikimate or Ehrlich Pathways.

We isolated an aromatic strain of yeast (M2013310) from chili sauce. Assembly, annotation, and phylogenetic analysis based on genome sequencing, identified M2013310 as an allodiploid yeast that was closely related to Zygosaccharomyces rouxii. During fermentation, M2013310, produced an aromatic alcohol with a rose-honey scent; gas chromatography tandem mass spectrometry identified this alcohol as 2-phenylethanol. The concentration of 2-phenylethanol reached 3.8 mg/L, 1.79 g/L, and 3.58 g/L, in M3 (NH4+), M3 (NH4+ + Phe), and M3 (Phe) culture media, after 72 h of fermentation, respectively. The mRNA expression levels of ARO8 encoding aromatic aminotransferases I and ARO10 encoding phenylpyruvate decarboxylase by M2013310 in M3 (Phe) were the lowest of the three different forms of media tested. These results indicated that M2013310 can synthesize 2-phenylethanol via the Shikimate or Ehrlich pathways and the production of 2-phenylethanol may be significantly improved by the over-expression of these two genes. Our research identified a promising strain of yeast (M2013310) that could be used to improve the production of 2-phenylethanol.

Genomic scanning of the promoter sequence in osmo/halo-tolerance related QTLs in Zygosaccharomyces rouxii

Zygosaccharomyces rouxii, an osmotic stress-tolerant yeast species is conventionally used in high sugar/saline food supplements and in beverage industries. The genetic basis of phenotypic and fundamental diversity of the species is less understood. In this study, we scanned the promoter region of 50 osmo/halo-tolerant genes in yeast species with special reference to Zygosaccharomyces rouxiiin order to understand their distinct regulation during osmo or halo-stress condition. In their promoter region, we have focused on the stress regulatory elements (STREs), repetitive elements, and the transcription factor (TF) binding sites of the genes. These motifs are known to play a key role in the regulation of the transcription of osmo-responsive genes. On analyzing the promoter, we came to know that out of 50 genes analyzed, 18 genes possess STREs, 26 genes harbor tandem repeat sequences and 18 genes have transcription factor binding sites in their promoter region. Herein, we aim to describe how promoters of the osmo/halotolerant genes that possess these STRE elements, tandem repeats, and TF sites are expected to display considerable divergence in the transcriptional activity of the associated gene. We also aim to show that variations in STRE elements, tandem repeat number/length and TFs binding sites alters local nucleosome positioning resulting into differential gene expression. The understanding that would generate from the current study is expected to highly useful in exploitation of yeast in industrial set ups. Acknowledgment of these factors can lead to the development of a better understanding of the physiological mechanisms of tolerance against osmo/halo-stress in the genus Zygosaccharomyces.

Molecular mechanism of the response of Zygosaccharomyces rouxii to D-fructose stress by the glutathione metabolism pathway.

Zygosaccharomyces rouxii produces high levels of 4-hydroxy-2,5-dimethyl-3(2H)-furanone in YPD medium supplemented with 120g/L D-fructose and 180g/L NaCl after 5 d. D-fructose has a stress effect on Z. rouxii, and GSH-Px is a main enzyme involved in the defense of Z. rouxii against oxygen stress according to our previous report. In order to further explore the molecular mechanism of the glutathione metabolism pathway in Z. rouxii in response to D-fructose stress, changes in the expression of genes and proteins involved in the synthesis of glutathione precursor amino acids and enzymes were observed. In addition, changes in the intermediates related to glutathione synthesis in Z. rouxii were reported. The results indicated that some gene-encoding enzymes involved in the glutamate, cysteine and glycine biosynthesis pathways and key genes involved in glutathione synthesis were upregulated. The expression levels of other genes, except SHMT, were consistent with the qRT-PCR results. The contents of γ-glutamylcysteine and glutathione amide in the D-fructose group were higher than those in the control group. In the D-fructose stress groups, the metabolic flux towards glutathione synthesis was increased. These results might provide more in-depth and detailed theoretical support for the oxidative stress defense mechanism of Z. rouxii under D-fructose stress.

A Bottom-Up Approach To Develop a Synthetic Microbial Community Model: Application for Efficient Reduced-Salt Broad Bean Paste Fermentation.

Humans have used high salinity for the production of bean-based fermented foods over thousands of years. Although high salinity can inhibit the growth of harmful microbes and select functional microbiota in an open environment, it also affects fermentation efficiency of bean-based fermented foods and has a negative impact on people's health. Therefore, it is imperative to develop novel defined starter cultures for reduced-salt fermentation in a sterile environment. Here, we explored the microbial assembly and function in the fermentation of traditional Chinese broad bean paste with 12% salinity. The results revealed that the salinity and microbial interactions together drove the dynamic of community and pointed out that five dominant genera (Staphylococcus, Bacillus, Weissella, Aspergillus, and Zygosaccharomyces) may play different key roles in different fermentation stages. Then, core species were isolated from broad bean paste, and their salinity tolerance, interactions, and metabolic characteristics were evaluated. The results provided an opportunity to validate in situ predictions through in vitro dissection of microbial assembly and function. Last, we reconstructed the synthetic microbial community with five strains (Aspergillus oryzae, Bacillus subtilis, Staphylococcus gallinarum, Weissella confusa, and Zygosaccharomyces rouxii) under different salinities and realized efficient fermentation of broad bean paste for 6 weeks in a sterile environment with 6% salinity. In general, this work provided a bottom-up approach for the development of a simplified microbial community model with desired functions to improve the fermentation efficiency of bean-based fermented foods by deconstructing and reconstructing the microbial structure and function.IMPORTANCE Humans have mastered high-salinity fermentation techniques for bean-based fermented product preparation over thousands of years. High salinity was used to select the functional microbiota and conducted food fermentation production with unique flavor. Although a high-salinity environment is beneficial for suppressing harmful microbes in the open fermentation environment, the fermentation efficiency of functional microbes is partially inhibited. Therefore, application of defined starter cultures for reduced-salt fermentation in a sterile environment is an alternative approach to improve the fermentation efficiency of bean-based fermented foods and guide the transformation of traditional industry. However, the assembly and function of self-organized microbiota in an open fermentation environment are still unclear. This study provides a comprehensive understanding of microbial function and the mechanism of community succession in a high-salinity environment during the fermentation of broad bean paste so as to reconstruct the microbial community and realize efficient fermentation of broad bean paste in a sterile environment.

Identification of SFL1 as a positive regulator for flor formation in Zygosaccharomyces rouxii.

Some wild Zygosaccharomyces rouxii impair the quality of soy sauce through the generation of unpleasant odors induced by the formation of flor. Flor formation in Z. rouxii depends on the expression of the FLO11D gene, which is a homolog of the FLO11 gene that encodes a cell surface protein in Saccharomyces cerevisiae. FLO11 expression in S. cerevisiae is regulated by multiple pathways. To investigate the regulation of FLO11D expression in Z. rouxii, we created 13 gene knockout mutants (STE12, TEC1, HOG1, MSS11, FLO8, MSN1, MSN2/4, SKO1, TUP1, CYC8, YAK1, MIG1, and SFL1) related to those pathways and examined whether these mutants form flor. Unexpectedly, SFL1 knockout mutant could only form a very weak flor due to decreased FLO11D expression, suggesting that SFL1 acts as a potential activator of flor formation through FLO11D expression. This result is in contrast to S. cerevisiae SFL1, which acts as a repressor of FLO11 expression.

Accelerating aroma formation of raw soy sauce using low intensity sonication

Sonication was applied to accelerate aroma formation and shorten fermentation time of soy sauce. Effects of sonication at 68 kHz on the aroma and aroma-producing Zygosaccharomyces rouxii and Tetragenococcus halophilus in moromis were investigated using sensory evaluation, SPME–GC–olfactometry/MS, viable cell counting and scanning electron microscopy. The sensory scores of caramel-like, fruity, alcoholic, floral, malty, smoky, sour and overall aroma in sonicated moromis were enhanced by 23.4%, 23.2%, 13.6%, 12.8%, 7.6%, 6.3%, 5.6% and 14.4%, respectively. Sensory scores of samples fermented for 90–180 days were higher than those of controls fermented for 180 days, suggesting that sonication could reduce fermentation time by 90 days. Thirty-four aroma-active compounds were detected from 85 volatile compounds in soy sauces. Sonication accelerated and elevated the formation of aroma compounds by chemical reactions. It also markedly increased the reproduction and cell permeability of both microorganisms in moromis, which favored the formation of aroma compounds by both strains.

Assessment of different antimicrobials to inhibit the growth of Zygosaccharomyces rouxii cocktail in concentrated apple juice

Zygosaccharomyces rouxii represents the main spoilage cause of concentrated apple juice, leading to waste of products or recalls. Essential oils components derived from plants have been found to present antimicrobial activities against various microbes. However, few work has been reported about their antimicrobial activities against Z. rouxii in concentrated apple juice. In this work, reparameterized Gompertz equation was used to evaluate the antimicrobial activities of different antimicrobials to inhibit the growth of a Z. rouxii cocktail (6.3 lg colony forming units/mL) composed of six strains isolated from concentrated apple juice and two strains from honey and grape must. The obtained mathematical models presented that thymol, carvacrol and trans-cinnamaldehyde were the promising options to inhibit Z. rouxii in 30 oBrix apple juice, reaching a maximal decrease on yeast growth of around 99.65 ± 0.61%. Whereas other antimicrobials showed lower antimicrobial activities with a maximal growth decrease of ranging from 67.13 ± 3.62% to 13.38 ± 1.16%. Additionally, the sensorial characteristics were not affected when the antimicrobials assayed were applied at the effective concentrations in commercial apple juice product. This work provided a theoretical feasibility that thymol, carvacrol and trans-cinnamaldehyde could be applied as natural preservatives for the control of Z. rouxii-related spoilage in fruit juice industry.