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Results for “spontaneous fermentation”

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PubMed1989

[Yeasts associated with spontaneous fermentation processes in wines from Ribeiro. Analysis of homo/heterothallism and the killer system of S. cerevisiae strains].

A taxonomic analysis has been carried out concerning the different yeast species involved in spontaneous fermentations of Ribeiro wine. The results revealed the presence of 15 species belonging to 7 genera: Saccharomyces, Torulaspora, Dekkera, Saccharomycodes, Debaryomyces, Kloeckera and Candida. Within the genus Candida a high number of species was detected. The 28 isolated strains of S. cerevisiae showed with some exceptions, a remarkable sporulation ability and viability of the meiotic products. All the strains were homothallic. Among all S. cerevisiae strains, 6 exhibited killer K2 activity with higher isolation percentages in intermediate fermentations than in late fermentations.

Fermentation↗

PubMed2006

Spontaneously fermented millet product as a natural probiotic treatment for diarrhoea in young children: an intervention study in Northern Ghana.

Indigenous lactic acid fermented foods may have potential as probiotic treatment for diarrhoea, due to high levels of lactic acid bacteria. In this study the effect of a millet drink, spontaneously fermented by lactic acid bacteria, as a therapeutic agent among Ghanaian children with diarrhoea, was assessed. Children below 5 years of age coming to Northern Ghana health clinics for treatment of diarrhoea were randomised to two groups. Children of both groups received treatment for diarrhoea given at the local clinic. The intervention group in addition received up to 300 ml fermented millet drink (KSW) daily for 5 days after enrolment. The clinical outcome of diarrhoea and reported well-being were registered every day for the 5-day intervention and again 14 days after diagnosis. Among 184 children (mean age 17.4, standard deviation 11.3 months) included, no effects of the intervention were found with respect to stool frequency, stool consistency and duration of diarrhoea. However, KSW was associated with greater reported well-being 14 days after the start of the intervention (P=0.02). The fact that no effect of KSW on diarrhoea was observed could be because many children had a mild form of diarrhoea, and many were treated with antibiotics. Either this could have affected the lactic acid bacteria, or the lactic acid bacteria in KSW had no probiotic effects. It is speculated that the effect after two weeks could be due to a preventing effect of KSW on antibiotic-associated diarrhoea which could help reducing persistent diarrhoea.

Anti-Bacterial Agents↗

PubMed2003

Identification of lactic acid bacteria from spontaneous fermentation of 'Almagro' eggplants by SDS-PAGE whole cell protein fingerprinting.

In order to complete the previously performed phenotypic identification [Sánchez et al., 2000. Int. J. Food Microbiol. 59 (2000) 9], whole cell protein patterns obtained using Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE) of 149 lactic acid bacteria (LAB) strains, isolated from 'Almagro' eggplants spontaneous fermentation, were analysed. Phenotypic identification of those strains had revealed the presence of the species Lactobacillus (Lb) plantarum, Lb. pentosus, Lb. brevis biotype 2, Lb. brevis biotype 3, Lb. fermentum, Lactobacillus spp. and Aerococcus viridans. The SDS-PAGE technique generated complex and stable patterns that were easy to interpret and compare with those of the 17 reference strains used in this study. After numerical analysis of the resulting electrophoretic protein patterns, six well-delineated clusters were discriminated that, with some exceptions, correlated well with the different groups phenotypically found. Only two strains showed unique protein patterns without meaningful homology to any of the reference strains used for this study and remained unidentified.

Cluster Analysis↗

PubMedDate not supplied

Dimorphic fungi isolated from spontaneously fermented juice of soursop, Annona Muricata L.

Two new fungi isolated from fermenting juice of soursop Annona Muricata L., exhibited dimorphism. Aerobic hyphae were coenocytic bearing sporangia while vegetative filaments were septate. Growth in broth was in discrete units. Glucose--yeast extract-peptone broth inoculated with sporangiospores of strain C12 induced arthroconidiospores but it was yeast cells, yeastlike cells, pseudohyphae and pseudomycelia bearing blastospores when strain C13 was inoculated. On the other hand, soursop extract induced arthrospores and yeast cells, yeast like cells and blastospore-bearing pseudomycelia respectively with strain C12 and strain C13 inoculation. Physiological characteristics were distinct. Strain C12 fermented soluble starch and raffinose completely while strain C13 proved weak for both sugars but complete in glucose utilization. The two dimorphic strains along with Saccaharomyces latis were negative in inulin fermentation. The three strains assimilated all carbon and nitrogen sources tested and grew at 37 degrees C. Based on cultural, morphological and biochemical differences, a tentative genus, Dimorphomyces was created for the dimorphic strains. D. diastaticus strain C12 and D. pleomorphis strain C13 were thought to initiate spontaneous fermentation which was brought to completion along with S. latis strain C14.

Culture Media↗

PubMed1997

Isolation and molecular characterization of high-performance cellobiose-fermenting spontaneous mutants of ethanologenic Escherichia coli KO11 containing the Klebsiella oxytoca casAB operon.

Escherichia coli KO11 was previously constructed to produce ethanol from acid hydrolysates of hemicellulose (pentoses and hexoses) by the chromosomal integration of Zymomonas mobilis genes encoding pyruvate decarboxylase (pdc) and alcohol dehydrogenase (adhB). Klebsiella oxytoca P2 was constructed in an analogous fashion for the simultaneous saccharification and fermentation of cellulose and contains PTS enzymes for cellobiose. In this study, KO11 was further engineered for the fermentation of cellulose by adding the K. oxytoca casAB genes encoding Enzyme IIcellobiose and phospho-beta-glucosidase. Although the two K. oxytoca genes were well expressed in cloning hosts such as DH5 alpha, both were expressed poorly in E. coli KO11, a derivative of E. coli B. Spontaneous mutants which exhibited more than 15-fold-higher specific activities for cellobiose metabolism were isolated. The mutations of these mutants resided in the plasmid rather than the host. Three mutants were characterized by sequence analysis. All contained similar internal deletions which eliminated the casAB promoter and operator regions and placed the lacZ Shine-Dalgarno region immediately upstream from the casA Shine-Dalgarno region. KO11 harboring mutant plasmids (pLOI1908, pLOI1909, or pLOI1910) rapidly fermented cellobiose to ethanol, and the yield was more than 90% of the theoretical yield. Two of these strains were used with commercial cellulase to ferment mixed-waste office paper to ethanol.

Base Sequence↗

PubMed2006

Yeast species associated with the spontaneous fermentation of cider.

This paper reports the influence of cider-making technology (pneumatic and traditional pressing) on the dynamics of wild yeast populations. Yeast colonies isolated from apple juice before and throughout fermentation at a cider cellar of Asturias (Spain), during two consecutive years were studied. The yeast strains were identified by restriction fragment length polymorphism analysis of the 5.8S rRNA gene and the two flanking internal transcribed sequences (ITS). The musts obtained by pneumatic pressing were dominated by non-Saccharomyces yeasts (Hanseniaspora genus and Metschnikowia pulcherrima) whereas in the apple juices obtained by traditional pressing Saccharomyces together with non-Saccharomyces, were always present. The species Saccharomyces present were S. cerevisiae and S. bayanus. Apparently S. bayanus, was the predominant species at the beginning and the middle fermentation steps of the fermentation process, reaching a percentage of isolation between 33% and 41%, whereas S. cerevisiae took over the process in the final stages of fermentation. During the 2001 harvest, with independence of cider-making technology, the species Hanseniaspora valbyensis was always isolated at the end of fermentations.

Base Sequence↗

PubMed2001

Yeast population dynamics in spontaneous fermentations: comparison between two different wine-producing areas over a period of three years.

Yeast ecology, biogeography and biodiversity are important and interesting topics of research. The population dynamics of yeasts in several cellars of two Spanish wine-producing regions was analysed for three consecutive years (1996 to 1998). No yeast starter cultures had been used in these wineries which therefore provided an ideal winemaking environment to investigate the dynamics of grape-related indigenous yeast populations. Non-Saccharomyces yeast species were identified by RFLPs of their rDNA, while Saccharomyces species and strains were identified by RFLPs of their mtDNA. This study confirmed the findings of other reports that non-Saccharomyces species were limited to the early stages of fermentation whilst Saccharomyces dominated towards the end of the alcoholic fermentation. However, significant differences were found with previous studies, such as the survival of non-Saccharomyces species in stages with high alcohol content and a large variability of Saccharomyces strains (a total of 112, all of them identified as Saccharomyces cerevisiae) with no clear predominance of any strain throughout all the fermentation, probably related to the absence of killer phenotype and lack of previous inoculation with commercial strains.

DNA, Mitochondrial↗

PubMed2004

Microbiological characterization and probiotic potential of koko and koko sour water, African spontaneously fermented millet porridge and drink.

AIMS: To identify and examine the diversity of predominant lactic acid bacteria (LAB) in koko and koko sour water (KSW) from different Ghanaian production sites with regard to pattern of fermentation (API 50 CHL), genotype, antimicrobial activity, and resistance to low pH and bile salts. METHODS AND RESULTS: In total 215 LAB were isolated from koko and KSW. The isolates were identified using intergenic transcribed spacers (ITS)-PCR restriction fragment length polymorphism (RFLP), API 50 CHL, restriction enzyme analysis with pulsed-field gel electrophoresis (REA-PFGE) and sequencing of the 16S rRNA gene. The dominating micro-organisms in koko was found to be Weisella confusa and Lactobacillus fermentum, followed by Lact. salivarius and Pediococcus spp. Chemometric data analysis were used to link the LAB species to the different production stages and production sites. At intra-species level the isolates were found to have a great diversity. The isolates were investigated for antimicrobial activity using agar diffusion assays, and acid and bile tolerance. Most isolates showed low levels of antimicrobial activity towards the indicator strain Listeria innocua, but not towards the bacteriocin-sensitive Lact. sakei. Growth of all LAB isolates was unaffected by the presence of 0.3% (v/v) oxgall bile. The isolates were able to survive, but were not able to grow in growth medium adjusted to pH 2.5. CONCLUSIONS: The dominating LAB of koko and KSW were W. confusa and Lact. fermentum showing a pronounced taxonomic biodiversity at sub-species level between stages within the production as well as between production sites. Other species observed in KSW were Lact. salivarius, Ped. pentosaceus, Ped. acidilactici and Lact. paraplantarum. They occurred in levels of 108 CFU ml-1 in fresh KSW and showed uniform antimicrobial activity, and acid and bile tolerance. SIGNIFICANCE AND IMPACT OF THE STUDY: The present study gives a detailed picture of the taxonomy and diversity of LAB in an African-fermented millet product that may have potential as a probiotic product for the local population. The chemometric tools Principal Component Analysis and anova Partial Least Squares Regression were proven to be useful in the analysis of microbial groupings and associations with specific sites and stages in the production of koko and KSW.

Africa↗

PubMed2004

Genetic segregation of natural Saccharomyces cerevisiae strains derived from spontaneous fermentation of Aglianico wine.

AIMS: Investigation of the meiotic segregation of karyotypes and physiological traits in indigenous Saccharomyces strains isolated from Aglianico (South Italy) red wine. METHODS AND RESULTS: Segregation was studied in F1 and F2 descendants. Tetrads were isolated from sporulating cultures by micromanipulation. The spore clones were subjected to karyotype analysis by pulse-field gel electrophoresis (Bio-Rad model CHEF-DR II) and to various physiological tests. Certain chromosomes of the isolates showed 2:2 segregation patterns in F1 but proved to be stable in F2. The ability of cells to utilize maltose also segregated in a 2 : 2 manner in F1 and did not segregate in F2. Resistance to CuSO4, SO2 tolerance, the fermentative power and the production of certain metabolites segregated in both F1 and F2 generations and showed patterns indicating the involvement of polygenic regulation. CONCLUSIONS: The analysis revealed a high degree of genetic instability and demonstrated that meiosis can improve chromosomal and genetic stability. SIGNIFICANCE AND IMPACT OF THE STUDY: Winemaking is critically dependent on the physiological properties and genetic stability of the fermenting Saccharomyces yeasts. Selection of clones from F2 or later generations can be a method of reduction of genetic instability.

Chromosomes, Fungal↗

PubMed2002

Physiological diversity and trehalose accumulation in Schizosaccharomyces pombe strains isolated from spontaneous fermentations during the production of the artisanal Brazilian cachaça.

Twenty-seven Schizosaccharomyces pombe isolates from seven cachaça distilleries were tested for maximum temperature of growth and fermentation, osmotolerance, ethanol resistance, invertase production, and trehalose accumulation. Two isolates were selected for studies of trehalose accumulation under heat shock and ethanol stress. The S. pombe isolates were also characterized by RAPD-PCR. The isolates were able to grow and ferment at 41 degrees C, resisted concentrations of 10% ethanol, and grew on 50% glucose medium. Four isolates yielded invertase activity of more than 100 micromol of reducing sugar x mg(-1) x min(-1). The S. pombe isolates were able to accumulate trehalose during stationary phase. Two isolates, strains UFMG-A533 and UFMG-A1000, submitted to a 15 min heat shock, were able to accumulate high trehalose levels. Strain UFMG-A533 had a marked reduction in viability during heat shock, but strain UFMG-A1000 preserved a viability rate of almost 20% after 15 min at 48 degrees C. No clear correlation was observed between trehalose accumulation and cell survival during ethanol stress. Strain UFMG-A1000 had higher trehalose accumulation levels than strain UFMG-A533 under conditions of combined heat treatment and ethanol stress. Molecular analysis showed that some strains are maintained during the whole cachaça production period; using the RAPD-PCR profiles, it was possible to group the isolates according to their isolation sites.

Alcoholic Beverages↗

PubMed2000

The use of a starter culture in the fermentation of cassava for the production of "kivunde", a traditional Tanzanian food product.

Three cassava fermentation methods (spontaneous fermentation, back-slopping and the use of starter culture) for the production of kivunde, executed in three trials at 30 degrees C, were compared in terms of cyanide level reduction, microbiology and product quality improvement. Among the isolates from spontaneously fermented cassava batches, four strains were selected on the basis of their enzymatic activities and acid production. All were identified as Lactobacillus plantarum and were used as starters in this study. Lowest residual cyanide levels were detected after 120 h fermentation time in samples fermented with the starter culture and were below the maximum value of 10 mg/kg recommended by the Codex/FAO for cassava flour. This finding seems to be related to the alpha-glucosidase activity of the inoculated strains of which API-zyme (Bio-Merieux) tests showed activities of between 20 and > or = 40 nmol/4 h. The total residual cyanide levels of the spontaneous and back-slopping fermentations at 96 h were respectively 43.5 and 47.7 mg/kg dry weight of cassava. Extension of the fermentation period to 5 days, lead to further substantial reduction in the residual cyanide level in both these processes, but not below the recommended maximum value as in the case of starter culture fermented products. The spontaneous and back-slopping fermented cassava showed signs of deterioration after 3 days of fermentation. There was a sharp drop of pH and an increase of titratable acidity for all three batches during the first 48 h followed by a slow rise of pH and drop in titratable acidity towards the end of fermentation. The samples fermented with the starter culture had a smooth texture and pleasant fruity aroma, as opposed to the course and dull appearance and more complex flavour of the samples of spontaneous and back-slopping batches. During fermentation with starter culture, Enterobacteriaceae and yeasts and moulds could not be isolated throughout the period of fermentation (detection limit: 10 colony forming units/g). The present findings indicate the suitability of these Lb. plantarum strains as starter cultures for cassava fermentation in the kivunde process. The paper highlights the potential for the improvement of a traditional African fermented food (kivunde) through the use of a starter culture.

Bacteriological Techniques↗

PubMed2005

Analysis of yeast population during spontaneous alcoholic fermentation: effect of the age of the cellar and the practice of inoculation.

An ecological study of spontaneous fermentation in red wine carried out in two wineries located in the same village of the D.O.Ca. Rioja appellation (Spain). These two wineries were very different: one was very old and the other commenced its operations with the 2000 vintage. The study was conducted over seven consecutive years (1997-2003); the first 3 years in the old winery; and the other 4 years in the new one. Another difference between the wineries was the use of commercial strains: no starters were ever used in the old winery and two known strains (A and B) were employed only in a white must vat in the new winery. The number of different Saccharomyces cerevisiae strains detected for each vintage and the frequency of their appearance varied between the old winery and the new winery. There was no one clearly dominant clone in the fermentations studied in the old winery, whereas in the vinifications monitored in the new winery, a clear majority strain was detected. The same S. cerevisiae clone took over the spontaneous fermentation of all the vintages studied in this new winery. Analysis of the mtDNA restriction pattern of this strain revealed that it was identical to one of the commercial strain used (starter A), despite the fact that the fermentation studied was spontaneous. Microbial analysis conducted on winery equipment and winery surfaces prior to the 2003 vintage revealed that starter A had become a resident of the winery with a percentage of 50% among all S. cerevisiae. The presence of non-Saccharomyces yeasts in the early stages of vinification also differed between the old and new wineries.

Alcoholic Beverages↗

PubMed2006

Evolution of malolactic bacteria and biogenic amines during spontaneous malolactic fermentations in a Greek winery.

AIMS: To study the population dynamics of indigenous malolactic bacteria in a Greek winery and to examine their potential to produce detrimental levels of biogenic amines (BA) under winemaking conditions. METHODS AND RESULTS: Although the wines studied were of different vintage, grape variety and enological characteristics, molecular typing of malolactic bacteria revealed only a low number of strains within the single-species populations of Oenococcus oeni that developed during spontaneous fermentations. Strain MF1, originating primarily from the vineyards surrounding the winery invariably predominated in almost all samples. HPLC analysis showed a slight increase in the BA, putrescine, tyramine and phenylethylamine after malolactic conversion, while histamine, methylamine and ethylamine remained unaffected. No correlation could be established between the BA profiles and the bacterial compositions or the amino acid concentrations in wine samples studied. CONCLUSIONS: A certain regional bacterial flora is established in the winery that prevails in spontaneous malolactic fermentations (MLF) irrespective of the wine characteristics. In all cases, the BA content of the wines after malolactic conversion was within enologically acceptable levels. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report on the malolactic bacteria occurring naturally in spontaneous MLF in Greek red wines and a preliminary assessment of their impact on wine safety in relation to BA.

Biogenic Amines↗

PubMed2001

Genetic diversity of Saccharomyces cerevisiae strains during the 24 h fermentative cycle for the production of the artisanal Brazilian cachaça.

AIMS: Characterization of yeast populations and genetic polymorphism of Saccharomyces cerevisiae strains collected during the short fermentative cycles from the spontaneous fermentations during the artisanal cachaça production. METHODS AND RESULTS: The prevalent S. cerevisiae strains were analysed by PFG and RAPD-PCR using primers EI1 and M13. The molecular analysis have showed a high degree of genetic polymorphism among the strains within a 24 h fermentative cycle. CONCLUSION: The genetic diversity observed in the S. cerevisiae strains may be occurring due to the existence of a large number of individual genotypes within the species. The unique characteristics of the cachaça fermentation process probably allows for a faster detection of molecular polymorphisms of yeast strains than other types of fermentations. SIGNIFICANCE AND IMPACT OF THE STUDY: Spontaneous fermentations to produce cachaça, due to their characteristics, are an excellent model for the study of molecular diversity of S. cerevisiae strains during the production of fermented beverages.

Alcoholic Beverages↗

PubMed2004

Predominance of Saccharomyces uvarum during spontaneous alcoholic fermentation, for three consecutive years, in an Alsatian winery.

AIMS: The purpose of this study was to determine the origin of the yeasts involved in the spontaneous alcoholic fermentation of an Alsatian wine. METHODS AND RESULTS: During three successive years, must was collected at different stages of the winemaking process and fermented in the laboratory or in the cellar. Saccharomyces yeasts were sampled at the beginning and at the end of the fermentations. Saccharomyces cerevisiae clones were genetically characterized by inter-delta PCR. Non-S. cerevisiae clones were identified as Saccharomyces uvarum by PCR-RFLP on MET2 gene and characterized at the strain level by karyotyping. The composition of the Saccharomyces population in the vineyard, after crushing and in the vat was analyzed. This led to three main results. First, the vineyard Saccharomyces population was rather homogeneous. Second, new non-resident strains had appeared in the must during the winemaking process. Finally, the yeast population in the vat only consisted in S. uvarum strains. CONCLUSION: This 3-year study has enabled us to show the involvement of indigenous S. uvarum in the alcoholic fermentation. SIGNIFICANCE AND IMPACT OF THE STUDY: This study gives a first insight into the polymorphism of S. uvarum strains involved in a spontaneous alcoholic fermentation.

Chromosomes, Fungal↗

PubMed2006

Diversity of Saccharomyces strains on grapes and winery surfaces: analysis of their contribution to fermentative flora of Malbec wine from Mendoza (Argentina) during two consecutive years.

Spontaneous fermentations are still conducted by several wineries in different regions of Argentina as a common practice. Native Saccharomyces strains associated with winery equipment, grape and spontaneous fermentations of Malbec musts from "Zona Alta del Río Mendoza" region (Argentina) were investigated during 2001 and 2002 in the same cellar. Low occurrence of Saccharomyces on grapes and their limited participation during fermentation were confirmed. Strain sequential substitution during fermentation was observed. Between 30% and 60% of yeast population at the end of fermentation was coming from yeasts already present in the winery. A stable and resident Saccharomyces micro-flora in the winery was confirmed. It exhibited a dynamic behaviour during season and between years. Commercial strains were found during fermentation in different percentages, but their presence on winery equipment was low. The present work represents a first approach to winery yeast and spontaneous fermentation Saccharomyces population dynamics in an important viticultural region from Argentina that has never been characterized before. The results obtained have an important significance for the local industry, showing for the first time the real situation of the microbial ecology of alcoholic fermentation in an industrial winery from Mendoza, Argentina.

Argentina↗

PubMed1993

Occurrence of killer yeasts in spontaneous wine fermentations from the tuscany region of Italy.

The occurrence of killer yeasts in an area of Tuscany (central Italy) was studied. Killer yeasts were found in 88% of spontaneous wine fermentations from 18 wineries. The incidence of killers varied with respect to fermentation stage and vintage period, increasing from the first vintage to successive ones and from the commencement to the end of fermentation. At the end of fermentation, the proportion of killer strains relative to total yeast population was below 25% in 15 cases, above 75% in 6 cases, from 25 to 50% in 5 cases, and from 50 to 75% in 3 cases. Karyotype analysis also showed a mixed killer population in the fermentations in which the killers dominated.

Journal Article↗

PubMed2003

Volatile compounds produced by Lactobacillus fermentum, Saccharomyces cerevisiae and Candida krusei in single starter culture fermentations of Ghanaian maize dough.

AIMS: To identify and compare the volatile compounds associated with maize dough samples prepared by spontaneous fermentation and by the use of added starter cultures in Ghana. METHODS AND RESULTS: The starter cultures examined were Lactobacillus fermentum, Saccharomyces cerevisiae and Candida krusei. For identification of aroma volatiles, extracts by the Likens-Nickerson simultaneous distillation and extraction technique were analysed by gas chromatography-mass spectrometry (GC-MS) and using a trained panel of four judges by GC-Olfactometry (GC-sniffing). Compounds identified by GC-MS in maize dough samples after 72 h of fermentation included 20 alcohols, 22 carbonyls, 11 esters, seven acids, a furan and three phenolic compounds. Of the total 64 volatile compounds, 51 were detected by GC-sniffing as contributing to the aroma of the different fermented dough samples. Spontaneously fermented maize dough was characterized by higher levels of carbonyl compounds while fermentations with added L. fermentum recorded the highest concentration of acetic acid. S. cerevisiae produced higher amounts of fusel alcohols and increasing levels of esters with fermentation time and C. krusei showed similarity to L. fermentum with lower levels of most volatiles identified. CONCLUSION: The present study has given a detailed picture of the aroma compounds in fermented maize and demonstrated that the predominant micro-organisms in fermented maize dough can be used as starter cultures to modify the aroma of fermented maize dough. SIGNIFICANCE AND IMPACT OF THE STUDY: The study has documented the advantage of using starter cultures in African traditional food processing and provided a scientific background for introducing better controlled fermentations.

Alcohols↗