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• v.11(12); 2010 Dec

Fermenting knowledge: the history of winemaking, science and yeast research

Paul j chambers.

1 Australian Wine Research Institute, PO Box 197, Glen Osmond, Adelaide, SA 5064, Australia

Isak S Pretorius

In the second article of the ‘Food and Science' series, Paul Chambers and Isak Pretorius explain the central role of yeast in wine making and how biotechnology can contribute to improving the quality of wine.

Wine has been with us since the dawn of civilization and has followed humans and agriculture along diverse migration paths ( Fig 1 ). Serendipity presumably played a part in its genesis more than 7,000 years ago: damaged grapes spontaneously fermented in harvesting vessels; curious farmers tasted the resultant alcoholic beverage; the curious farmers liked what they tasted and enjoyed its effects; said farmers preferred fermented grape juice to the unfermented fruit. The fate of the grape was sealed.

One might argue that the most important test tube in the birth and growth of the modern life sciences is the fermenter…

A generalized scheme of the spread of Vitis vinifera noble varieties of grapevine and winemaking from their centre of origin in Asia Minor to other parts of the world.

One might argue that the seeds of science and technology, particularly biotechnology, were also sown at this time. Empirical observations of natural events and processes were harnessed in repeat ‘experiments'—which is to say, vintages—and improvements were made by trialling modifications to practices, retaining those that were beneficial and discarding failures, with the results communicated down through the generations. At that time, there was no EMBO reports or alternative means by which to facilitate horizontal dissemination of information, but the principle of development— sans peer review—is clear: experimentation and invention lead to progress—technological and otherwise—and new knowledge is shared and built upon.

Of course, early inventions and innovations in grape and wine production were based on little or no knowledge of the biology of grapevines or the microbes that drive fermentation. In fact, it would be several thousand years before it was even known that microscopic organisms exist: using a primitive microscope, Antonie van Leeuwenhoek observed cells for the first time in 1680 ( Fig 2 ).

Selected milestones that mark the path of research in microbiology and yeast biology that have affected, directly or indirectly, wine science and winemaking.

Scientific knowledge grows at an exponential rate, and nowhere is this more evident than in the historical milestones of chemistry and biology that have shaped our understanding of the biology of the microorganisms that drive fermentation ( Fig 2 ). This progress has been adorned with some of the most significant names in the chemical and biological sciences, including van Leeuwenhoek, Lavoisier, Gay-Lussac, Pasteur, Buchner and Koch. One might argue that the most important test tube in the birth and growth of the modern life sciences is the fermenter, and the most important model organism has been the yeast Saccharomyces cerevisiae —commonly known as baking, brewing or wine yeast. As readers might know, this is exemplified in the origin of the word enzyme—‘en' meaning within and ‘zyme' meaning leaven. Yeast has been integral to pioneering work in microbiology and biochemistry, particularly in the fields of metabolism and enzymology ( Barnett, 1998 , 2000 ; Barnett & Lichtenthaler, 2001 ).

Throughout the early decades of the twentienth century the place for S. cerevisiae in fundamental research was affirmed, and there are several good reasons for this. Our close relationship with this yeast in food and beverage production over millennia tells us that it is safe to work with; as confirmed by its ‘Generally Recognised as Safe' designation by the US Food and Drug Administration. In addition, it is inexpensive, easy to grow and can be stored for long periods in suspended animation. Perhaps the most important thing is that it has accessible genetics that can be followed through sexual and asexual cycles ( Barnett, 2007 ).

The 1970s set the stage for another explosion of knowledge, sparked by the advent of gene technology and driven by a convergence of genetics, biochemistry, cell biology, microbiology, physical and analytical chemistry, as well as computing brought together under the banner of molecular biology ( Fig 3 ). Yeast molecular biology was established when Gerald Fink's group in the USA demonstrated that yeast could be transformed with foreign DNA ( Hinnen et al, 1978 ). In the same year, Jean Beggs in the UK developed a shuttle vector between Escherichia coli and S. cerevisiae that enabled cloning in yeast ( Beggs, 1978 ). The research community now had a eukaryotic host that was amenable to genetic engineering, benefiting both fundamental research and offering the potential of precise engineering of novel strains for industrial applications. It was the first host cell for industrial-scale production of a recombinant vaccine against hepatitis B and a recombinant food-grade enzyme, chymosin, which is used in cheese processing ( Pretorius et al, 2003 ).

Selected milestones that mark the path of research in genetics and molecular biology that have affected, directly or indirectly, wine science and winemaking.

Ever since, S. cerevisiae has been one of the most important model organisms in molecular biology and emerging fields; breakthroughs and technological advances in molecular, systems, and now synthetic biology rarely happen without S. cerevisiae figuring somewhere prominently in the story ( Fig 3 ). The international yeast science community has been particularly progressive and proactive in establishing large collaborative projects and building resources that are available to the scientific community. S. cerevisiae was the first eukaryote to have its genome sequenced ( Goffeau et al, 1996 ), a feat that was achieved through an international effort that involved 600 scientists, which paved the way for the first chip-based gene array experiments ( Schena et al, 1995 ). It was the first organism to be used to build a systematic collection of bar-coded gene deletion mutants ( Winzeler et al, 1999 ; Giaever et al, 2002 ), in which there are deletion strains for most of the open-reading frames in the S. cerevisiae genome. This has enabled high-throughput functional-genomic experiments, and anyone seeking information on just about any aspect of S. cerevisiae biology has access to the amazing community resource: the Saccharomyces Genome Database (SGD; http://www.yeastgenome.org/).

All of this is important to wine research; our winemaking workhorse is centre stage in thousands of research projects worldwide, so we know more about this humble eukaryote than any other organism on the planet. It is therefore unsurprising that wine research has benefited enormously from the privileged place that S. cerevisiae occupies in life sciences research. This is particularly evident in the impact that advances in molecular biology and related fields have had on winemaking.

In the hands of molecular biologists, S. cerevisiae is the most tractable of organisms; it is amenable to almost any modification that modern biology can throw at a cell. This makes it an ideal host for generating variants with improved and even exotic phenotypes that will benefit winemaking. The following gives some examples of current research and directions in this field.

In modern winemaking, fermentations are driven largely by single-strain inoculations; pure cultures of selected strains of S. cerevisiae are added to grape must as soon as possible after crushing. This ensures greater control of vinification, leads to more predictable outcomes and decreases the risk of spoilage by other microorganisms. There are many—probably hundreds of—different yeast strains available, and the winemaker's choice can substantially effect the quality of the wine ( Lambrechts & Pretorius, 2000 ; Swiegers et al, 2005 ).

One of the reasons for the yeast-induced variation in wine quality is that, during fermentation, S. cerevisiae produces an abundance of aroma-active secondary metabolites and releases many aroma compounds from inactive precursors in grape juice, which greatly affect the sensory properties of the wine ( Swiegers & Pretorius, 2007 ). Thus, any genetic variation in wine yeast that affects the production or release of sensorially important molecules will affect wine quality. In this context it has been demonstrated, for example, that different commercial yeast strains generate wines with very different profiles of volatile thiols ( Swiegers et al, 2009 ). These thiols—which are present in grape juice as non-volatile cysteinylated precursors ( Tominaga et al, 1998 )—are often described as ‘passionfruit', ‘tropical fruits' and ‘citrus' by tasters, flavours that are particularly important in wine varieties such as Sauvignon Blanc ( Dubourdieu et al, 2006 ).

Molecular biology and its tools are crucial to our understanding of the genetic and molecular bases of yeast-driven volatile thiol release from non-volatile precursors in grape juice. Howell et al (2005) have used bioinformatic tools and the SGD to identify candidate S. cerevisiae carbon–sulphur lyase genes that might be involved in the release of volatile thiols from cysteinylated precursors during fermentation. The researchers used targeted gene deletion to remove these candidate carbon–sulphur lyases from the wine and laboratory yeast strains, and they identified four genes that potentially contribute to the release of these important aroma molecules.

Swiegers et al (2007) then engineered a wine yeast, VIN13, to constitutively express a carbon–sulphur lyase gene, tna A, from E. coli . Sensory analysis revealed that, compared with its non-engineered relative, this transgenic yeast, VIN13 (CSL1), had a positive impact on the release of volatile thiols from a Sauvignon Blanc grape juice. The authors commented that wine assessors preferred the VIN13 (CSL1)-derived experimental wines to the relatively neutral VIN13-derived wines.

A similar approach has been used to engineer yeasts for the enhanced production of fruity esters ( Lilly et al, 2006a ) and to increase the production of higher, fusel alcohols ( Lilly et al, 2006b )—all of which contribute to the flavour profiles of wines. Although this work is in the early stages of development, it shows the value of yeast molecular biology, and the amazing resources that come with it.

Wine alcohol content is of growing importance to the wine industry. In some wine regions, it has been increasing during recent decades ( Godden & Muhlack, 2010 ). The main reason for this increase is that grapegrowers tend to leave fruit on the vine as long as possible to increase fruity characters—which develop as berries mature—and reduce undesirable ‘green' characters. This practice, however, produces fruit with a higher sugar content, which translates to higher ethanol concentrations in the wine.

A recent review by Kutyna et al (2010) discusses several metabolic engineering strategies that have been explored to generate wine yeasts that can divert some carbon metabolism away from ethanol production, with the aim of decreasing ethanol yields during vinification. Understanding the central metabolism of yeast and the genes that drive it has been crucial to this work. Candidate genes that are likely to influence ethanol yields can be identified from a range of sources, including the SGD, and then manipulated and cloned as required. Several laboratories have targeted the glycerol-3-phosphate dehydrogenase isozymes GPD1 and GPD2 , which divert carbon from glycolysis to glycerol production ( Michnick et al, 1997 ; Remize et al, 1999 ; de Barros Lopes et al, 2000 ).

Increased expression of either of the GPD paralogues increased glycerol and decreased ethanol yields. However, increased Gpd activity also led to increased amounts of acetic acid in the fermentation product. This was probably owing to rectification—by one or more of the five aldehyde dehydrogenase isozymes—of a redox imbalance that resulted from excessive Gpd-driven oxidation of NADH. Aldehyde dehydrogenase isozymes drive the oxidation of acetaldehyde to acetic acid with concomitant reduction of coenzymes NAD + or NADP, depending on which isozyme is involved ( Navarro-Aviño et al, 1999 ). This might be good for a yeast cell struggling with an imposed redox imbalance, but an increase in acetic acid production is not good news for winemakers; excessive vinegar is not desirable in wine. This problem was alleviated by knocking out one of the five aldehyde dehydrogenase isozymes, ALD6 ( Eglinton et al, 2002 ; Cambon et al, 2006 ).

Similar approaches have targeted S. cerevisiae pyruvate decarboxylase isozymes, alcohol dehydrogenase isozymes and glycerol transporters, leading to increased glycerol yields and reduced ethanol production ( Kutyna et al, 2010 ). However, while there are probably several good candidate ‘low-ethanol' wine yeast strains sitting in various labs around the world, none have been tested in commercial-scale, industrial fermentations. This is largely because consumers are generally unaccepting of genetically modified organisms (GMOs) in foods and beverages.

Another area of ongoing research in wine yeast molecular biology is the development of strains that flocculate—that is, form clumps—at the end of fermentation. This facilitates the process of settling them out of suspension and separating them from the wine, thereby reducing the need for clarification. The timing of flocculation is crucial; it must not happen too early, as yeast in large flocs are inefficient at sugar utilization and can generate suboptimal—stuck or sluggish—fermentations ( Pretorius, 2000 ).

Generally, wine yeasts are not good at flocculation; they do not form large clumps that settle out of suspension. Many years of research using laboratory strains of S. cerevisiae led to the identification and characterization of genes that encode cell-surface glycoproteins—including lectin-like flocculins—that cause, among other things, flocculation and subsequent settling to the bottom of the fermentation vessel ( Pretorius, 2000 ).

Recent findings have identified a problem with extrapolating basic research on laboratory strains to those used in industry; yeasts domesticated for different purposes have different phenotypes. Work by Govender et al (2008) on the flocculation genes FLO1 , FLO5 and FLO11 , for example, demonstrated the potential ability of engineered ADH2 - or HSP30 -promoter/ FLO gene combinations to switch on flocculation at the end of fermentation; ADH2 and HSP30 are both upregulated in stationary-phase cells, so their promoters are suitable candidates to drive the expression of genes in later stages of wine fermentation.

The results of this work were promising, but, when they were carried over to wine yeast, the findings were rather different. There were even substantial differences between wine yeast strains, leading the authors to caution that “optimisation of the flocculation pattern of individual commercial strains will have to be based on a strain-by-strain approach” ( Govender et al, 2010 ). Nonetheless, controlled expression of FLO genes at the end of fermentation remains a plausible technique for improving the performance of wine yeast, but the strategies required to achieve a desirable outcome might be more complex than was originally thought.

While the complexity of biological systems is a cause for excitement and wonder to most biologists, it can make engineering novel strains for industrial applications trickier than molecular biology and biotechnology textbooks might suggest. For those of us working on industrial yeast strains, it might be pertinent to directly tackle the issue of complexity and use systems biology approaches to better understand the workings of yeast metabolism. This should lead to more accurate modelling of metabolic processes for better-informed manipulations, to achieve targeted, predictable outcomes.

However, molecular biologists face one important obstacle to this progress: near worldwide refusal to permit the use of GMOs in the production of foods and beverages…

S. cerevisiae has been at the forefront of ‘-omics' research. This provides us with enormous opportunities to improve understanding of wine yeast complexity, which, in turn, will inform the design of new strains for industrial applications. Increased and improved knowledge from a huge number of studies investigating strains of S. cerevisiae at the various -omic levels gives wine yeast scientists a head start in this field ( Borneman et al, 2007 ; Petranovic & Vemuri, 2009 ).

One of the most interesting developments has come from the sequencing of a wine yeast genome, and its comparison with the genomes of a laboratory strain and an opportunistic pathogenic S. cerevisiae ( Borneman et al, 2008 ). The authors found a difference of about 0.6% in sequence information between the wine yeast and the other strains. They also found, perhaps more importantly, 100 kb of additional genome sequence in the former; enough to carry at least 27 genes. Open reading frames (ORFs) in the additional sequences do not resemble anything found in other strains of S. cerevisiae , but seem to be similar to genes found in distant fungal relatives. BLAST searches have indicated that some of the genes that are specific to wine yeast are similar to those encoding cell-wall proteins. This might contribute to the greater robustness of wine yeast, compared with laboratory strains. Other genes might encode proteins associated with amino acid uptake, which is significant in the context of wine sensory attributes; amino acid metabolism is central to the production of many sensorially important volatile aroma compounds.

Novo et al (2009) published similar findings from a different wine yeast strain (EC1118) and suggested that the extra sequence was probably the result of horiziontal gene transfer. Further work using functional genetics—to determine the effects of knocking out and overexpressing the ORFs—should enable characterization of the phenotypes of these ORFs, determine their relevance in the context of winemaking and might also reveal their origins.

There have also been numerous studies describing transcriptomic, proteomic and metabolomic analyses of wine-yeast fermentations. This work is beginning to provide insights into wine-yeast fermentations, but it is still early days. It should also be noted that much of the -omics work on wine yeast has used resources and databases that are based on laboratory strains. It is now clear that there are genomic differences between wine and lab strains of S. cerevisiae , and these might affect -omics data acquisition and analysis. For example, gene-array chips based on the reference laboratory strain S288c will not include the additional ORFs found in wine strains. This does not suggest that earlier work is invalid, but that there are likely to be gaps in it.

As the various -omics fields progress, it should be possible to build systems-based mathematical models of metabolism that will facilitate the in silico design of new wine yeast strains ( Borneman et al, 2007 ). In parallel, we see the emergence of synthetic biology where, yet again, S. cerevisiae is a key player. It should not be too long before we have customised S. cerevisiae genomic components—regulatory elements to control the expression of targeted genes, or cassettes carrying genes encoding metabolic pathways to shape wine-relevant traits, for example—available ‘off the shelf' for designing, building and refining metabolic processes in our wine yeast. But are consumers ready for this brave and exciting new world?

The engineered wine yeast strains described in this paper show the potential of novel yeast strain development to improve wine quality. But molecular biologists face a major obstacle to this progress: near world-wide refusal to permit the use of GMOs in the production of foods and beverages, at least in ‘developed' countries ( Gross, 2009 ; Pretorius & Høj, 2005 ). Wine industries in most parts of the world have eschewed the use of GMOs in commercial winemaking, leaving most new-generation wine yeasts on the laboratory shelf, where they await more enlightened times.

Two genetically modified wine yeast strains have been released to market in a limited number of countries including the USA, Canada and Moldova: ML01 and 522 EC− . ML01, a transgenic wine yeast, has genes that enable it to perform malolactic fermentation (MLF), a deacidifying secondary fermentation in which malic acid—present in grape juice—is decarboxylated to lactic acid. MLF is usually performed by the lactic acid bacterium Oenococcus oeni after alcoholic fermentation. However, this bacterium is rather fastidious, being inhibited by a range of conditions that are typical of fermented grape juice—low pH, high alcohol content, poor nutrient availability and the presence of sulphur dioxide—and can become ‘stuck' or take considerable time to complete fermentation ( Davis et al, 1985 ). In addition, lacitic acid bacteria can produce a range of biogenic amines, which are associated with health risks ( Lonvaud-Funel, 2001 ).

A wine yeast that completes both primary and secondary fermentations should therefore have great potential in the wine industry. The genetically modified wine yeast ML01 carries two foreign genes—the Schizosaccharomyces pombe malate transporter gene ( mae1 ) and the O. oeni malolactic enzyme gene ( mleA )—which are both chromosomally integrated and regulated by the S. cerevisiae PGK1 promoter and terminator ( Husnik et al, 2006 ). This enables the host wine yeast to perform MLF, in parallel with alcoholic fermentation.

The researchers went to great lengths to ensure the safety of ML01. The transgenes came from microorganisms found in wine, there were no antibiotic resistance genes or vector sequences carried by the yeast and transcriptome and proteome analysis showed no important differences in gene expression profiles between the genetically modified strain and its parent. The FDA granted ‘Generally Regarded As Safe' status to ML01, but it has not been widely adopted, even in countries where it is approved for use. This is largely owing to concerns about export markets that do not tolerate GMOs. In fact, wine industries in many countries have banned the use of GMOs in wine production, in order to avoid jeopardizing their exports. ​ exports.

The genetically modified wine yeast 522 EC− was engineered to reduce the risk of ethyl carbamate production during fermentation. Ethyl carbamate, a potential carcinogen, is the product of yeast-derived urea reacting with ethanol. It is usually produced at such low levels—if at all—that it is not a cause for concern, but it sometimes can make an appearance in some wine-producing regions.

S. cerevisiae is able to degrade urea before it is secreted and release ammonia instead, thereby reducing the risk of generating ethyl carbamate. This is achieved by the action of an enzyme encoded by DUR1,2 , but this gene is repressed by nitrogen and therefore downregulated throughout much of wine fermentation. Coulon et al (2006) placed a copy of DUR1,2 behind a constitutive ( PGK1 ) S. cerevisiae promoter, which led to a reduction in ethyl carbamate yields. Interestingly, this genetically modified yeast is self or cis cloned; it carries no foreign DNA and therefore is not transgenic. Nonetheless, because it was generated by using techniques that involved the manipulation of DNA in vitro , the regulations of many countries classify it as a GMO. Again, to the best of our knowledge, this yeast is not being used in the industry. This might be because ethyl carbamate production is not a widespread problem, but it probably also reflects the influence of GMO bans and the reluctance of winemakers to risk losing market share in countries that harbour strong anti-GMO sentiment.

Who knows what bottled masterpieces await us as we sculpt novel yeast strains in the laboratory using molecular, systems and synthetic biology

Winemaking, science and technology have interwoven histories and have grown together over the millennia, benefiting from each other. Although science is an important part of an oenologist's training and scientific methods and equipment are routinely employed in the winery, winemakers are not scientists per se . They are, perhaps more appropriately regarded as artisans, with the emphasis on the ‘art'. As for many human endeavours, the Arts progress with developments in technology; think of the use of acrylic paint in the fine arts since its introduction in the 1950s, or David Hockney's use of a Polaroid camera to create photocollages. In the way that acrylic paint and photography have provided more options to artists, enabling them to broaden their horizons, yeast science and technology is adding to the winemaker's palette. Who knows what bottled masterpieces await us as we sculpt novel yeast strains in the laboratory using molecular, systems and synthetic biology. The only real obstacle that we face is consumer acceptance of GMOs; we can only hope that rationality will eventually prevail.

Science & Society Series on Food and Science

This article is part of the EMBO reports Science & Society series on ‘food and science' to highlight the role of natural and social sciences in understanding our relationship with food. We hope that the series serves a delightful menu of interesting articles for our readers.

Acknowledgments

Research at the Australian Wine Research Institute (AWRI) is financially supported by Australia's grapegrowers and winemakers through their investment body, the Grape and Wine Research Corporation, with matching funding from the Australian Government. Systems biology research at the AWRI uses resources provided as part of the National Collaborative Research Infrastructure Strategy (NCRIS), an initiative of the Australian Government, in addition to funds from the South Australian State Government. AWRI's collaborating partners within this NCRIS-funded initiative—which is overseen by Bioplatforms Australia—are Genomics Australia, Proteomics Australia, Metabolomics Australia (of which the Microbial Metabolomics unit is housed at the AWRI) and Bioinformatics Australia.The AWRI is part of the Wine Innovation Cluster in Adelaide.

The authors declare that they have no conflict of interest.

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The article presents review on potential of wine production from Banana. Traditional alcoholic beverage has become common because of economic issue. This work was aimed to improve production process of alcoholic beverage based banana extract and to evaluate sensory parameters of the obtained alcoholic beverage. Juice was extracted from banana (Musa sapientum) pulp with was inoculated with Baker's yeast (Saccharomyces Cerevisiae)and naturally grown mycelium and held at 30+2'c for 14 days. Banana, a wonderfully sweet fruit with firm and creamy flesh that come prepackaged in a yellow jacket, available for harvest throughout the year consists mainly of sugars and fibers which make it a source of immediate and slightly prolonged energy. When consumed, reduces depression, anemia, blood pressure, stroke risk, heartburns, ulcers, stress, constipation and diarrhea. The physical chemical parameters was determine during the fermentation using a standard procedures like pH , type of alc...

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Banana, a wonderfully sweet fruit with firm and creamy flesh that come prepackaged in a yellow jacket, available for harvest throughout the year consists mainly of sugars and fibers which make it a source of immediate and slightly prolonged energy. When consumed, reduces depression, anemia, blood pressure, stroke risk, heartburns, ulcers, stress, constipation and diarrhea. It confers protection for eyesight, healthy bones, kidney malfunctions, morning sickness, itching and swelling, improves nerve functions as well as help people trying to give up smoking. Fermentation of banana must for 144 h was carried out using recipes A to D. Recipe A contained a mixture of banana must with natural yeast. A was enhanced with granulated sugar to obtain recipe B. Recipe C contained recipe A augmented with granulated sugar and bakers' yeast while recipe D (control) contained only granulated sugar solution and bakers' yeast. Wine produced had values that ranged from 31.4 ± 0.29 to 33.2 ± 0.12°C for temperature, 3.38 ± 0.017 to 3.54 ± 0.052 for pH, 0.999 ± 0.0085 to 1.02 ± 0.0058 for specific gravity, 0.586 ± 0.018 to 0.71 ± 0.017 for optical gravity, 1.37 ± 0.075 to 1.383 ± 0.152 for percentage (%) alcohol (v/v), 0.271 versus 0.012 to 1.348 ± 0.072 for percentage (%) titratable acidity, 8.2 ± 0.099 to 9.38 ± 0.283 for total aerobic counts and 3.5 ± 0.5 to 4.75 ± 0.1 for R f. Malo-lactic fermentation after 48 h was evident. Taste testing showed very little differences in wines from recipes A to C. Statistical analyses of tested parameters at 95% confidence level showed no significant differences. The wine from the control was similar to natural palm wine in taste and characteristics. Wine could thus be produced from banana for immediate consumption, within 48 h, using the recipes A to C.

Srinivas Tadepalli

Juice was extracted from banana (Musa sapientum) pulp with the addition of lemon juice and was inoculated with Baker‟s yeast (Saccharomyces cerevisiae) and held at 30±2c for seven days. The result of the yeast count increases at 48hr, and at 96hr the yeast count decreased gradually. It ranges from 4.9x10 7 cfu/ml at 0hr, 5.1x10 7 at the 48hr and 4.8x10 7 cfu/ml at 168hr. The pH of the Banana wine produced at the end of fermentation decreased (2.85) while the testable acidity of the Banana wine produced increased. The total dissolved solids, total suspended solids decreased with increasing length of the fermentation time of juice. The alcohol content of the wine increased with 14%.

Constance C Ezemba

This research was carried out to produce wine from banana (Musa sapientum) pulp using yeast (Saccharomyces cerevisiae) isolated from grape (Vitis vinifera).The fermentation of the banana wine lasted for 21 days. Physico-chemical parameters were determined during fermentation using standard procedures. Liquor of the fermenting must was removed for every 48 hours from the fermentor for analysis of pH, titratable acidity, specific gravity and reducing sugar using standard procedures. The results from the experiment showed that specific gravity of the wine was observed to reduce drastically as the fermentation progressed. The pH of the banana wine during fermentation increased from 4.16-4.22 at day to the last day while the titrable acidity (% w/v tartaric acid) of the banana wine produced increased from 1.05-1.77. The alcohol content of the wine increased with time. The specific gravity values were observed to range from 1.266 to 1.184 kg/m 3 which gradually decreased throughout the period of fermentation. At the end of the fermentation,

Asian Journal of Dairy and Food Research

Background: Banana is a commercial fruit of many tropical and sub-tropical nations. India is the largest producer of banana in the world. The fruit is a valuable source of carbohydrates, phenolics compounds, minerals and dietary fibre. Due to bumper production, seasonality and market gluts, huge post-harvest losses are recurring phenomena in many banana growing states in India. Hence, the present study was undertaken to evaluate the suitability of some commercial banana cultivars in the development of wine and to evaluate the qualitative changes during ageing. Methods: Wine was prepared from banana varieties (Poovan, Grand Naine, Yangambi (KM-5), Karpooravalli and Palyankodan) in 1:1 and 1:2 dilutions of pulp and water. The TSS content of the substrate was raised to 20°Brix, followed by addition of yeast @ 1.25 g/l. The must was treated with Potassium Metabisulphite (KMS) @ 0.05 g/L to inhibit the growth of other microorganisms. Fermentation was carried out for 15 days, followed by ...

NSHIMIYIMANA Pacifique

The research was conducted on the production of banana liqueur form AAA-EAH beer bananas [Intuntu]. The study started by the incubation of mature green banana at 200C for 4 days of ripening. After the ripen banana fruits were crushed with spear glass (Impeta cylindica) to obtain juice which was pasteurized at 80oC for 15 min. The inoculation was made with Saccharomyces cerevisiae then incubated at 25oC, after 48 hours the fermentation was done with no more production of CO2. The wine produced was 9% (v/v) of alcohol content and then proceeded to distillation at 78-82oC for the production of a brandy which was 89.5% (v/v) of alcoholic strength after check. The brandy was again macerated in banana juice with 150 g of Gros Michel banana fruits and 10 g of honey for sweetening and aromatization. The maceration was kept first at temperature below 12oC for 24 hours and the second 24 hours at 25oC for sweetness and aromas extraction, the produced liqueur was then filtrated and bottled ready for analysis. The 30.19oBrix produced liqueur was checked for physicochemical properties and proved to have 40.5% (v/v) of alcoholic strength with the total acidity of 2.5 g/l, a volatile acidity of 1.03 g/l and a density of 1.045 which made the study to be classed as successful. Keywords: AAA-EAH beer bananas/Musa spp, alcohol strength, Saccharomyces cerevisiae, brandy

Karan Bagde , pranali waghdhare

Banana plant is one of the largest herbaceous flowering plant which serves as an ideal and low-cost food source for third world and developing countries. Every part of the Banana plant can be used as Functional food. Apart from the fruit that is the edible part, there are various organic compounds in the banana peel. Although these compounds has many health-beneficial components it is treated as waste , thus arises an issue for the disposal of these peels. The powder is rich in micronutrients which can be utilized to produce value-added products there by the banana peel can act as a functional ingredient. This would lead to waste utilization, thereby addressing the issue of waste disposal. The study aims to develop a banana wine with honey which is incorporated with banana peel powder it and evaluate the physicochemical, microbial characteristics of the products. The variants of banana wine enriched with the banana peel and honey, used as the sugar substitute. The banana juice with 50% of honey and three concentrations (2%, 4%, 6%) of banana peel powder inoculated with 2% of Baker’s yeast i.e. (Saccharomyces cerevisiae) and held at 28±20C. Temperature maintained at 28±20C for the fermentation period. Soluble solids, pH, and specific gravity were all monitored for the fermentation period. The wine produce with 5% alcohol, 1% titrable acidity, and also the micronutrients like Ca, K, Fe, P recorded for the 3 days’ time interval. The number of micronutrients in the wine increased with the increase in fermentation.

Saikat Mazumder

SpringerPlus

Ositadinma Ugbogu , CHUKWUMA EZEONU , Alloysius Chibuike Ogodo

Pawpaw, banana and watermelon are tropical fruits with short shelf-lives under the prevailing temperatures and humid conditions in tropical countries like Nigeria. Production of wine from these fruits could help reduce the level of post-harvest loss and increase variety of wines. Pawpaw, banana and watermelon were used to produce mixed fruit wines using Saccharomyces cerevisiae isolated from palm wine. Exactly 609 and 406 g each of the fruits in two-mixed and three-mixed fruit fermentation respectively were crushed using laboratory blender, mixed with distilled water (1:1 w/v), and heated for 30 min with subsequent addition of sugar (0.656 kg). The fruit musts were subjected to primary (aerobic) and secondary (anaerobic) fermentation for 4 and 21 days respectively. During fermentation, aliquots were removed from the fermentation tank for analysis. During primary fermentation, consistent increases in alcohol contents (ranging from 0.0 to 15.0 %) and total acidities (ranging from 0.20 to 0.80 %) were observed with gradual decrease in specific gravities (ranging from 1.060 to 0.9800) and pH (ranging from 4.80 to 2.90). Temperature ranged from 27 °C to 29 °C. The alcoholic content of the final wines were 17.50 ± 0.02 % (pawpaw and watermelon), 16.00 ± 0.02 % (pawpaw and banana), 18.50 ± 0.02 % (banana and watermelon wine) and 18.00 ± 0.02 % (pawpaw, banana and watermelon). The alcoholic content of the wines did not differ significantly (p > 0.05). The pH of all the wines were acidic and ranged from 2.5 ± 0.01 to 3.8 ± 0.01 (p > 0.05). The acid concentration (residual and volatile acidity) were within the acceptable limit and ranged from 0.35 ± 0.02 to 0.88 ± 0.01 % (p > 0.05). Sensory evaluation (P > 0.05) rated the wines acceptability as 'pawpaw and banana wine' > 'pawpaw and watermelon' > 'pawpaw, watermelon and banana' > 'banana and watermelon wine'. This study has shown that acceptable mixed fruit wines could be produced from the fruits with S. cerevisiae from palm wine.

The Pharma Innovation Journal

Manjusha Nevase

Asian Journal of Applied Science and Technology (AJAST)

AJAST Journal

Wine is a fermented drink made by the controlled culture of yeasts on fruit juices. This study was undertaken to produce acceptable wines from blends of banana and pineapple by the fermentative action of Meyerozyma guilliermondii strain 1621 and Pichia guilliermondii strain PAX-PAT 18S. The fermentation process lasted for a period of 28 days and, the aging process was for 2 months. The fermentation process comprised two set ups- one was fermented by Meyerozyma guilliermondii strain 1621 and the other was fermented by Pichia guilliermondii strain PAX-PAT 18S. The process was monitored and controlled by carrying out physicochemical analysis (pH, temperature, specific gravity, total titratable acidity, and alcohol content) and yeast count using standard methods. There was a decrease in the pH for both wines and an increase in the total titratable acidity. The temperature was between 17 and 27 0C for both wines. The specific gravity of the wines decreased during the fermentation leading to an increase in alcohol production. There was an increase in yeast count from 6.7×107 sfu/ml to 1.8×108 sfu/ml between days 1 and 17 and a decrease from 1.8×108 sfu/ml to 0 sfu/ml between days 17 to 85 for Meyerozyma guilliermondii; also an increase from 5.1×107 sfu/ml to 1.7×108 sfu/ml from day 1 to 17, and a decrease from 1.7×108 sfu/ml to 0 sfu/ml between day 17 to 85 for Pichia guilliermondii. Statistically, there was no significant difference between the yeast counts, temperature, pH, total titratable acidity, and specific gravity but there was signa ificant difference between the alcohol production for both wines. This study shows that wines can be successfully produced using Meyerozyma guilliermondii strain 1621 and Pichia guilliermondii strain PAX-PAT 18S.

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