AN INVESTIGATION OF FERMENTATION RATE ON SACCHAROMYCES CEREVISAE AND SACCHAROMYCES SP.

Nguyen Thi Anh Thoa, Nguyen Ba Ngoc

Abstract: Red wine is a light wine which was fermentated by fruit juice with high level of nutrient, nice taste, and have good effect for the health when we use it with a right amount. In the World, specially in Europe, wine has been made a long time ago, and it was used very commonly. But in Vietnam, wine products have not been concerned much, pruducing scale is small and artisanal; its efficiency is low. During the traditional wine making, wine is fermentated by natural types of yeasts which stay in the grapes. To have a good quality of product, researches should be done to categorize and choose the best yeast genera. Until now, there are lots of experiments in wine making process from Saccharomyces.cerevisiae, which was sold in the market to make bread. So this thesis chose Saccharomyces.cerevisiae and Saccharomyces spp (yeast on the husky of grape) to do the reasearch of comparing speed of fermentation as well as quality of product. Introduction: Research about Saccharomyces Cerevisiae and Saccharomyces.spp wether which type of yeast has the higher degree of activity, to make a better kind of wine, is the essential demand to raise the fermentation efficiency and quality of product. Purposes of this thesis are:examine developing speed of yeast, alcohol degree and the pH change of grape during fermentation by Saccharomyces.cerevisiae and Saccharomyces spp (yeast on the husky of grape); and compare developing speed, alcohol degree between those two types of yeast at different pH conditions. Materials: 1. Grape Understanding the types of grapes used in wine and their characteristics is vital to make good wine. There are many kinds of grapes to choose from, depending on where you live. There are two main families: Vitis Vinifera - which is popular in Europe - and

Vitis Labrusca - which is native to Canada and the eastern United States. The traditional and most popular type of grape for wine is the Vitis vinifera species. Belows are some of types in Vitis vinifera species:
Auxerrois creates a neutral wine, fruity and soft. It is mainly grown in Luxembourg and Canada.

Barbera is a low-tainnin grape known for its tarry flavor. It is widely planted in California.

Cabernet Franc is used in Bordeaux added in small amounts for flavor. It is mostly used as an additive to blend with other grapes.

Chardonnay is one of the most popular and easiest to grow white grapes. It is grown in Burgundy, California, Australia and South Africa.

Pinotage is developed in the early 1900s and used primarly by South Africa. Pinotage wine is hearty with a fruity and spice taste.

2. Wine yeast Yeast is used in winemaking, where it converts the sugars present in grape juice (or must) into ethanol. Yeast is normally already present on grape skins. Most added wine yeasts are strains of Saccharomyces cerevisiae, though not all strains of the species are suitable. Saccharomyces cerevisiae is a species of yeast. It is perhaps the most useful yeast, having been instrumental to baking and brewingsince ancient times. It is believed that it was originally isolated from the skin of grapes. All strains of S. cerevisiae can grow aerobically on glucose, maltose, and trehalose and fail to grow on lactose and cellobiose. However, growth on other sugars is variable. Galactose and fructose are shown to be two of the best fermenting sugars. The ability of yeasts to use different sugars can differ depending on whether they are grown aerobically or anaerobically. Some strains cannot grow anaerobically on sucrose and trehalose. Saccharomyces spp is a natural yeast. It is believed that Saccharomyces.spp was founded in the skin grape when it is being fermentated.

Saccharomyces spp

Saccharomyces cerevisae

Different Saccharomyces yeasts have differing physiological and fermentative properties, therefore the actual strain of yeast selected can have a direct impact on the finished wine. 3. Sodium bisulfite

While the related compound, sodium metabisulfite, is used in almost all commercial wines to prevent oxidation and preserve flavor, sodium bisulfite is sold by some home winemaking suppliers for the same purpose. In fruit canning, sodium bisulfite is used to prevent browning (caused by oxidation) and to kill microbes. In the case of wine making, sodium bisulfite releases sulfur dioxide gas when added to water or products containing water. The sulfur dioxide kills yeasts, fungi, and bacteria in the grape juice before fermentation. When the sulfur dioxide levels have subsided (about 24 hours), fresh yeast is added for fermentation. It is later added to bottled wine to prevent the formation of vinegar if bacteria are present, and to protect the color, aroma and flavor of the wine from oxidation, which causes browning and other chemical changes. The sulfur dioxide quickly reacts with oxidation by-products and prevents them from causing further deterioration. Methods:

Grapes Taking out the grape skin

Presssing PH= 4.0-4. (Sugar), Citric acid 4

Stir Pasteurizing (NaHSO3 122g/l)

pH = 3.1 4.5 Brix = 220Bx

S.spp (B) Yeast S.cerevisae

Fermentating n

A1 (B1, B2)

A2 (B1, B2) Analysing

A3 (B1, B2)

Grapes were fresh, clean before being take out the skin. After that, grapes were pressed to take the juice. Grape juice was pasteurized by 122mg/l sodium bisulfate (NaHSO3) for 30 minutes. The yeast was added to the sterilized juice. pH and Brix were adjusted by pH meter and Brix meter. We added the yeast again and the mixture war stirred thoroughly. The first sample N0 was analysed. Then, grape juice was dealed to bottles which have 250ml volume. Each bottle was covered to be fermentated. We analysed the sample every 12 hoursto observe the fermentation process. We investigated the effect of pH on the fermentation process for two yeast A1 : pH = 3.1 A2 : pH = 4.1 A3 : pH = 4.5 We also observed the fermentation rate on two yeast B1 : Saccharomyces spp B2: Saccharomycess cerevisae RESULT AND DISCUSSION Compare the development of 2 yeast genera:

1. Effect of pH on the developing speed, Brix degree, Alcohol degree by the time during the fermentation process using Saccharomyces Spp.
Alcohol Degree(%V) Time (hour) Figure 1. Effect of pH on developing speed of yeast by the fermentation time

Time (hour) Figure 1. Effect of pH on developing Alcohol yeast by the fermentation time fermentation time (Counting bacteria by Petri plate)

Time(hour) Figure 2. Effect of pH on degree by the

Time (hour) Figure 3. Effect of pH on degradation speed of developing Brix degree by the fermentation fermentation time

Time(hour) Figure 4. Effect pH to speed of yeast by the (Counting yeast cells by microsope)

2. Effect of pH on the developing speed, Brix degree, Alcohol degree by the time during the fermentation process using Saccharomyces Cerevisiae.
Alcohol Degree (%)

Time(hour)

Time(hour)

Figure 5. Effect of pH on developing Alcohol yeast by the fermentation time time (Counting bacteria by Petri plate)

Figure 6. Effect of pH on degree by the fermentation

Time (hour) Figure 7. Effect of pH on degradation speed developing of Brix degree by the fermentation time fermentation time

Time(hour) Figure 8. Effect pH to speed of yeast by the (Counting yeast cells by microsope)

Follow those figures, we can see: in the first 24 hours of fermentation process, the amount of glucose dropped slowly because the amount of yeast was low. In that time, alcohol formation was little because the yeast cell was focusing on creating the biomass. After 24 hours, developing speed of yeast was very high and strong (developed with exponential function), which made Brix degree reduced strongly. At the same time, Alcohol degree raised very fast. Figure 1 shows us: the maximum developing speed is from 48 to 72 hours, strength of yeast cells was maximum (Nmax: 1010 cfu/ml). The second reason is the amount in the environment was very high, so it inhibited yeast cells. Therefore the alive yeast reduced within 72 hours. At once, the amount of glucose also reduced slowly, so the amount of alcohol raised slowly. Figure 2 shows us the Alcohol degree raised fastly from 48 to 72 hours, then raised slowly because of the low gkucose amount, lots of death yeast cells. The chart about the change of pH within the fermentation process shows the lowest pH at 48th hour, which can be explained by the reason: during the fermentation process, beside alcohol, there was also organic acid. After 48 hours, pH raised because at that time, the amount of glucose was very low, some microorganisms could use organic acid to create energy. After 96 hours, the amount of glucose was very low and pH raised fastly because of that reason. The charts shows the developing speed and alcohol amount of Saccharomyces.sp got the maximum rate at pH 3.7, and Saccharomyces Cerevisiae got the maximum rate at pH 4.1.

3. Compare the development of Saccharomyces.sp and Saccharomyces Cerevisiae at different pH

Time(hour) Time(hour) Figure 9. Developing speed of Saccharomyces.sp between Saccharomyces Cerevisiae at pH 3.7 and Saccharomyces (Counting bacteria by Petri plate) Serevisiae Alcohol degree (%V)

Figure10. Degradation of Brix Saccharomyces.sp

Time(hour) Figure 11. Alcohol degree between Saccharomyces.sp Saccharomyces.sp and Saccharomyces Cerevisiae

Time(hour) Figure 12.Developing speed of Saccharomyces Cerevisiae at pH 3.7 (Counting yeast cells by microsope)

pH Brix degree Amount of yeast cells (Counting by Petri plate)

Amount of yeast cells (Counting by Microscope)

Table 1. Brix degree, developing speed of yeast at different pH by different examining methods

According to the figures and tables, these 2 types of yeast developed slowly in the beginning of fermentation process before hour 24th, then their speed raised fastly from 24 to 72 hours.with exponential function. The amount of yeast cells raised fastly, the amount of glucose reduced rapidly, Alcohol degree and pH reduced rapidly. But the difference between these 2 types of yeast was less than 5% in different conditions. So grape wine fermentated by Saccharomyces cerevisiae and Saccharomyces.sp was different just a little. Conclusion: After the experiment, we have conclusion: _ Developing speed of Saccharomyces.sp and Saccharomyces cerevisiaee was different less than 5% in conditions of oH 3.1, 4.1 and 4.5. The maximum rate of cells reached 1010 cfu/ml. _ Maximum rate of Alcohol degree didnt have diffrence more than 5% at pH 31., 4.1 and 4.5. _ May the genus of yeast in nature: Saccharomyces.sp is genus Saccharomyces.cerevisiaee _ We can use the genus Saccharomyces.sp to make wine instead of Saccharomyces.cerevisiae with the same high quality. Suggestion: Because of the limited time, this thesis could just examine Saccharomyces.sp and Saccharomyces cerevisiaee at different pH conditions. During the examining period, 2 types of yeast didnt show the difference. To have better result, we need: _ Examine effect of Brix degree _ Examine effect of temperature References: 1. Fruit wine making at home Vu Cong Hau, 1993 2. Fruit Microbiology Ho Quang Tri, 1999 3. Microbiology Technology 1, 2, 3 Nguyen Duc Luong University of Technology, 1996
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4. Biotechnology Experiments Nguyen Duc Luong, 2002 5. Microbiology Technology Luong Duc Pham, 1998 6. Fermentation Processing - Tri Nhan Minh, Teshome Edae Jiru, Naznin Sultana, Michael Wawire, 2001

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