Effect of yeast level and fermentation conditions on

physico- chemical composotion of tomato wine

JAGADISH1, K.RAMACHANDRA NAIK2, LAXMAN KUKANOOR3, B.C.PATIL4

AND CHAYA. P.PATIL5

Dept. of Post Harvest Technology, Kittur Rani Channamma College of Horticulture, Arabhavi,
University of Horticultural Sciences, Bagalkot- 587102, India.

Corresponding author- Jagadishhorti@gmail.com

ABSTRACT

An investigation was conducted to study the influence of three yeast levels at

0.20g, 0.25g and 0.30g/l with fermentation conditions (7 days aerobic and 14 days
anerobic fermentation ) by inoculating Saccharomyces ellipsoideus yeast strain for all
the treatments. The physico-chemical properties of fermented tomato juice studied
which includes TSS, pH, titratable acidity, total sugars, reducing, non reducing sugars,
lycopene and ascorbic acid has shown decreasing as ageing progresses upto six months
which is stored under cold storage condition (14 0c). The results of the biochemical
parameters of the prepared tomato wine was evaluated when fresh wine prepared once,
after three and six months of cold storage (14±10 0C). During ageing, most colour
parameters showed higher values in both fermented juices. It was concluded that use of
appropriate yeast level level (0.20g/l) with 14 days anaerobic fermentation gave better
physico-chemical qualities compared to other treatments.

KEYWORDS: Saccharomyces, fermentation, tomato, wine, yeast, aerobic, anaerobic,

storage, levels, conditions

INTRODUCTION

Tomato is a perishable commodity and has to be processed to extend its shelf

life. The well known existing processed forms of tomato include juice, paste, puree,
soup, ketchup, sauce and canned tomatoes (Motamedzadegan and Tabarestani, 2011).
Winemaking could help to broaden the processing avenues of tomato and also increases
the utilisation of wastage of tomato pulp from seed extraction industries.
 The production of any alcoholic beverage involves alcoholic fermentation,
which may start spontaneously either by wild yeast or by inoculation of must or juice
with yeast of known characteristics (Jackson, 2008). The strain of the yeast and the level
of yeast inoculated are among the most important factors in determining the quality of
the alcoholic beverage produced. The use of inappropriate ino-culum levels may lead to
problems such as competition among yeast cells leading to premature death and hence
large quantities of residual sugar, etc. Therefore, it is important that the right level of
inoculum is utilized in fermentation to make maximum use of the available fermentable
sugar. In the present studies, impact of the different yeast level on the physico chemical
composition of wine has been investigated.

MATERIALS AND METHODS

Preparation of must

The tomato was washed thoroughly with tap water. The tomatoes were cut into
smaller pieces with the help of a sharp knife and crushed into fine slurry to separate
seeds and skin, the extract was filtered through a clean muslin cloth to obtain juice.
Table sugar of a concentration of 500 g/L was added to raise the TSS from 5.10 to 24
°Brix. The pH of the mixture was adjusted from 4.82 to 3.20 using citric acid. The dry
yeast used, Saccharomyces ellipsoideus (Angel Yeast Company Limited) was brought
from bangalore and kept in a refrigerator at 5oC according to the manufacturer’s
instructions. The yeast culture was prepared in YDP medium (yeast extract = 0.5%
(w/v), peptone = 1.0% (w/v) and glucose = 2% (w/v)) in a 250-mL Erlenmeyer flask.
The medium was sterilised was suspended in 100 mL of sterilised medium and heated
to 40o for 20 min. to rehydrate the yeast cells . It was then cooled to room temperature
(25o) for adaptation and then it is inoculated to the tomato. The experiment consist of
6 treatments viz., (T1- 0.20g/l + 7 days aerobic fermentation, T2- 0.20g/l + 14 days
anaerobic fermentation T3- 0.25g/l +7 days aerobic fermentation T4-0.25g/l + 14
days anaerobic fermentation T5- 0.30g/l + 7 days aerobic fermentation T 6- 0.30g/l + 14
days anaerobic fermentation) after fermentation which is filtered in bootles allowed for
ageing in cold storage condition.
 48
47

Analysis of fermented tomato juice : The method of Sadler and Murphy (2010) was used
to determine TA and the results were expressed in g/L citric acid. The pH was measured
using a pH meter (PHS-2C Precision pH/mV meter, China) after calibration with
solutions of pH 7 and 4, respectively according to the AOAC (1984). The TSS was
determined by using hand Refractometer and the values expressed in degree brix
(°Brix). The residual sugar content was analysed by DNSA method.

RESULTS AND DISCUSSION

In the present investigation the data on changes in TSS content of tomato

wine as influenced by different yeast levels are presented in Table 1. In fresh wine,
o
the highest TSS (10.29, 9.67 and 9.21 B) was recorded in T2 (0.20g/l + 14 days
o
anaerobic fermentation) While the lowest TSS (9.02, 8.57, 8.06 B) was recorded in
T3 (0.25g/l + 7 days aerobic fermentation) at fresh 3 and 6 months of ageing. The
amount oxygen absorbed also depends on the duration of pulp contact during
maceration (Jackson, 1994). The reduction in the TSS during aging of wines is
expected to be due to the higher yeast activity that may still prevail during aging which
converts sugars into alcohol (Deta et al., 2004).

pH

The data pertaining to pH revealed significant differences among the treatments

4847
(Table 1). The mean pH of fresh wine and aged wines indicated an increase in trend
of pH from the original must pH (3.20) level to end of ageing irrespective of the
treatments imposed . In the fresh wine, the highest pH (3.60, 4.13, 4.62) was noticed
in T2 (0.20g/l + 14 days anaerobic fermentation) which was at par with T5 (3.53).
The lowest pH (3.35, 3.64, 4.23) was recorded in T6 (0.30g/l + 14 days anaerobic
fermentation) respectively. The increase in pH was due to reduction in acidity
through precipitation of potassium tartrate salts from wine or due to enhanced
synthesis of esters from ethyl alcohol and volatile acids. The changes in the pH were
not correlated with the changes in total acidity because of the buffering capacity
of the wines and the relative amount of various acids influencing the acidity
(Shankar et al., 2004)
Total titratable acidity (%)

From the table it is evidenced that there was decrease in trend of titrable acidity
content of wine as the ageing period progressed irrespective of levels of yeast used
and fermentation condition adopted. The results on the total titrable acidity
content (%) of the fresh wine as influenced by the treatments exhibited a wide
range from 1.13 to 1.30 per cent and found significant (Table 2 ). However, the
significantly higher acidity content (1.30, 1.05 and 0.94%) was found in the
treatment T2 (0.20g/l + 14 days anaerobic fermentation) and was found on par with
T4 (1.27). However, the lowest acidity content was associated with T3 (1.13, 0.75
and 0.66%) at fresh , three and 6 months of ageing respectively. The decrease in the
acidity during aging might be due to combination of acids with alcohol to form esters
which adds aroma to the wine during aging (Shankar et al., 2004).

Total sugar (%)

The data pertaining to the total sugar content (%) of tomato wine as influenced by
different yeast levels and fermentation conditions (Table 2) was found significant
and observed in decreasing trend as the ageing period is progressed. Further the mean
total sugars content of wine showed significantly higher total sugars of (12.20, 12.03
and 11.43 per cent) were found in T2 (0.20g/l + 14 days anaerobic fermentation) in
freshly prepared wine and the lowest per cent of total sugars (11.19, 10.82 and
10.22%) was found in treatment T1. This demonstrates difference with 3 different

yeast strains leading to differential activity of yeast and subsequent utilization of

sugars in the respective treatments. The variation in the residual sugars of wine
due to treatment differences have been reported by Somesh et al. (2009) in
strawberry wine; Yadav et al. (2009) in mahuawine.

Reducing sugar (%)

The data on reducing sugars content of tomato wine as influenced by different yeast
levels and fermentation conditions are observed in decreasing trend irrespective of
the treatment combination imposed (Table 3) . The treatment T2 (0.20g/l + 14 days
 51

anaerobic fermentation) exhibited significantly highest level of reducing sugars at

all the stages of estimation viz., fresh wine 8.51 per cent, at 3 and 6 months of ageing
(8.12 and 7.76 per cent respectively) under cold condition. Whereas, the lowest
reducing sugars were recorded in treatment T3 (0.25g/l + 7 days aerobic fermentation)
with 7.39, 6.93 and 6.53 per cent respectively in fresh, 3 and 6 months of ageing.

Non-reducing sugars (%)

The overall decrease in value of non reducing sugar content with the ageing of
tomato wine as irrespective of different yeast levels and fermentation conditions
imposed (Table 3).However, the maximum non-reducing sugars were recorded in T3
(0.25g/l + 7 days aerobic fermentation) with fresh wine (4.53%) as well as at 3 and 6
months of ageing (3.82% and 3.67%) respectively under cold condition and
minimum non reducing sugars were recorded in T1 (3.00, 2.69 and 2.36%) in fresh,
3 and 6 months of ageing under cold condition respectively. Similar trend was
observed with respect to change in the reducing and non reducing sugar level of
tomato fruit wine before and after aging. The low content of residual sugars of litchi
wines indicates that almost all the reducing sugars were consumed during fermentation
(Singh and Preetinder, 2009). The variation in the residual sugars of wine due to
treatment differences have been reported by Somesh et al. (2009) in strawberry wine;
Yadav et al. (2009) in mahua wine; Adsule and Kadam, (1995) in pomegranate wine

51
and Thippesha et al. (1997) in mango wine. Kotecha (2010b) recorded total residual
sugar of 4.30 per cent in pomegranate wine to 7.17 per cent in banana wine whereas,
Bhajipale et al. (1998) in karonda wine recoded a maximum of 12.82 per cent.

Ascorbic acid (mg/100g)

The data pertaining to ascorbic acid (mg/100g) content found in decreasing trend as
the ageing progressed irrespective of yeast levels and fermentation conditions.
revealed significant differences among the treatments (Table 4). However, the
highest ascorbic acid content were recorded in T2 (0.20g/l + 14 days anaerobic
fermentation) in the fresh, 3 and 6 months of ageing (6.68, 6.32 and 6.14 mg/100g)
respectively. The lowest (5.64, 5.41 and 5.28 mg/100g) were recorded in T3 (0.25g/l
+ 7 days aerobic fermentation) in fresh after 3 and 6 months of ageing under
cold condition (14±1 ºC). The reduction in vitamin C content might be due to the
effect of heat during pasteurization. The ascorbic acid also acts as an antioxidant to
help prevent molecular changes caused by oxidation and as a promoter of iron
absorption (Wardlaw, 1999).

Lycopene (mg/100g)

Lycopene content of tomato wine was analysed soon after fermentation and later at 3
and 6 months of ageing in cold condition (14±1 ºC) to know the effect of different
yeast levels and fermentation conditions (Table 4 ).

The highest lycopene (1.78, 1.73 and 1.56 mg/100g) content in the fresh wine was
noticed in treatment T3 (0.25g/l + 7 days aerobic fermentation). Whereas, treatment
T1 (020g/l + 7 days aerobic fermentation) was recorded the lowest lycopene content
(1.42, 1.30 and 1.07 mg/100g) at fresh, three and six months of ageing. Similar
reduction (15.36 to 12.33/100g) of lycopene content in tomato puree was observed that
the reduction mainly due to effect on processing and storage conditions after 6 months
of ageing was obtained by Basappa, (2003)

Conclusion:
Tomato wine which is fermented with yeast level 0.20g/l +14 days anaerpbic
fermentation using saccharomyces ellipsoideus yeast strain showed the significantly
superior with respect to all physiochemical parameters.

REFERENCE

A.O.A.C. (1980) Official methods of Analysis, Association of Official Analytical

Chemists.Washington, D.C. pp. 161-165.

Bhajipale, B. J., Gupta D. N. and Mehta, (1998) Effect of different stages of ripening of
fruit of karonda wine. Indian Food Packer, 54: 27-30.

Basappa, P. B., (2003) Processing of tomato, M.Sc. Thesis, Univ. Agric. Sci., Dhawad
Deta, B. C., Vijay, S., Poonam, S. and Srinivastava, V. K., (2004) Physico-chemical
changes of lime-aonla spiced beverage during storage. J. Food Sci. Technol., 41:
329- 332.

Jackson, R. S., (1994) Wine science: Principles and Application, Academic Press,
pp. 236,191,592.

Jackson SR (2008). Wine Science: Principles and Applications. 3rd ed. Elsevier Inc.
London, pp. 344-345.

Kotecha, P. M., (2010), Possibilities of exploitation of other fruits for wine making,
Compendium of winter school on value addition of grapes with special
reference to wine making at Rahuri (Maharashtra), 5-25 Feb 10, pp. 1-198.

Motamedzadegan A, Tabarestani HS (2011). Tomato Processing, Quality and Nutrition.

In: Sinha NK (ed.) Handbook of Vegetables and Vegetables processing.
Blackwell Publishing Ltd, Iowa, USA. pp. 739-757.

Nigam, J. N., (2000) Continuous ethanol production from pineapple cannery waste
using immobilized yeast cells. J. Biotechnol., 80: 189-193.

Shankar, S., Dilip, J. and Narayana, Y., ( 2004) Changes in chemical composition
of guava wine during storage. Indian Food Packer, 12 : 56-58.

Somesh, S., Joshi, V. K. and Ghanshyam, A., (2009) An overview on strawberry wine
production technology, Composition, maturation and quality evaluation.
Natural Product Radiance, 8 (4): 356-365.

Sadler GD, Murphy PA (2010). Ph and Titratable Acidity. In: Nielsen SS (ed). Food
Analysis. Springer Science + Business Media, LLC, New York, pp. 219-238.

Singh, R. S. and Preetinder Kaur., (2009) Evaluation of litchi juice concentrate for the
production of wine. Natural Product Radiance, 8(4) : 386-391.

Thippesha, D., Chikkasubbanna, V. and Jayaprasad, K. V., (1997) Evaluation of

mango (Mangifera indica L.) varieties for wine making. Beverage and Food World.,
pp. 26-29.
Wardlaw, G.M., (1999) Perspectives in Nutrition, McGraw-Hill, USA p.443.

Yadav, P, Garage N and Diwedi D.H., (2009) Standardization of pre-treatment

conditions for mahua wine preparation. J. Ecofriendly Agric., 4 (1): 88-92.
 Table 1. Effect of ‘Yeast level and fermentation conditions’ on Total soluble solids and pH of fresh and after ageing of tomato
fruit wine under cold storage condition (14 ± 1ºC)

TSS (OBrix) pH

Aging in cold storage Aging in cold storage

Treatments
Fresh wine (months) Fresh wine (months)

3 6 3 6

T1 9.22 8.57 8.06 3.42 3.91 4.32

T2 10.92 9.67 9.21 3.60 4.13 4.62

T3 9.02 8.60 8.18 3.36 3.81 4.44

T4 9.32 9.07 8.45 3.44 3.76 4.42

T5 9.30 8.92 8.37 3.53 3.73 4.23

T6 9.70 9.12 8.65 3.35 3.64 4.31

Mean 9.47 8.98 8.48 3.45 3.83 4.39

S.Em.± 0.06 0.07 0.10 0.04 0.03 0.03

C.D. at 1% 0.26 0.33 0.42 0.18 0.10 0.12

T1- 0.20g/l + 7 days aerobic fermentation T 2- 0.20g/l + 14 days anaerobic fermentation
T3-0.25g/l + 7 days aerobic fermentation T 4- 0.25g/l + 14 days anaerobic fermentation
T5-0.30g/l + 7 days aerobic fermentation T 6- 0.30g/l + 14 days anaerobic fermentation
 Table 2. Effect of ‘Yeast level and fermentation conditions’ on Total titrable acidity and total sugars of fresh and after ageing
of tomato fruit wine under cold storage condition (14 ± 1ºC)

Titratable acidity (%) Total sugars(%)

Aging in cold storage Aging in cold storage

Treatments
Fresh wine (months) Fresh wine (months)

3 6 3 6

T1 1.15 0.92 0.74 11.19 10.82 10.42

T2 1.30 1.05 0.94 12.20 12.03 11.43

T3 1.13 0.83 0.66 11.85 11.74 10.85

T4 1.27 0.75 0.71 11.23 11.14 10.22

T5 1.14 0.89 0.86 12.09 11.86 11.17

T6 1.21 0.77 0.72 12.11 11.64 10.32

Mean 1.20 0.86 0.77 11.77 11.53 10.73

S.Em.± 0.02 0.03 0.04 0.05 0.04 0.04

C.D. at 1% 0.09 0.12 0.15 0.20 0.12 0.15

T1- 0.20g/l + 7 days aerobic fermentation T 2- 0.20g/l + 14 days anaerobic fermentation
T3-0.25g/l + 7 days aerobic fermentation T 4- 0.25g/l + 14 days anaerobic fermentation
T5-0.30g/l + 7 days aerobic fermentation T 6- 0.30g/l + 14 days anaerobic fermentation

Table 3 . Effect of ‘yeast level and fermentation conditions’ on Reducing sugars and Non reducing sugars acidity of fresh and
 after ageing of tomato fruit wine under cold storage condition (14 ± 1ºC)

Reducing sugars (%) Non reducing sugars(%)

Aging in cold storage Aging in cold storage

Treatments
Fresh wine (months) Fresh wine (months)

3 6 3 6

T1 8.12 7.72 7.23 3.00 2.69 2.36

T2 8.51 8.12 7.76 3.77 3.36 3.11

T3 7.39 6.93 6.53 4.53 3.82 3.67

T4 8.27 7.27 6.82 3.07 2.76 2.56

T5 7.93 7.51 7.13 4.24 3.74 3.52

T6 7.87 7.22 6.65 3.27 3.10 2.81

Mean 8.01 7.46 7.02 3.64 3.24 3.00

S.Em.± 0.04 0.03 0.02 0.02 0.03 0.02

C.D. at 1% 0.13 0.15 0.07 0.09 0.12 0.09

T1- 0.20g/l + 7 days aerobic fermentation T 2- 0.20g/l + 14 days anaerobic fermentation
T3-0.25g/l + 7 days aerobic fermentation T 4- 0.25g/l + 14 days anaerobic fermentation
T5-0.30g/l + 7 days aerobic fermentation T 6- 0.30g/l + 14 days anaerobic fermentation

Table 4. Effect of ‘yeast level and fermenntation conditions’ on lycopene and ascorbic acid content of fresh and after ageing of
tomato fruit wine under cold storage condition (14 ± 1ºC)
 Lycopene (mg/100g) Ascorbic acid (mg/100g)

Aging in cold storage Aging in cold storage

Treatments
Fresh wine (months) Fresh wine (months)

3 6 3 6

T1 1.42 1.30 1.07 6.37 6.16 6.09

T2 1.62 1.47 1.36 6.68 6.32 6.14

T3 1.78 1.73 1.56 5.64 5.41 5.28

T4 1.55 1.37 1.28 6.15 6.07 5.56

T5 1.48 1.34 1.21 6.20 5.96 5.48

T6 1.71 1.67 1.42 5.87 5.69 5.36

Mean 1.59 1.48 1.31 6.15 5.93 5.65

S.Em.± 0.02 0.04 0.05 0.07 0.08 0.10

C.D. at 1% 0.07 0.15 0.22 0.30 0.33 0.45

T1- 0.20g/l + 7 days aerobic fermentation T 2- 0.20g/l + 14 days anaerobic fermentation
T3-0.25g/l + 7 days aerobic fermentation T 4- 0.25g/l + 14 days anaerobic fermentation
T5-0.30g/l + 7 days aerobic fermentation T 6- 0.30g/l + 14 days anaerobic fermentation