1. Determination of Optimal
Conditions & Microbial Strains
for Producing Drinking Tomato
Vinegar ..
P.L.S Viduranga
AG/2011/3171
Department of Food
Science
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2. Supervision
Dr: Buddhika Perumpuli
Senior Lecturer in Food Science
Department of Food Science
Faculty of Agriculture
University of Ruhuna
Dr: M. Kaumal
Senior Lecturer
Department of Chemistry
Faculty of Science
University of Colombo
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3. Introduction
 Vinegar is a liquid consisting mainly of acetic acid
(CH3COOH) and water.
 Vinegar is now mainly used as a cooking ingredient.
 Vinegar production is a two stage process,
Sugar Ethanol Vinegar
Bakers Yeast Acetic Acid Bacteria
Alcohol Fermentation Acetic Acid Fermentation
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4. Fruit vinegars are made from fruit wines, usually without
any additional flavoring.
 More recently, vinegars made from various fruits with
distinctive sensory appeal have emerged in the
marketplace.
Traditionally, fruit vinegars were only considered as food
ingredient or as delicate flavor enhancer.
New findings and information suggest that they can also be
regarded as potential functional foods.
Fruit Vinegar
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5.  Tomatoes are the most widely consumed fresh fruit in the
world.
 There is an increasing surplus of tomatoes due to average
production & yield of tomatoes have been steadily
increasing worldwide & sometimes due to their failure to
meet customer standards.
 Therefore a novel processed tomato product with
additional functional enhancements is required for greater
demand for tomato products.
 Due to that, to allow both value additions & the effective
reuse of surplus tomatoes we are going to develop drinking
tomato vinegar.
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6. 6

7. Justification
 Tomatoes are rich in vitamins, trace elements , significant
amount of carotenoids, several polyphenols, rutin &
naringenin.
 During the past decade, average production and yield of
tomatoes have been steadily increasing worldwide.
 Higher wastage of tomatoes can be seen all over the world
due to excess production, poor post harvest handling &
lack of use of available preservation applications.
 Therefore to allow both value additions and the effective
reuse of surplus tomatoes is very important.
 Recently, vinegar obtained from tomatoes is popularizing
as a beverage & as a cooking ingredient. 7

8. Research Objectives
Specific Objective:-
 Determination of the optimal conditions & best microbial
strains for the production of drinking tomato vinegar.
General Objectives:-
 Production of tomato wine at 30oC & 37oC with using
baker’s yeast. (Saccharomyces cerevisiae)
 Production of acetic acid at 30oC & 37oC with using
Acetobacter pasteurianus SL13E-2, SL13E-3, SL13E-4
strains.
 Simultaneous tomato vinegar production with using those
3 strains at best temperature conditions.
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9. Methodology
 Experimental design : Complete Randomized Design
 Strains that will be used,
Baker’s Yeast - Saccharomyces cerevisiae
Acetic Acid Bacteria – Acetobacter pasteurianus
-SL13E-2
-SL13E-3
-SL13E-4
(Strains isolated from Sri Lankan coconut water vinegar.)
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10. 1.Tomato Wine Production
Wash Tomato
Juice
Inoculate by
baker’s yeast
Sugar
10% brix
Incubate at 30oC & 37oC under static conditions
Sampling at every 24 hours
Measure pH, Alcohol, Sugar Content, Acidity
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11. 2.Acetic Acid Production
Potato
Medium
Pre culture at 30oC for
24 hours
YPGD medium +
4 or 6% ethanol
Incubation at 30oC & 37oC
at static culture
Sampling at every 24 hrs
Alcohol %, Acidity % , and pH will be
measured
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Acetate production will be compared with that of thermo-tolerant
strain (Acetobacter pasteurianus SKU 1108)

12. 3.Simultaneous Tomato Vinegar Production
by Dry Yeast and AAB strains
Tomato Juice 40ml (10 % Brix)
+ 0.2 g Dry Yeast
Incubated statically at 30 & 37C
Samples will be collected at every 24 hours to measure acidity.
Alcohol, sugar and pH
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Final selected product will be tested for antioxidant availability.

13. 4.Analytical Methods
 Sugar- Brix Meter
- HPLC Analysis
 Alcohol- HPLC Analysis
 Acidity- Titration with NaOH
 pH- pH Meter
 Antioxidant- DPPH Radical Scavenging
 Data will be analyzed by using descriptive methods and
inferential methods.
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14. Time Frame
Activity Month
July August September October November December
1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4
Literature survey
Research Proposal
Preparation of the
tomato juice
Inoculation of yeast to
the juice & incubation
Pre culture of 3
microbial strains
Production of vinegar by
using simultaneous
fermentation of both
yeast & acetic acid
bacteria
Final presentation
Thesis writing
Thesis submission 14

15. References
Anon, (2015). [online] Available at:
http://file:///E:/My%20research/The%20Health%20Care%20Effect%20of%20Fruit%20Vi
negar%20-%20Chempedia%20-%20LookChem.html [Accessed 17 Jul. 2015].
Continuous microbiological production of acetic acid and vinegar. (1988). Environment
International, 14(2), p.III-IV. [online] 12(2), pp.127-138. Available at:
http://creativecommons.org/licenses/by/4.0[Accessed 22 Jul. 2015].
Krusong, W. and Vichitraka, A. (2010). An investigation of simultaneous pineapple
vinegar fermentation interaction between acetic bacteria and yeast. Asian journal of
food and agro chemistry, [online] 3(1), pp.192-203. Available at: http://www.ajofai.info
[Accessed 17 Jul. 2015].
Lee, J., Cho, H. and Lee, M. (2013). New vinegar produced by tomato suppresses
adipocyte differentiation and fat accumulation in 3T3-L1 cells and obese of rats. Food
Chemistry, [online] (141), pp.3241-3249. Available at:
http://dx.doi.org/10.1016/j.foodchem. [Accessed 18 Jul. 2015].
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16. MALDONADO, O., ROLZ, C. and CABRERA, S. (1975). WINE AND VINEGAR PRODUCTION
FROM TROPICAL FRUITS. Journal of Food Science, 40(2), pp.262-265. , [online] 3(1),
pp.77-123. Available at: http://www.ajofai.info [Accessed 17 Jul. 2015].
Patil, K. (2011). Microbial Production of Vinegar (Sour wine) by using Various Fruits. IJAR,
[online] 3(8), pp.602-604. Available at:
http://creativecommons.org/licenses/by/4.0[Accessed 21 Jul. 2015].
Perumpuli, P., Watanabe, T. and Toyama, H. (2014). Identification and characterization of
thermotolerant acetic acid bacteria strains isolated from coconut water vinegar in sri
lanka. Bioscince,Biotechnology & Biochemistry, [online] 78(3), pp.533-541. Available at:
http://dx.doi.org?10.1080/09168451.2014.882758 [Accessed 16 Jul. 2015].
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17. Thank You!
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