Chapter 1

Introduction

Winemaking technologies are among the oldest known to humans. The existence of wine

is said to be more than 7,500 years, indicated by archaeological excavations that have uncovered

many sites with sunken jars (Kosseva, Joshi & Panesar, 2016). Winemaking is also one of the

oldest techniques known to civilization and is one of the most commercially flourishing

biotechnological processes, even today (Joshi et al., 2011), involving alcoholic fermentation

(Molinos et al., 2016; Joshi, 2016). Moreover, the consumption of red wine dates back thousands

of years to the Egyptians, who prepared medicinal wines made from grapes, using herbs and tree

resins (McGovern & Mirzoian, 2009). Of all the traditional wines, red wine is often advertised as

being very beneficial for human health for being very rich in polyphenols and many other health-

promoting compounds. Grapes are basically the main ingredient in making wines; and it is the

most known. However, grapes are not the only fruit that can be made into wine. Fruit wines can

be made from nearly any fruit, but most popular fruit sources include: blueberries, blackberries,

strawberries, cherries, apples, peaches, and plums (Jagtap & Bapat, 2015).

Juice conversion into wine is a complex biochemical process involving yeast,

predominantly Saccharomyces cerevisiae. In short, “winemaking is the result of a number of

biochemical transformations brought about by the action of several enzymes from various

microorganisms, especially yeast, which carry out the major part of the process of alcoholic

fermentation” (Moreno-Arribas and Polo, 2008). Furthermore, lactic acid bacteria also contribute

high-acid wines (Bartowsky, 2011). To add more, a number of enzymes originating from the

1
fruit, yeast, and lactic acid bacteria or those from contaminating microorganisms are involved in

different steps of winemaking.

In yeast-based alcoholic fermentation, the yeast utilizes the sugars and other constituents

of grape juice, or other fruit juice, for its growth, converting them into ethanol, carbon dioxide,

and other metabolites (Rebordinos et al., 2011). All the products formed during the process

contribute to the sensory quality characteristics and chemical composition of wine. So, being the

fundamental in winemaking, a thorough knowledge of yeast growth and the chemistry of

fermentation involved are very essential. Wine preparation is a traditional process, yet several

advancements have been made on various facets of the process. However, the advances made in

the second half of the 20th century have clearly shown that fermentation of fruit must and the

production of quality wines are not quite as simple (Moreno-Arribas & Polo, 2008). Moreover, it

is relevant to state that the preparation of non-grape wine is basically similar to that from grapes.

Hence, the chemistry of alcoholic fermentation or malolactic fermentation, and associated

characteristics would remain fundamentally the same. However, differences in several

components would still be present because of the difference in the composition of the specific

fruits used.

Additionally, sugars in wine are at the heart of what makes winemaking possible. During

the process of fermentation, sugars from wine grapes are broken down and converted

by yeast into alcohol (ethanol) and carbon dioxide. Typically, wine consists of 10-12% ethanol

produced by natural alcoholic fermentation (Many, Radhika & Ganesan, 2014). No literature

has been found about the study of the optimum sugar concentration in the media for the

fermentation of tomato for wine production.

2
 The purpose of the study is mainly to provide information about the effects of sugar in the

kinetics of tomato wine production that will give a hand in the wine industry. It aims to widen

the knowledge and improve the skills of winemakers to make better quality wines out of

tomatoes. It also aims to give information for wine consumers that will help them to know more

about tomato wines and select the wines of their choice.

This study, “Study on the Effect of Sugar Concentration in the Kinetics of Tomato Wine

Production” will be a significant asset that will help winemakers, wine consumers, as well as

future researchers.

To the winemakers, this study will serve as a guide that will help them know how to

create a tomato wine of great taste and sweetness, their desired alcohol yield, and manipulate

sugar to produce so. This study will also save them some time and resources for they will no

longer need to try making tomato wines, trying to find out the desired sugar and amount for

achieving the desired sweetness and alcohol content produced.

To the wine consumers, this study will give them information about tomato wines and be

aware of what are the effects of sugar to the sweetness and alcohol content of the wine. With

that, they will be more knowledgeable about tomato wines and will actually make them

comfortable upon drinking so. Also, this study will help them figure out the tomato wines of

their choice.

To the future researchers, this study will serve as their guide and reference when taking

this field. They will be guided with this study and come up with the future results that will be of

a great help for the next generation of researchers. The researchers of this study will be very

satisfied for any new data and innovation that the future researchers will gather.

3
 Furthermore, in 2014, Many, Radhika, and Ganesan conducted a study on tomato wine

production and optimization. They investigated the suitability of tomato fruit for the production

of wine. Their study focused on the production of tomato wine with standardized reference to

different variables such as: size of inoculums, pH of tomato juice, and time of fermentation (1-10

days), brix adjustment, incubation, temperature, effect of alcohol adapted and non-adapted

cultures, and age of culture. Since there is no study about the optimum concentration of

sugar in the true media in tomato wine making that has been found, the observation of the

final yield of pH, alcohol, sugar, Total Soluble Solids (TSS), and total cell concentration

after fermentation will be a significant asset in knowing the effects of sugar in tomato wine;

and, the observation of cell growth and concentration in the tomato wine, with time, will

yield to the kinetics of fermentation.

Additionally, baker’s yeast will be used for juice conversion and only one kind of tomato

will be used for fermentation and wine production. This study focuses only on tomatoes that are

grown in the Philippines. Moreover, the experiments will be conducted in the laboratories and

facilities of Mapúa University. Significantly, this study will only use parameters, such as the pH

level of the tomato juice, brix (sugar content), Total Soluble Solids, ethanol content, and cell

growth in determining the chemical and physical properties of the tomato for wine production.

To add more, the study will not compare the produced tomato wine to the properties of grape

wine and other kinds of wine.

4
 Chapter 2

REVIEW OF LITERATURE

For further explorations on this study, the researchers conducted researches about studies

that focused on wines in different perspectives. This chapter will serve as a comprehensive

review of literature of the previous researches pertaining to tomato, wine fermentation, wine-

making, and the chemical processes or the kinetic processes of tomato wine fermentation. By

definition, study on chemical kinetics is the study of chemical processes and rates of reactions

that includes: analysis of conditions that affect speed of a chemical reaction, understanding

reaction mechanisms and transition states, and forming mathematical models to predict and

describe a chemical reaction (Helmenstine, 2017).

2.1 Fundamentals of Winemaking

Hands and Hughes (2001) stated that the fundamentals of winemaking have stayed the

same since biblical times. What has changed is the winemakers’ ability to maintain the sterile

environment required to produce wines of best qualities (Halliday & Johnson, 1994) (Conradie,

Sigge & Cloete, 2014). In addition to that, Myers (2014) refers to wine making as a natural

process that requires little human intervention; and each wine maker escorts the process through

different techniques.

2.2 Properties of Tomato

The property of high soluble solids will cause the advancements of the characteristics of

the final products desired (Moelants et al., 2013). According to Wu et al. (2018), the total feature

and acceptability of the tomato product shall depend greatly in the consistency (such as

viscosity). In relation to that, factors like variety of tomato, growing conditions, and the maturity

of the fruit used can greatly affect viscosity. Moreover, color and flavor are also important

5
attributes in determing the overall quality and acceptability of the tomato product. Huang, Lu,

Hu, and Chen (2018) added that, in order for a tomato to be in a good condition, it should have a

good appearance, both in color and shape, desired surface, aptness flavor, and aroma.

Furthermore, Biswas, East, Hewett, and Heyes (2014) identified that the textural assets of a

tomato often varied by the low temperature storeroom, resulting in remarkable changes in its

firmness.

Additionally, Valdivia-Nájar, Martín-Belloso, and Soliva-Fortuny (2018) studied the

changes of the properties of tomatoes as affected by light treatments and storage time. In their

study, they indicated that the presence of light and oxygen causes the physiochemical reactions

of the substances of bioactive compounds and anti-oxidant potential of a fresh-cut tomato

(Odriozola-Serrano et al., 2008a, 2018b); and these reactions affect the overall quality of a

tomato. In relation to that, they have introduced the Pulse light (PL) treatment that aims to

minimize the growth of microbial properties on the compounds of tomato. They have concluded

that PL treatmeat is an effective treatment in minimizing the growth of microorganisms on the

surfaces of food; though, have negative impact on the antioxidant properties within tomato slices

or insides of the fruit. It was then suggested that PL treatment should be evaluated and optimized

to be able to retain better antioxidant ability and bioactive components of tomato slices, and

preserve the natural properties of the tomato.

2.3 Wine Fermentation

Fermentation is the conversion of sugar into acids and alcohol acid (Helmenstine, 2017).

Malolactic fermentation or MLF is a substantial process in wine production that impacts on

operation efficiency and product quality and safety. MLF is also responsible in reducing wine’s

acidity, stabilizing wine through removal of a potential carbon source, and producing wine’s

6
aroma and flavor changes. The main bacterium responsible for conducting MLF in red, white,

and sparkling base wines is Oenococcus oeni- member of lactic acid bacteria family (Bartowsky,

2014). At its most basic level, organic wine is made, most commonly from grapes that have been

grown with as little human impact as possible

Additionally, water, sugar, or both, may be added within a limited amount for the

production of agricultural wine. In wine fermentation, sugars are transformed to alcohol

according to the variety and ripeness of the crop used; and this leads to the most significant

difference between grape juice and wine (Conradie et al., 2014). According to Many, Radhika,

and Ganesan (2014), wine is a typical alcoholic beverage produced by natural alcoholic

fermentation of fruit juice consisting of 10-12% of ethanol. During winemaking process,

additional compounds are formed and some of those compounds have to be removed before the

bottling process (Conradie et al., 2014).

2.4 Basic Components of Winemaking

According to Myers (2014), harvesting, crushing and pressing, fermentation, clarification,

and aging and bottling are the five basic components of the winemaking process. Harvesting is

an important part of ensuring a delicious wine. It determines the acidity, sweetness, and flavor of

the wine; and it can be done by hand or mechanically. Crushing and pressing stage separates the

juice from the skins, seeds, and solids of the main ingredient. Fermentation is the process that

converts sugar into alcohol; and fermentation of wine usually takes place from 10 days to one

month or more. Clarification is the process in which solids such as dead yeast cells, tannins, and

proteins are removed; and it can be done through fining or filtration. Lastly, aging and bottling

stage is the one responsible for producing smoother, rounder, and more vanilla flavored wine. It

also increases wine’s exposure to oxygen while it ages, which decreases tannin and helps the

7
wine reach its optimal fruitiness. After aging, wines are bottled with either a cork or a screw cap,

depending on the wine maker’s preference. Furthermore, Myers (2014) stated that wine makers

typically follow these five steps but add variations along the way to make their wine unique.

8
Figure 1: Complete Winemaking Process

HARVEST

DESTEMMING WASTE

WHITE WINE CRUSHING RED WINE

WASTE PRESSING FERMENTATION

WASTE
& MACERATION

FERMENTATION
PRESSING WASTE

WASTE SEDIMENTATION
DECANTING COMPLETION OF
FERMENTATION

SEDIMENTATION WASTE
DECANTING

MALOLACTIC FERMENTATION
(IF DESIRED)

SEDIMENTATION DECANTING WASTE

MATURATION AND NATURAL

CLARIFICATION

FINING AND STABILISATION WASTE

BOTTLING

9
2.5 Factors Affecting Fermentation of Fruits

Factors that affect the malolactic fermentation of fruit include temperature, pH, acidity,

ethanol, sulfite and availability of nutrients; which are all important for the growth and metabolic

activities of the lactic acid bacteria. The lactic acid bacteria are more fastidious in their growth

requirements than the yeast and generally require higher temperatures than the yeast in order to

grow. The temperature should be above 18°C (64°F) to allow growth of the bacteria. The

bacteria will grow more rapidly at higher temperatures but may lead to deterioration of the wine.

Moreover, successful malolactic conversions have been reported to occur at lower temperatures,

indicating that some strains can develop temperature tolerance; though this is not a general

phenomenon. Moreover, the pH will affect which strains and species can grow in the wine or

juice and will influence growth rate as well as metabolic activities of the organisms (Amachi &

Yoshizumi, 2014).

2.5 Factors Affecting Alcohol Content during Fermentation

Jang, Woo, Jo, Kim, and Co (2010) investigated the alcohol fermentation conditions of

tomato wine. For semi-mature and full-mature tomatoes, 11% of alcohol content of tomato wine

was prepared; and 10.6% for the immature tomatoes. During the examination of tomato alcohol

fermentation on pre-treatment conditions, the wine from full mature-crushed whole tomato

contained lower alcohol at 10.6% than those of full mature-crushed tomato removed stem and

full mature-tomato juice removed stem containing about 11%. Furthermore, the alcohol content

and sugar content of tomato wine increased in proportion to the concentration of initial sugar

(8.8~14.4% and 5.5~9.2°Brix). Their study found that initial titratable acidity and pH did not

affect the alcohol contents during fermentation; and maltose showed the highest level of 264.6

mg% in the free sugar analysis of tomato wine.

10
 Chapter 3

Study on the Effects of Substrate Concentration in the Kinetics of Tomato Wine

Production

Abstract

Winemaking technologies are among the oldest known to civilization and also one of the
most commercially prosperous biotechnological processes that involves alcoholic fermentation.
It is the result of a number of biochemical transformations brought by several enzymes from
various microorganisms, especially yeast, which carry out the major part of alcohol fermentation
processes. The purpose of this study is to provide information about the effects of sugar in the
kinetics of tomato wine production that will give a hand in the wine industry. Additionally, this
study will only use pH, brix (sugar content), Total Soluble Solids (TSS), and alcohol content
parameters. To add more, this study will observe cell growth with time for the kinetics of
fermentation. This study will not compare the produced tomato wine to the properties of other
kinds of wine. Furthermore, this study followed a quantitative research design to highlight the
measurements of variables and data via statistical, numerical, and mathematical analysis of data
through experimentation. The experiments for this study will be conducted at Mapúa
University’s laboratories and facilities. Moreover, the fermentation kinetics of tomato wine will
be determined and compared among three different batches of tomato wine.

Keywords: Winemaking, Fermentation, Yeast, Kinetics, Tomato

1. Introduction

Winemaking technologies are among the oldest known to humans and is said to have

existed for more than 7,500 years, indicated by archaeological excavations that have uncovered

many sites with sunken jars (Joshi & Kosseva, 2017). Winemaking is also one of the oldest

techniques known to civilization and is one of the most commercially prosperous

biotechnological processes, even today (Joshi et al., 2011), that involves alcoholic fermentation

(Molinos et al., 2016; Joshi, 2016). Moreover, the consumption of red wine dates back thousands

of years to the Egyptians who were used in preparing medicinal wines made from grapes, using

herbs and tree resins (McGovern et al., 2009). Of all the traditional wines, red wine is often

11
advertised as being very beneficial for human health for being very rich in polyphenols and

many other health-promoting compounds. Basically, grape is the main ingredient in making

wines; and it is the most known; however, grapes are not the only fruit that can be made into

wine. According to Jagtap and Bapat (2015), fruit wines can be made from nearly any fruit but

most popular fruit sources include: blueberries, blackberries, strawberries, cherries, apples,

peaches, and plums.

Since there is no study about the optimum concentration of sugar in the true media in tomato

wine making that has been found, the observation of the final yield of pH, alcohol, sugar, total

soluble solids, and total cell concentration after fermentation will be a significant asset in

knowing the effects of sugar in tomato wine; and the observation of cell growth and

concentration in the tomato wine with time will yield to the kinetics of fermentation.

Additionally, baker’s yeast will be used for juice conversion and only one kind of tomato will be

used for fermentation and wine production. Moreover, the experiments will be conducted in the

laboratories and facilities of Mapúa University. This study will only use parameters, such as the

pH level of the tomato juice, brix (sugar content), TSS, ethanol content, and cell growth in

determining the chemical and physical properties of the tomato for wine making. To add more,

the study will not compare the produced tomato wine to the properties of other kinds of wine.

2. Methodology

This study followed a quantitative research design. According to Babbie (2010), a

quantitative approach is used to highlight the measurements of variables and data via statistical,

numerical, and mathematical analysis of data through experimentation. The experiments for this

study will be conducted at Mapúa University’s laboratories and facilities. In this study, the

12
fermentation kinetics of tomato wine will be determined and compared among three different

batches as shown in table 1.

Table 1: Experimental Design

15 ºBrix 22 ºBrix 22 ºBrix

(Control)
Initial Conditions With Pulp With Pulp Without Pulp

Initial pH - - -

Initial Sugar 15 22 22

Initial Total Soluble Observed Density 1 Observed Density 2 Observed Density 3

Solids (TSS)
Kinetics of Fermentation (Cell Growth)

Cell Observed Density 1 Observed Density 2 Observed Density 3

Sample 1 Sample 1 Sample 1

Day 1 Sample 2 Sample 2 Sample 2

Sample 1 Sample 1 Sample 1

Day 2
Sample 2 Sample 2 Sample 2

Sample 1 Sample 1 Sample 1

Day 3
Sample 2 Sample 2 Sample 2

Sample 1 Sample 1 Sample 1

Day 4
Sample 2 Sample 2 Sample 2

Sample 1 Sample 1 Sample 1

Day 5
Sample 2 Sample 2 Sample 2

15 ºBrix 22ºBrix 22 ºBrix

(Control)
Final Yield With Pulp With Pulp Without Pulp

pH - - -

13
 Sugar - - -

TSS - - -

Alchol (Ethanol) - - -
Content

The tomato wine will be made by the researchers, themselves, by following the basic

steps in making tomato wine. The fermentation will take place for 10-30 days and the wine will

then be divided into three different batches and prepared in three different set-ups. The first batch

of tomato wine will be at 15 ºBrix, with pulp; the second batch will be at 22ºBrix, with pulp; and

the third batch will be at 22ºBrix, without pulp. The initial pH, and Total Soluble Solids (TSS)

will then be observed; and, with the second batch as the experiment’s control.

Furthermore, for the fermentation kinetics of tomato wine, cell growth will be obeserved

with respect to time. Cell growth will be observed by taking 2 samples each day, for five days,

on each batch of the tomato wine. The first sample will be taken at 7:00 AM and the second

sample at 5:00 PM. After five days of observation, the final yield of pH, sugar, Total Soluble

Solids, and alcohol (ethanol) will then be observed. With that, the effects of the sugar

concentration in the kinetics of tomato wine will be determined.

2. Materials and Methods

Ripe roma tomatoes will be purchased from local market and will be washed in tap water.

The tomato will be cut into pieces and will be placed into a juicer to separate the pulp. The pulp

will be strained through a tea strainer until 500 mL of juice is collected; and the pH and Totatl

Soluble Solids will be measured. The juice will be adjusted to 22⁰Brix by adding sugar (275 g/L)

(batch 2, control). Then, required volume of juice will be dispensed into boiling tubes and

pasteurised at 80⁰C for 4-5 minutes. Another juice will be adjusted to 15⁰Brix (batch 1) and

14
 another juice to 22⁰Brix with no pulp (batch 3). Figure 1 shows what the experimental set-up

look like and how samples will be taken and observed for data collection.

Figure 1: Experimental Set-up

Syringe 100 mL
(Sampling Port)

Glass Tubing
Rubber Tubing
Rubber Cork

Glass Tubing

5 gal/ 20 L
Plastic Bottle
Tomato Wine

3-L Bottle
(Sterilized
)
3-6 %
NaOH
Solution

The instruments that will be used for the experiment and fermentation processes are:

juicer for pulp and juice separation, strainer for juice collection, pH meter, refractometer for

measuring Brix and TSS, hydrometer for measuring alohol content, microscope for observing

cell growth, three 5 galons plastic bottles that will serve as containers for tomato wine, three

glass tubes, three rubber tubes, three rubber corks, three 3-L sterilized bottle that will serve as

15
containers for NaOH solutions that will also serve as carbon dioxide (CO2) collector, and 100-

mL syringe that will serve as sampling port.

References

Babbie, E., & Benaquisto, L. (2010). Fundamentals of Social Research (Second Canadian
Edition). Toronto, ON: Nelson.
Wine making recipe for tomato wine (n.d.). How to make tomato wine: Winemaking guides
something to try. Retrieved from https://www.pinterest.ph/pin/81627811967049469.

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