Sevilla, Regine Louise Victoria C.

| BSFT 1-1
FTEC 50, January 2022

Raisin Vinegar Production: A Microbiological Perspective

ABSTRACT

Vinegar is a living ingredient created through the process of fermentation. The term
“vinegar” actually refers to the two-step process of fermentation from a carbohydrate to
an alcohol to an acetic acid. Various microorganisms such as yeasts and acetic acid
bacteria are responsible for the conversion of sugar into alcohol, which is subsequently
fermented into vinegar. Both alcoholic and acetic fermentation processes have
considerable impact on the final qualities of the vinegar produced. In this review, a
description of homemade vinegar fermentation process and its implications are presented
along with the microbial activities involved.

Keywords: vinegar, acetic acid bacteria, yeast, fermentation, microbial activities

1.0 INTRODUCTION

A technique for preserving foods and drinks that dates back to the ancient times, far
before refrigeration— fermenting. The interplay of microorganisms at work, as well as the
ingredients and ambient circumstances, are a few of the factors involved in fermentation.
Each of these elements contributes to hundreds of distinct fermented food varieties with
nutritional value. Upon a strictly biochemical point of view, certain microorganisms are
held responsible for the conversion of carbohydrates into alcohol and lactic acids which
makes up the whole process of fermentation. The activity of the microbes would serve as a
natural preservative that can even improve the taste, texture, and nutritional content of the
product.
Spontaneous fermentation was first discovered by accident then by intention. It takes
place when naturally present microbes initiate fermentation with wild yeasts rather than
cultivated ones such as the brewer’s yeast. With this in mind, I decided to ferment vinegar.
In a liquid foundation created from pressed fruit, grains, and cereals, vinegar
fermentation can be spontaneous or controlled by adding a "mother" or certain strains of
bacteria. Furthermore, because vinegar may be made from wine, the antecedents of vinegar
are hard to distinguish from those of wine. Vinegar is a sour-tasting liquid that contains
acetic acid, derived from the French word Vin (wine) and Aigre (sour). Throughout history,
beginning with the discovery of alcoholic fermentation from fruits and vegetables in
agriculture, vinegar has been used as a condiment and a preservative. This paper intends to
address the process and microbiological activities involved in the fermenting vinegar.
________________________

2.0 MATERIALS & METHODS

The basic requirement for vinegar production is a raw material that will undergo an
alcoholic fermentation. In my case, I acquired some naturally sweet and high in sugar Sun-
maid raisins (100g) from the nearest local store. The container used was a 0.41L jar filled
to the neck with mineralized water mixed with an inch of raisins, placed on top of a shelf
with an approximate room temperature of 20-22 °C. A piece of cotton cloth was used to
cover the container tightened by some rubber bands.

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 Sevilla, Regine Louise Victoria C. | BSFT 1-1
FTEC 50, January 2022

2.1 Raisin Vinegar Production

The fermentation process was documented and carried out for a total of 33 days with noted
observations. Some of the angled shots taken shows the top view mouth of the jar and its
side for a clear view of the status of the raisins and color of the water.

Documentation & observations

Week 1

December 7, 2021 – Start of fermenting

process

December 8, 2021 – Day 1

December 9, 2021 – Day 2

• Water was now reddish in
color
• Raisins are starting to get
plump

December 10, 2021 – Day 3

December 11, 2021 – Day 4

• Raisins are now floating
• Some sort of small dry gray
bubbles is formed

December 12, 2021 – Day 5

• Bubbles are now white and
some increased in size
• Faint smell

December 13, 2021 – Day 6

December 14, 2021 – Day 7

• Prominent bubbling

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 Sevilla, Regine Louise Victoria C. | BSFT 1-1
FTEC 50, January 2022

Week 2

December 15, 2021 – Day 8

• Emits a strong smell of alcohol

December 16, 2021 – Day 9

December 17, 2021 – Day 10

December 18, 2021 – Day 11

• Bubbles seem to lessen

December 19, 2021 – Day 12

December 20, 2021 – Day 13

December 21, 2021 – Day 14

• Water looks slightly yellowish
• Raisins are starting to sink

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 Sevilla, Regine Louise Victoria C. | BSFT 1-1
FTEC 50, January 2022

Week 3

December 22, 2021 – Day 15

• No bubbles were formed
December 23, 2021 – Day 16
• Thin membrane-like white
stuff started to form at the sides
• Water started to look murky

December 24, 2021 – Day 17

• Raisins have completely settled
at the bottom

December 25, 2022 – Day 18

December 26, 2021 – Day 19

• White stuffs were increasing;
attempted to remove some by
scooping

December 27, 2021 – Day 20

December 28, 2021 – Day 21

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 Sevilla, Regine Louise Victoria C. | BSFT 1-1
FTEC 50, January 2022

Week 4

December 29, 2021 – Day 22

• Raisins look dissolved
• A streak of white stuff has
formed at the center

December 30, 2022 – Day 23

December 31, 2021 – Day 24

January 1, 2022 – Day 25

• A white cotton-like sheath
appeared on top of the surface
• The water looked clear and
clean after removing the sheath

January 2, 2022 – Day 26

• Water looked a bit cloudy

January 3, 2022 – Day 27

January 4, 2022 – Day 28

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 Sevilla, Regine Louise Victoria C. | BSFT 1-1
FTEC 50, January 2022

Final Days of Fermentation

January 5, 2022 – Day 29

• Water has a thick color

January 6, 2022 – Day 30

January 7, 2022 – Day 31

January 8, 2022 – Day 32

• Another sheath appeared, this
time a lot thicker
• Water looked richer and
thicker

January 9, 2022 – Final Product

2.2 Implications
Preparation of the raw material is one of the crucial steps in vinegar production since this
where fermentable sugar and juice solution are acquired to be acetified. Raisins were used
in this process, demonstrating the importance of basic safe food handling, storage, and
processing to prevent the growth of pathogenic microorganisms. It's vital to maintain your
vinegar in a clean, sterilized container during the fermenting process and the water used
should be pure or filtered because chlorine and fluoride contained in municipal water might
cause detrimental influences. Essentially, a cotton cloth secured by a rubber band is
employed to ensure that it is breathable while keeping out any undesired dust because some
bacteria that aid in the fermentation process require oxygen to function.
As the vinegar ferments, it naturally becomes cloudy. The white stuff mentioned earlier is
called a SCOBY, it is basically a mother. The vinegar mother ranges from a transparent to
dark color with a delicate leaf-like appearance to a solid mass floating on top of the
container. The microbes tend to be temperature and UV light sensitive therefore making it
important to store it at room temperature in a dark spot. Aging the vinegar would be the
final step in the process. This allows esters to mature and the flavors to develop, simply
storing it for several months in a sealed, airtight container will do.

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 Sevilla, Regine Louise Victoria C. | BSFT 1-1
FTEC 50, January 2022

Summary of raisin vinegar fermentation

Week Notable Changes
1 Starts bubbling
2 Pungent smell

3 SCOBY (symbiotic culture of bacteria and yeast) is formed

4 Vinegar “mother” appears

The fermentation process should have concluded on the 30th day, according to custom.
The three-day prolongation was triggered by an attempt to establish another "mother."

3.0 MICROBIAL ACTIVITIES INVOLVED

The word vinegar, according to the Codex Alimentarius (1987), refers to a liquid suited
for human consumption made from a suitable agricultural raw material including starch,
sugars, or starch and sugars, by a twofold fermentation process, first alcoholic and then
acetous. The alcoholic and acetification processes play a crucial role in the manufacturing
of vinegar after the raw material processing. Yeasts are the most common microorganisms
responsible for alcoholic fermentation, whereas acetic acid bacteria are required for
acetification.

Summary of technological steps in vinegar production

Substrate: Raisins (grapes)
Step 1 Step 2

Ethanol Production Acetic Oxidation

Yeasts (alcoholic Acetic acid bacteria (AAB)
fermentation)

3.1 Yeasts
In the first stage, yeast convert sugars into ethanol anaerobically. Yeasts are vegetative
fungi that reproduce mostly by budding or fission, resulting in growth that primarily
consists of single cells. Wild yeast is used in spontaneous fermentation; it is any yeast
variation that occurs spontaneously everywhere, especially on fruits.
The presence of a yeast that belongs to the Class Saccharomycetes; Phylum Ascomycota
(Saccharomyces cerevisiae) converts the sugar of the substrate into ethanol which develops
small bubbles or foaming that merely indicates the generating process of the said
conversion. Being able to ferment efficiently in almost all fermentative substrates, S.
cerevisiae is the most important yeast in the fermentation industry that is believed to have
been originally isolated from the skin of grapes.

3.2 Acetic Acid Bacteria

When alcoholic liquid is exposed to oxygen, AAB starts their growth on the surface. AAB
are present in the environment and in the raw material yet cannot grow during the alcoholic
fermentation because of the anerobic conditions. The second phase, acetification, is where

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 Sevilla, Regine Louise Victoria C. | BSFT 1-1
FTEC 50, January 2022

ethanol is oxidized to acetic (ethanoic) acid aerobically by bacteria of the genera

Acetobacter and Gluconobacter. AAB are gram-negative that show strict aerobic
metabolism with oxygen as catalase positive and oxidase negative. Gluconobacter is
distinguished from Acetobacter by their ability to oxidize acetic acid to carbon dioxide and
water. Acetaldehyde is an intermediate step on the way from alcohol to vinegar which
creates the pungent odor.
Gluconacetobacter species (G. xylinus) generally produce the water-soluble brown pigment
and are motile. Gluconobacter oxydans was the main representative of acetic acid bacteria
on sound, unspoiled red or white grapes. However, A. aceti and, to a lesser extent, A.
pasteiurianius became more prevalent as the grapes became spoiled since Acetobacter spp.
are the better acid producers and are more common in commercial vinegar production.

3.3 Vinegar Mother

Regarding the “mother of vinegar,” it is a thin film that covers the surface of the liquid was
made by “globulles” six times smaller than yeasts; Kützing can be credited with the first
microscopic observation of acetic acid bacteria in 1837. Finally, Pasteur in 1864 claimed
that the transformation of wine into vinegar was due to the development of the veil of
Mycoderma aceti on its surface.
It is completely harmless, mainly composed of cellulose, a byproduct of the actions of
bacteria and yeast that converts alcohol to vinegar, and the presence of one in your vinegar
simply signifies that certain sugars or alcohol weren't entirely fermented throughout the
vinegar process.

4.0 CONCLUSION

Overall, vinegar is an aqueous solution of acetic acid and other constituents, known and
consumed worldwide as a condiment and a preservative. The said product is the result of a
two-stage fermentation. The first step is an anaerobic fermentation (alcoholic fermentation
of sugars into ethanol by yeasts, mainly Saccharomyces cerevisiae). This causes the
occurrence of foaming which simply means that the conversion of sugar is already
generating. The second step is an aerobic fermentation (oxidation of ethanol into acetic acid
by AAB by Acetobacter aceti) which also causes the strong smell of alcohol by producing
acetaldehyde. Unfiltered homemade vinegar may develop a film (Mycoderma aceti) that
can make the vinegar cloudy, it is nothing to be feared.

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 Sevilla, Regine Louise Victoria C. | BSFT 1-1
FTEC 50, January 2022

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