(i)

CHEMICAL PROCESS LABORATORY 1

EXPERIMENT NO. 4
FERMENTATION OF PINEAPPLE WINE

Charlagne Aranzo
Jiru Gratuito
Eunice Anne Sta. Ana

Department of Chemical Engineering

University of Negros Occidental - Recoletos
Bacolod City

ABSTRACT

The fermentation of pineapple wine is a process that harnesses microbial activity

to convert sugars present in pineapple juice into alcohol. This traditional method of
fermentation has been practiced for centuries, resulting in the production of a unique and
flavorful alcoholic beverage. Pineapple wine fermentation involves the careful selection
and preparation of raw materials, including pineapple fruit, sugar, and yeast, as well as
the maintenance of suitable environmental conditions to facilitate microbial growth and
activity. Through controlled fermentation, yeast metabolizes sugars, producing ethanol
and carbon dioxide as byproducts. The fermentation process not only yields an alcoholic
beverage but also influences its aroma, flavor, and overall quality. Understanding the
intricacies of pineapple wine fermentation is essential for optimizing the production
process and achieving desired sensory characteristics. This study aims to explore the
fermentation process of pineapple wine, focusing on its scientific principles,
methodologies, and outcomes.

Keywords: Sugar, Yeast, Fermentation, Pineapple, Anaerobic respiration, Wine

making, Pineapple wine
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TABLE OF CONTENTS
TITLE PAGE (i)
ABSTRACT (i)
TABLE OF CONTENTS 2
LIST OF FIGURES 3
INTRODUCTION 4
Objectives 5
LITERATURE REVIEW AND THEORY 5
APPARATUS AND EXPERIMENTAL PROCEDURE 9
Reagents 9
Materials 9
Procedural Workflow 9
Procedure 10
RESULTS AND DISCUSSIONS 11
REFERENCES 12
APPENDICES 13
Photo Documentations 13
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LIST OF FIGURES

Figure 1. Production Process for Pineapple Wine Fermentation in Industrial Setting 8

Figure 2. Procedural Workflow 9
Figure 3. Weighing of Pineapple 15
Figure 4. Weighing of Sugar 15
Figure 5. Blending of Pineapple 15
Figure 6. Mixing of Raw Materials 15
Figure 7. Transferring to Fermentation Bottle 16
Figure 8. Fermentation Set-up 16
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INTRODUCTION

The process of fermentation is a crucial aspect of food and beverage production,

serving as a transformative force in the creation of various consumables. This laboratory
report explores the fermentation of pineapple wine, delving into its biochemical
mechanisms, environmental factors affecting fermentation, and the qualitative and
quantitative analysis of its end products. Through meticulous experimentation and
analysis, this report aims to elucidate the intricate processes involved in pineapple wine
fermentation, shedding light on its potential as a flavorful and economically viable
beverage.

Fermentation stands as one of the oldest and most fundamental processes in

human civilization, dating back thousands of years to the dawn of agricultural practices.
From the transformation of grapes into wine to the conversion of grains into beer,
fermentation plays a central role in the creation of numerous alcoholic beverages
(Amerine et al., 2019). Among the myriad fermentable substrates, fruits have long been
cherished for their rich flavors and aromatic profiles, offering a diverse array of
possibilities for fermentation.

Pineapple (Ananas comosus) is a tropical fruit renowned for its sweet and tangy
taste, making it a popular choice for culinary endeavors across the globe (Joshi & Attri,
2018). Beyond its culinary applications, pineapple also serves as a promising substrate
for fermentation, yielding a delightful beverage known as pineapple wine. Pineapple wine,
with its refreshing taste and subtle tropical notes, has gained popularity among
enthusiasts of fermented beverages, offering a unique alternative to traditional wines and
spirits (Fleet, 2020).

The fermentation of pineapple juice into wine primarily relies on the metabolic
activities of various microorganisms, predominantly yeast species, which convert sugars
present in the juice into ethanol and carbon dioxide through anaerobic respiration (Gao et
al., 2024). This transformation involves a series of enzymatic reactions orchestrated by
specific yeast strains, alongside interactions with other microorganisms present in the
fermentation milieu. Additionally, the presence of secondary metabolites and aroma
compounds contributes to the sensory characteristics and overall quality of the resulting
wine (Helmenstine, 2020). In the case of pineapple wine, the sugars present in the fruit
serve as the fermentable substrate, providing the necessary energy for yeast metabolism
(Lim, 2019). As yeast cells metabolize sugars, they produce ethanol and carbon dioxide
as metabolic byproducts, leading to the characteristic alcoholic content and effervescence
of fermented beverages (Ghosh & Yoon, 2017).
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Understanding the intricacies of pineapple wine fermentation requires a

comprehensive examination of various factors influencing the process. Environmental
conditions such as temperature, pH, and oxygen availability play pivotal roles in shaping
the kinetics and outcomes of fermentation (Guerreiro & Cancela, 2017). Additionally, the
choice of yeast strain and inoculation methods can significantly impact the flavor profile
and quality of the final product (Middelberg, 2019) .

This laboratory report delves into the fermentation of pineapple wine, elucidating
the biochemical transformations and physical attributes associated with this process. The
experiment specifically aims to produce pineapple wine using pineapple puree as the
substrate for fermentation. This report seeks to contribute to the body of knowledge in
food science and technology, offering insights into the development of novel fermented
products.

Objectives
This experiment aims to produce wine using pineapple through the fermentation
process of yeast and sugar as the substrate. This report seeks to discuss the biochemical
process involved in the conversion of sugar into alcohol and carbon dioxide as a result of
microbial activity.

LITERATURE REVIEW AND THEORY

Fermentation is a form of biotechnology where beneficial microorganisms are

employed to create valuable commercial products. This process naturally occurs when
sugars in substances like fruits, berries, honey, or palm sap are left exposed to airborne
yeasts in warm environments, leading to the conversion of sugar into alcohol and carbon
dioxide. Controlled fermentation is utilized in the production of wines and beers.
Alcoholic beverages have been a part of various cultures for centuries, playing significant
roles in both personal and social ceremonies. Traditional events such as child naming,
marriage celebrations, and funerals often incorporate alcoholic drinks. Across Africa, a
variety of ingredients including maize, millet, bananas, honey, palm sap, and various
fruits are utilized to produce nutrient-rich beers and wines, with notable examples like
kaffir beer and palm wines (Dittmer & Griffin, 1999).

It's likely that the earliest consumed fermented foods were fruits. While
hunter-gatherers typically ate fresh fruits, during times of scarcity, they likely resorted to
consuming rotten or fermented fruits. This repeated consumption could have led to a
preference for fermented fruits. Historical records suggest that fermented drinks were
produced over 7000 years ago in Babylon, 5000 years ago in Egypt, 4000 years ago in
Mexico, and 3500 years ago in Sudan. There's even evidence of fermented meat products
being made for King Nebuchadnezzar of Babylon (Steinkraus, 2016). China is believed to
be the origin of fermented vegetables and the use of molds like Aspergillus and Rhizopus
in food production. The ancient Chinese book "Shu-Ching," written during the Chou
dynasty (1121–256 BC), mentions the use of "chu," a fermented grain product (Voidarou
et al., 2020).
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Yeasts hold significant economic value, particularly various strains of S. Cerevisiae,

which have been extensively utilized in the production of alcoholic beverages, solvents,
and other chemicals. Yeast, a single-celled fungus or plant-like microorganism, is
ubiquitous in the natural environment, found in water, soil, plants, and air. Belonging to
the group of microbial eukaryotes associated with Ascomycetes, yeasts are notably rich in
protein and Vitamin B. These organisms, as living entities, primarily require sugars,
water, and warmth to sustain themselves, with additional nitrogenous material or albumen
essential for optimal growth. While numerous yeast species exist in nature, S. cerevisiae
is most commonly utilized for baking, as its scientific name suggests its ability to ferment
sugars in grains to produce alcohol and carbon dioxide (Dunn et al., 2005).

Yeasts, crucial in alcoholic fermentation processes, are widespread globally, with

over 8,000 classified strains of this vegetative microorganism. Approximately 9–10
distinct strains, including subclasses, are employed for fermenting grain mashes, all
falling under the type S. cerevisiae. Each strain possesses unique characteristics,
contributing distinct properties to the resulting distillate during fermentation. Within the
S. cerevisiae classification, a select few strains are utilized for fermenting wines, which
are then distilled into brandy. These same strains are also employed in rum production
from sugarcane extracts and beer brewing. Optimal yeast activity occurs in a slightly
acidic environment; thus, the acidity of the mash, juice, sap, or extract designated for
fermentation must be verified. If acidity is inadequate, acid or acid-containing substances
are introduced. For distilled spirits, fermentation typically occurs at temperatures ranging
from 24°C to 29°C over a period of 48 to 96 hours, after which the mash or must is ready
for distillation. The alcohol content of the fermented must typically ranges from 7% to
9% (He et al., 2024).

Fruit juices undergo fermentation to produce wine, an alcoholic beverage. Grapes are
typically favored due to their natural chemical balance, facilitating fermentation without
the need for additional sugars, acids, enzymes, or nutrients. However, various other fruits
such as banana, cucumber, and pineapple are also utilized in wine production. Homemade
wine is crafted from a variety of fruits including apple, pear, strawberry, cherries, plum,
banana, pineapple, oranges, cucumber, watermelon, and guava. S. cerevisiae species are
commonly employed to convert sugar in fruit juices into alcohol and organic acids,
leading to the formation of aldehydes, esters, and other compounds that aid in wine
preservation. Additionally, yeasts sourced from sources like palm wine have been utilized
in fruit wine production (Isitua et al., 2010).

Wine making encompasses three primary phases: pre-fermentation, fermentation, and

post-fermentation operations. For grape wines, pre-fermentation involves crushing the
fruit to extract juice. In white wine production, the juice is separated from the skins,
whereas in red wine, the skins remain with the juice. Juice clarification for white wine
typically involves sedimentation or centrifugation. Yeast is then introduced to the
clarified juice to initiate fermentation. In red wine production, the grape pulp, skins, and
seeds are retained during fermentation to extract color and flavor. Yeast is added to the
mashed pulp (must) during this process (Ayogu, 1999).
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A study done by Reddy and Obulam (2011) investigated the fermentation of mango
juice in a laboratory setting, utilizing a controlled inoculation method with a specific
yeast strain (Saccharomyces cerevisiae 101). The impact of various fermentation
conditions, including temperature, pH, SO2 concentration, and aeration, on yeast growth,
fermentation duration, rate, and volatile compound composition, was examined. Gas
chromatography was employed to analyze the major volatile compounds under different
fermentation conditions.

Temperature was found to significantly influence yeast growth and volatile

compound levels, with higher temperatures resulting in decreased concentrations of ethyl
acetate and certain higher alcohols. The addition of SO2 stimulated yeast growth up to a
certain level but inhibited metabolism at higher concentrations. Ethanol concentration
was observed to increase slightly with the addition of 100 ppm SO2 but decreased with
higher SO2 concentrations. Aeration through shaking increased viable cell count but
reduced ethanol productivity.

Based on the findings, it was concluded that optimal conditions for mango wine
production were a fermentation temperature of 25°C, pH of 5, SO2 concentration of 100
ppm, and initial oxygen in the must. These results align with traditional preferences for
lower alcoholic fermentation temperatures in winemaking.

Berenguer et al. (2016) assessed the three commercial strains of Saccharomyces

cerevisiae yeast (Viniferm Revelación, Viniferm SV, and Viniferm PDM) for producing
pomegranate wine from a blend of Mollar and Wonderfull varieties. The fermentation
process was followed by spontaneous malolactic fermentation. Similar fermentation
trends were observed across all strains regarding pH, acidity, density, sugar consumption,
and the production of ethanol and glycerol. Glucose was fully utilized, leaving fructose
residues. The resulting wine had a high ethanol concentration (10.91 ± 0.27% v/v) and
1.49 g/L of glycerol. Citric acid levels rose by 31.7%, malic acid was depleted due to
malolactic fermentation, and lactic acid levels ranged from 0.40 to 0.96 g/L. Analysis
showed that Viniferm PDM yeast led to less degradation of monomeric anthocyanins
during winemaking compared to the other strains, retaining 34.5% of the total
anthocyanin content from the pomegranate juice blend.

A study done by Ou (2004) explored the nutrient utilization of Saccharomyces

cerevisiae from palm wine using tropical fruits as substrates. Starter cultures were
prepared by growing yeast in progressively larger bottles with pasteurized fruit must.
Microvinification, substrate utilization, and yeast activity assays were conducted. The
soluble solute (SS) content of the juices varied from 10 to 18 Brix, with pineapple must
having the highest SS content (18 Brix) and pawpaw having the lowest (10 Brix). Wine
alcohol levels ranged from 10.6% to 12.6%, while volatile activity ranged from 0.25 to
0.32, and crude protein values ranged from 0.58% to 0.68%. Palm wine yeast and other
strains fermented and utilized fruit must for growth, with specific growth rates from 0.18
to 0.22. Sugar loss in Brix decreased gradually from 20.0-24.0 Brix to 4.8-6.0 Brix.
Pineapple was favored for tropical winemaking, while mango, cashew, and pawpaw were
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suitable for commercial fermentation, albeit requiring more sugar for cashew and
pawpaw than mango juice. Palm wine yeast (OW-11) performed comparably to other
wine yeasts (CBS 8066 and ATCC 4126) in nutrient utilization and growth, suggesting its
suitability for industrial wine production due to its high adaptability.

Application of fermentation in industrial scale encompasses an array of various

processes and technologies to ensure the quality and efficiency of the product. Studies
have shown that elements such as yeast concentration, solubility of solids, and length of
fermentation that occurred influence the quality attributes of pineapple fruit wine
including pH, acidity, alcohol content, and sensory qualities. Furthermore, optimizing the
production process of pineapple wine is observed through effective management practices
and quality control measures to enhance the quality and economic feasibility of the
product. The fermentation process is a vital component of bio processing, employing
enzymes to prompt specific alterations in organic materials. It involves vapors emitted
during the fermentation to condensed in stages in order to enhance the quality of the
products. In large scale production, meticulous measurements and selection of materials
are necessary for quality assurance to maintain consistency and ensure accuracy of the
product throughout the manufacturing process.

Figure 1. Production Process for Pineapple Wine Fermentation in Industrial Setting

During the production process, pineapples are mechanically peeled using automated
apparatus, and crushed for juice extraction. Fermentation tanks are utilized after the
preparation of the raw materials, and the duration set for the process is 15 days. Yeast is
added to fermentation tanks to induce fermentation by converting sugars into alcohol.
Throughout the process, quality assurance is implemented to guarantee the desired quality
of the product.
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APPARATUS AND EXPERIMENTAL PROCEDURE

Reagents
• White sugar
• Wine yeast
• Distilled Water
• Purified Water
Materials
• Caserole
• 1000 mL beaker
• 100 mL beaker
• 500 mL beaker
• Weighing Scale
• Wood ladle
• Measuring spoon
• Fermentation Bottle
• Balloon
• Hot Plate

Procedural Workflow

Preparation of Raw Materials Sterilization of Fermentation

Bottle

Fermentation Process Dissolution of Raw Materials

Storage

Figure 2. Procedural Workflow

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Procedure
1. Preparation of Raw Materials
In the preparation of raw materials, 50 mL of water was measured and 5 g of wine
yeast was added. The yeast solution was allowed to rest to proof or until it doubled in
amount. Additionally, 500 g of pineapple chunks and 250 g of sugar were weighed, and 1
teaspoon of calamansi juice was measured for the purpose of introducing an additional
flavor to the wine. The pineapple chunks were then blended in a blender to produce
pineapple puree, which served as the substrate for the fermentation process.

2. Sterilization of Fermentation Bottle

In the sterilization of the container, 1 L of distilled water was boiled in a beaker.
The fermentation bottle was washed thoroughly to remove any dirt or debris present.
Subsequently, the boiling water was poured into the fermentation bottle, where it was left
to rest until the solution intended for fermentation was prepared and ready to be
transferred.

3. Dissolution of the Raw Materials

The dissolution phase involved heating 1 L of purified water until it reached the
boiling point, after which the sugar, pineapple puree, and calamansi juice were added.
The heat was turned off, and the solution was stirred until the sugar was completely
dissolved. Following this, the solution was allowed to cool down to room temperature.

4. Fermentation Process
For the fermentation process, 4 teaspoons of proofed yeast were added to the
solution and thoroughly stirred. The solution was then transferred into the sterilized
fermentation bottle, and a balloon was securely placed over the mouth of the bottle. The
solution was left to ferment for a duration of 2-3 weeks.

5. Storage
The pineapple wine obtained after fermentation was transferred in the container
for storage.
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RESULTS AND DISCUSSIONS

In the fermentation process, the strain of yeast utilized is Saccharomyces

cerevisiae, which is generally the yeast employed in wine making. During fermentation,
yeast generates CO2 as a natural byproduct of converting sugars into alcohol. Following
a week of fermentation, the balloon affixed to the bottle exhibited minimal inflation,
indicating that CO2 production was relatively low. Nonetheless, it can be inferred that the
yeast successfully metabolized the sugar, yielding alcohol and CO2. The alcohol content
in the fermented product remained undetermined, as no standardized procedure for
measurement was conducted. The observation on minimal balloon inflation is caused by
several factors contributing to this result and its implications for the overall fermentation
process.

Firstly, the limited production of carbon dioxide suggests that the microbial
activity responsible for fermentation may have been deficient and several factors could
have influenced this including temperature, nutrient availability, and contamination.
There is a possibility that the temperature within the fermentation environment was not
within the ideal range for the microbial strains involved since microorganisms are highly
sensitive to temperature fluctuations and deviations from the optimal range can
significantly affect their metabolic activity. Additionally, insufficient nutrient availability
or imbalance sugar to yeast ratio within the solution may have hindered microbial growth
and fermentation efficiency. Carbon dioxide production serves as an indicator of ongoing
fermentation activity and the conversion of substrates into desired end products. The
subdued release of carbon dioxide suggests that fermentation may have been incomplete.
Inadequate fermentation time or premature termination of the experiment could have
impeded the attainment of desired results. Moreover, the balloon attached to the bottle
was not secured properly which may have caused some CO2 gas to escape and factors
such as oxygen exposure or contamination by unwanted microorganisms may have
compromised the fermentation process, leading to little amount of carbon dioxide
produced.

Depending on the desired end products, the extent of carbon dioxide production
may directly impact the quality, quantity, and properties of the final fermented product. In
industries such as food and beverage production, where fermentation is employed for the
synthesis of various products, optimizing fermentation conditions to enhance carbon
dioxide production is paramount for achieving desired product characteristics and market
competitiveness.

Overall, the limited release of carbon dioxide observed in the fermentation

experiment underscores the complex interplay of various factors influencing microbial
activity and fermentation efficiency. By critically analyzing these results and exploring
potential contributing factors, valuable insights can be gained into the dynamics of
fermentation processes and advance the understanding of optimizing fermentation
conditions for improved outcomes.
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CONCLUSION

In conclusion, the fermentation experiment of pineapple wine resulted to unexpected

results characterized by minimal carbon dioxide production. Despite initial expectations
and efforts to stimulate fermentation activity, the process exhibited a minimal release of
carbon dioxide gas. This outcome may have been influenced from various factors,
including the choice of yeast strain, fermentation conditions, and the composition of the
pineapple juice substrate. While the production of minimal carbon dioxide may seem
advantageous in some contexts, such as reducing environmental impact or preserving
certain flavor profiles, it nonetheless signifies a deviation from typical fermentation
patterns and raises questions about the efficiency and completeness of the fermentation
process. Further investigation is warranted to elucidate the underlying mechanisms
behind this phenomenon and to determine its implications for the final product's sensory
attributes, alcohol content, and overall quality. By unraveling the complexities of this
unexpected outcome, future research endeavors can enhance our understanding of
fermentation dynamics and pave the way for more controlled and predictable outcomes in
pineapple wine production.
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Ayogu, T. (1999). Evaluation of the performance of a yeast isolate from Nigerian palm
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Berenguer, M., Vegara, S., Barrajón, E., Saura, D., Valero, M., & Martí, N. (2016).
Physicochemical characterization of pomegranate wines fermented with three
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Gao, J., Liu, J., & Pang, X. (2024). Characterization of the color, physicochemical
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Helmenstine, A. M., PhD. (2020, October 2). What is fermentation? Definition and
examples. ThoughtCo. https://www.thoughtco.com/what-is-fermentation-608199
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Isitua, C, C., Ibeh, & G, M. (2010). Novel method of wine production from banana (Musa
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APPENDICES

Photo Documentations

Figure 3. Weighing of Pineapple Figure 4. Weighing of Sugar

Figure 5. Blending of Pineapple Figure 6. Mixing of Raw Materials

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Figure 7. Transferring to Fermentation Bottle Figure 8. Fermentation Set-up