BIO Web of Conferences 155, 01006 (2025) https://doi.org/10.
1051/bioconf/202515501006
10th ICCC 2024
Prebiotic ability of musa balbisiana colla stems
on the growth of Pediococcus acidilactici in vitro
and probiotic encapsulation
Ni Made Ayu Suardani Singapurwa1*, I Putu Candra1, A.A. Made Semariyani1, Ni Wayan
Nursini2, Purwaningtyas Kusumaningsih2, Gusti Ngurah Oka Jiwantara1 and Sang Ayu
Made Agung Prasetiawati Djelantik1
1 Food Technology and Agricultural Products Department, Warmadewa University, Denpasar, Bali
2 Nutritional Science Department, Dhyana Pura University, Badung, Bali
Abstract. Musa balbisiana Colla banana is a plant that has many benefits,
one of which is useful as a traditional medicine using its stems. Banana stems
have potential as a source of prebiotics because they contain carbohydrates
and oligosaccharides which can support the growth of good bacteria in the
digestive tract. Probiotic encapsulation was carried out to determine the role
of Musa paradisiaca stems as a prebiotic in digestion. The encapsulation
process is carried out using chitosan, carrageenan, and maltodextrin. The
results showed that the prebiotic banana stem Musa paradisiaca could
increase the growth of Lactobacillus sp. significantly at certain
concentrations, with the most optimal results at a concentration of 6.89%
compared to the control. Based on test results, Musa paradisiaca stem flour
contains a fructan percentage of 0.14 ± 0.001%. The presence of fructan
compounds makes Musa paradisiaca stem flour function as an effective
source of natural prebiotics. The test results showed that the biocapsulation
contained 2.7 x 103 cfu/gram of lactic acid bacteria. This shows that the
bioencapsulation process of lactic acid bacteria was successful.
Encapsulation can protect probiotics from extreme conditions during
processing and storage, as well as from acidic pH in the digestive tract.
1 Introduction
Balinese people have various ways of processing vegetables to meet their needs, which are
grouped as vegetables without soup, vegetables with little soup, and vegetables with a lot of
soup. Banana stem vegetable and roroban vegetable are two types of Balinese cuisine that
have a lot of soup made from young banana tree trunks. Banana trunk is another name for
banana tree saplings or banana trees with young trunks in Bali.
For people outside Bali, they may not know that banana stems or young banana trees can
be processed as side dishes. Banana stems are a favorite food of the Balinese people which
are cooked when there is a big celebration or ceremony involving many members of the
* Corresponding author: a.suardani@gmail.com
© The Authors, published by EDP Sciences. This is an open access article distributed under the terms of the Creative Commons
Attribution License 4.0 (https://creativecommons.org/licenses/by/4.0/).
BIO Web of Conferences 155, 01006 (2025) https://doi.org/10.1051/bioconf/202515501006
10th ICCC 2024
community. So, it is cooked in large portions, with complete spices (basa genep) and lots of
gravy, so that it can meet the needs of many people [1]
Banana is a plant that has many benefits, one of which is useful as a traditional medicine
by using its stem. Ambon Banana Stem is used as a natural ingredient to reduce the use of
synthetic drugs. The stem has active compounds, namely flavonoids and tannins, which play
a role in preventing the side effects of indomethacin. Banana stem flavonoids can function
as antioxidants and anti-inflammatories. Flavonoids work by slowing down the inflammation
process through an inhibitory effect on the arachidonic acid metabolism pathway and the
formation of prostaglandins. While tannins play a role in stopping bleeding [2].
Prebiotics are food ingredients that cannot be digested in the small intestine and when
they reach the large intestine will be fermented by colon bacteria into short chain fatty acids
(SCFA) such as acetate, lactate, butyrate, propionate, and also gases (such as CO 2, methane,
and hydrogen). These short chain fatty acids will be absorbed and metabolized by the body
[3]. Prebiotics are generally in the form of dietary fiber [4]. In addition, the most potential
prebiotics also consist of carbohydrates. Food ingredients that are prebiotics can be
vegetables, tubers, and fruits [5]. Several previous studies on prebiotic sources or products
have used many high dietary fiber and carbohydrate ingredients. One of them is from fruit,
namely banana flour, which in previous studies has been shown to have benefits for the
growth of the probiotic Lactobacillus casei. There is also other research on the use of
prebiotic extracts from Rumbia fruit (Metroxylon sago Rottb), the results of which show the
influence of prebiotic sources on ration consumption, final weight gain, and other positive
effects on broiler chickens [6]
So far there has been no in-depth information or research on the potential of banana stems
as a source of prebiotics. Prebiotic tests can use LAB (Lactic Acid Bacteria), especially from
the genus Pediococcus which is a normal flora. One of them used in this study is Pediococcus
acidilactici which is a bacteria with the ability to inhibit contamination from pathogenic
microorganisms and toxin producers because it can produce lactic acid and lower pH. And
has the ability to produce bacteriocins that function as antibiotics [7]. Based on the
explanation above, this study was conducted with the aim of determining the ability of banana
stem prebiotic sources based on their prebiotic tests on the growth of Pediococcus acidilactici
in vitro. The results of this study are expected to provide additional information on the
benefits of banana stems and increase the use of natural ingredients, one of which is banana
stems as a source of prebiotics.
2. Research methods
2.1 Research Location
This research was conducted at the Laboratory of the Faculty of Agriculture, Warmadewa
University, the Laboratory of Dhyana Pura University and the Analytical Laboratory of
Udayana University. Banana stems were obtained from Antap Village, Selemadeg District,
Tabanan.
2.2 Tools and materials
The materials used in this study include: banana stem fruit, Inulin Powder, cold water (0℃),
MRS agar media (de Man, Rogosa, Sharpe), MRS broth media, distilled water, 0.85% NaCl,
and Pediococcus acidilactici bacterial isolates. The tools used in this study include: petri
dishes, test tube racks, test tubes, LAF (Laminar Air Flow), tubes, spectrophotometers,
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BIO Web of Conferences 155, 01006 (2025) https://doi.org/10.1051/bioconf/202515501006
10th ICCC 2024
micropipettes, beakers, blenders, knives, 80 mesh sieves, Erlenmeyer flasks, ovens,
autoclaves, vortexes, bunsen, ose needles, analytical scales, tips, stirrers, hotplates, colony
counters, measuring cups, spatulas, aluminum foil, rubber, paper, plastic, cotton, and trays.
2.3 Research Procedures
Research Stages of Making Banana Stem Flour are prepared, namely selected from the unripe
Musa balbisiana Colla banana variety. Furthermore, washing, peeling, slicing approximately
22 mm, and weighing as much as 500 grams. Then given a soaking treatment in cold water
0 ℃ for 10 minutes, then drying in an oven for approximately 3 days at a temperature of 45
℃-50 ℃. After drying, flour is made using a blender and sieved using an 80 mesh sieve.
Culture Rejuvenation Bacterial culture rejuvenation begins with the preparation of a growth
medium, namely MRS broth media by weighing 2.6 grams of media powder and dissolving
it in 50 ml of distilled water. Furthermore, the media is homogenized with a magnetic stirrer,
then 5 mL of media is poured into a test tube and sterilized with an autoclave at a temperature
of 121 ° C for 15 minutes. Furthermore, the media is removed and cooled. The frozen
bacterial culture stock was taken as much as 1 loop and inoculated into 5 mL of MRS broth
media then incubated for 24-48 hours at 37°C. Growth was indicated by turbidity in the media
[8].
Carrageenan 1%, maltodextrin 1.5% and flour 1.5% were dissolved in 30 mL of distilled
water (Solution A), then heated at 96°C for 5-6 minutes. The solution was cooled to 45°C
while stirring. Making a 2% chitosan solution is by weighing 2 grams of chitosan and
dissolving it with 1% acetic acid (Solution B). Furthermore, the addition of Pediococcus
acidilactici bacterial cell suspension (1:6) to the mixture of solution A and solution B. The
mixture of ingredients was stirred with a magnetic stirrer for 2 hours and then ovened for 24
hours at 40°C until the banana stem flour bioencapsulation powder was produced.
2.3 Prebiotic Characteristics Test
The prebiotic characteristic test in this study was carried out by observing the amount of
Pediococcus acidilactici bacterial growth in banana stem flour bioencapsulation powder. The
test began with the preparation of MRS broth media, weighing 15.6 grams of media and
dissolving it in 300 mL of distilled water and homogenizing it. The banana stem flour
bioencapsulation powder was inoculated into 10 mL of MRS broth media that had been added
with banana stem flour bioencapsulation powder. Furthermore, it was incubated at a
temperature of 37ºC for 24-48 hours. The population of Pediococcus acidilactici in banana
stem flour was calculated using the spread method [8]. The bacteria that grew were then gram
stained to ensure that the growth of the bacteria was lactic acid bacteria.
3. Results and Discussion
Musa paradisiaca stem flour has the ability as a prebiotic because of its carbohydrate and
oligosaccharide content that can support the growth of good bacteria in the digestive tract.
The yield of banana flour bioencapsulation was 0.5 ± 0.02%. The low yields produced are
caused by several factors. The inoculum concentration used in the bioencapsulation process
has a significant effect on the results. Using optimal concentrations can increase the number
of microorganisms involved thereby increasing the production of the desired metabolite.
Factors such as pH, temperature and fermentation time also greatly determine the results.
Lactic acid bacteria grow well at pH 5-6, where they can produce effective lactic acid and
bacteriocins. A decrease in pH below 4.5 can kill pathogenic bacteria, while a pH above 6.5
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BIO Web of Conferences 155, 01006 (2025) https://doi.org/10.1051/bioconf/202515501006
10th ICCC 2024
can inhibit the growth of Lactic acid bacteria itself [9]. At a temperature of 37°C, Lactic acid
bacteria shows optimal growth. On the other hand, at temperatures below 10°C or above
45°C, the growth of these bacteria is inhibited or does not occur at all. Probiotic encapsulation
aims to protect Lactic acid bacteria from unfavorable environmental conditions, including
temperatures [10]. Encapsulation aims to protect bacteria from high temperatures that can
kill bacterial cells or damage their cellular structure. The substrate used for bioencapsulation
must be rich in nutrients and suitable for microbial growth. Substrate quality can influence
the interaction between microorganisms and their binding materials, as well as the
availability of nutrients necessary for optimal growth [11]. In this stem research, the substrate
used was banana flour.
Based on the test results, Musa paradisiaca stem flour contains a percentage of fructan of
0.14 ± 0.001%. Fructans can be found in various food sources, including bananas, which
contain inulin and FOS [12]. The presence of fructan compounds makes Musa paradisiaca
stem flour function as an effective source of natural prebiotics [13]. The requirements for
good prebiotics are that they cannot be hydrolyzed in the upper gastrointestinal tract, can be
digested by good bacteria in the colon, so that they can suppress the growth of pathogenic
bacteria. Lactic acid bacteria have the enzyme alpha-galactosidase which can break down
alpha-galactosidase bonds which can break down polysaccharides into galactose, glucose,
and fructose [12].
Probiotic encapsulation was conducted to determine the role of Musa paradisiaca stems
as prebiotics in digestion. The encapsulation process was carried out using chitosan,
carrageenan, and maltodextrin. The encapsulation results were tested for the presence of
lactic acid bacteria using the spread method. The test results showed that the biocapsulation
contained 2.7 x 103 cfu/gram of lactic acid bacteria. This indicates that the bioencapsulation
process of lactic acid bacteria was successful. The higher the total growth of lactic acid
bacteria microbes, the better the bioencapsulation process was. Encapsulation can protect
probiotics from extreme conditions during processing and storage, as well as from acidic pH
in the digestive tract. Encapsulation can increase the stability of probiotics during storage and
distribution of products. The encapsulating material used is chitosan which can form a matrix
that protects probiotic cells from damage [14], [15]. By using encapsulation techniques,
probiotics can be added to various food products without changing the taste or texture [16],
[17]. This will make it easier to consume probiotics in more diverse forms, thus increasing
consumer compliance with probiotic consumption [18]. Encapsulation protects probiotic
cells from detrimental factors such as stomach acid, emping salts, and high temperatures that
can reduce their viability. Research shows that encapsulated Lactobacillus plantarum and
Streptococcus thermophilus bacteria have better resistance to stomach acid conditions
compared to non-encapsulated ones, with a lower decrease in viability [19]. The
encapsulation process creates a protective layer around the probiotic cells, thereby helping to
maintain their integrity and stability during storage and processing [7]. This is especially
important in food products that undergo heating or storage for long periods. Encapsulation
using materials such as chitosan and alginate has been proven to be able to maintain the
number of bacterial colonies in simulated gastric acid fluid [20]. Encapsulation allows control
of the release of probiotics in the digestive tract. By using appropriate encapsulant materials,
probiotics can be released gradually according to pH conditions in the gastrointestinal tract,
ensuring that more probiotic cells reach the large intestine, where they can provide health
benefits [21]. The results of probiotic encapsulation were continued with gram staining to
ensure that lactic acid bacteria were well encapsulated. Lactic acid bacteria are gram-positive
bacteria in the form of bacilli (rods) which can be seen in Figure 4.
4
BIO Web of Conferences 155, 01006 (2025) https://doi.org/10.1051/bioconf/202515501006
10th ICCC 2024
Fig 1. Probiotic Encapsulation Fig 2. Growth of lactic acid bacteria from
probiotic encapsulation
Fig 3. Total Lactic Acid Bacteria Testing using the Fig 4. Results of gram stain test of lactic acid
TPC method bacteria in probiotic encapsulation.
4. Conclusion
The results showed that the prebiotic banana stem Musa paradisiaca could increase the
growth of Lactobacillus sp. significantly at certain concentrations, with the most optimal
results at a concentration of 6.89% compared to the control. Based on test results, Musa
paradisiaca stem flour contains a fructan percentage of 0.14 ± 0.001%. The presence of
fructan compounds makes Musa paradisiaca stem flour function as an effective source of
natural prebiotics. The test results showed that the biocapsulation contained 2.7 x 103
cfu/gram of lactic acid bacteria. This shows that the bioencapsulation process of lactic acid
bacteria was successful.
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