Accepted Article

DR. EMILIA JANISZEWSKA-TURAK (Orcid ID : 0000-0002-5653-0436)

DR. KATARZYNA POBIEGA (Orcid ID : 0000-0003-3532-8666)

Article type : Original Manuscript

The influence of Lactobacillus bacteria type and kind of carrier on the properties of spray-dried
microencapsules of fermented beetroot powders
Running title: Encapsulated fermented beetroot powder
Emilia Janiszewska-Turak1, Łucja Hornowska1, Katarzyna Pobiega2, Małgorzata Gniewosz2,
Dorota Witrowa-Rajchert1
1 Department of Food Engineering and Process Management, Institute of Food Sciences, Warsaw
University of Life Sciences (SGGW), Warsaw, Nowoursynowska 159c, 02-776 Warsaw, Poland
2 Department of Food Biotechnology and Microbiology, Institute of Food Sciences, Warsaw
University of Life Sciences (SGGW), Warsaw, Nowoursynowska 159c, 02-776 Warsaw, Poland

Emilia Janiszewska-Turak - emilia_janiszewska_turak@sggw.edu.pl Orcid: 0000-0002-5653-0436

Katarzyna Pobiega – katarzyna_pobiega@sggw.edu.pl Orcid: 0000-0003-3532-8666
Małgorzata Gniewosz – malgorzata_gniewosz@sggw.edu.pl Orcid: 0000-0003-2282-8231
Dorota Witrowa-Rajchert – dorota_witrowa_rajchert@sggw.edu.pl Orcid: 0000-0002-0937-3204
*Corresponding author:
Ph. D. eng. Emilia Janiszewska-Turak, tel.: +48 22 5937566, fax.: +48 22 5937576

The peer review history for this article is available at https://publons.com/publon/10.1111/ijfs.14915

This article has been accepted for publication and undergone full peer review but has not been
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differences between this version and the Version of Record. Please cite this article as doi:
10.1111/ijfs.14915
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 Abstract
Accepted Article
The aim of this study was to evaluate the influence of bacteria type and carrier addition during
microencapsulation process by spray drying on the microorganism survival and physicochemical
properties of fermented beetroot juice powder. Selected bacteria were: L.fermentum, L.plantarum
and L.brevis and its mixture in proportion 1:1:1. Fermented beetroot juice containing selected
LAB bacteria was spray-dried at 160 o C. Maltodextrin and gum Arabic were used as drying
carriers at 10% (w/w). All powders were stable (low water activity bellow < 0.25, high dry matter
content 95-98%). Bacteria type had the main influence on the chemical properties and amount of
bacteria. The highest bacteria content was observed in powders with L.brevis as a starter, the same
was observed for polyphenols and betalain content. However, the method of drying fermented
juices still needs to be refined due to the 50% reduction in the number of microorganisms during
the drying process.

Key words: Lactobacillus, betanin, colour, dedicated fermentation, microencapsulation, spray-

drying

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 1. Introduction
Accepted Article In recent years, because of an increase in number of people who declare intolerance to
lactose and who have allergies to milk components, an increase of interest in fermented vegetables
and fruits products is observed (Alves et al,. 2016). Lactic acid bacteria (LAB) are responsible for
the fermentation. They are rod-shaped or coarse-shaped bacteria, grow in anaerobic conditions,
they are gram positive, and do not produce spores. Their growth depends on many factors
especially the species. If LABs metabolize sugars (usually glucose) into lactic acid and other
products, e.g. acetic acid, ethanol and carbon dioxide, hetero-fermentation is observed (Hurtado et
al,. 2012, Nuraida 2015). After hetero-fermentation process obtained fermented vegetables or
fruits are protected because of presence in food produced during fermentation substances such as
an alcohol, lactic acid, and acetic acid. Fermentation takes place only when the microorganisms
have access to a suitable substrate from which they can derive a source of carbon and favourable
pH conditions (Pérez-Armendáriz and Cardoso-Ugarte 2020).
In the production of juices, there is a spontaneous fermentation taking place due to the
presence of the natural microflora of the raw material or forced fermentation with the use of lactic
acid bacteria, i.e. starter cultures. The appropriate selection of bacteria affects the course of the
fermentation process and determines the final quality of obtained juice (Zielinska and
Waszkiewicz-Robak 2015). Malik et al,. (2019) used L. plantarum, L. acidophilus and L. casei to
produce fermented carrot and beet juice, while Panda et al,. (2017) used L. fermentum bacteria to
produce prickly pear juice. The above-mentioned species are the most frequently chosen for using
to fermentation process.
In Poland, pickling of cabbage, carrots, tomatoes and red beets is very popular. Red beets,
like all vegetables, are rich in minerals, vitamins, fibre and biologically active substances - thanks
to which they have a health-promoting effect and their presence in the diet reduces the incidence
of cardiovascular diseases and reduces the risk of cancer (Kazimierczak et al,. 2014). They are
characterized by a high content of folic acid, iron, sodium, potassium, magnesium and vitamins A,
C, B7, B3 (da Silva Carvalho et al,. 2016). The resulting beetroot juices contain large amounts of
flavonoids, phenolic acids, vitamin C and bioactive compounds, which include: betanin and
vulgaxanthin-I, which together give beetroot its colour. Betanin, belonging to the betacyanin
colorants, is responsible for the red colour, while vulgaxanthin-I is one of the yellow pigments and
belongs to the betaxanthin group. These water-soluble betalains belong to the compounds with
antioxidant activity; they are even able to extinguish reactive oxygen species (Klewicka 2012,

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 Kazimierczak et al,. 2016, Guadarrama-Lezama et al,. 2014, Kazimierczak et al,. 2014). However,
Accepted Article
betalains can decompose under the influence of temperature, oxygen, pH, metal ions or the
presence of light (Sawicki and Wiczkowski 2018). In order to protect the sensitive components of
beetroot juice, a microencapsulation process using spray drying is often used, thanks to which a
powder with a low water content is obtained. A lot of research has been made with spray drying of
non-fermented beetroot juice. In literature could be found testing of different carriers type as well
as spray drying process condition changes (Guadarrama-Lezama et al,. 2014, Janiszewska 2014,
Nemzer et al,. 2011, Bazaria and Kumar 2016a, Bazaria and Kumar 2016b, Bazaria and Kumar
2018, Carmo et al,. 2018).
However, in the juice obtained from fermented beets, additionally there are lactic acid
bacteria presence.In order to use spray drying technology to produce probiotic powder, challenges
such as heat, oxidative, osmotic and drying stress must be overcome, especially with sensitive
probiotic strains. During spray drying, the membrane of probiotic cells is damaged, which causes
them to heat up and dehydrate. Due to the high sensitivity of probiotic bacteria, and hence
numerous damage to cell membranes and exposing them to oxidative stress, it is important to
select process parameters and carriers that will have a significant impact on the viability of
probiotics. In order to limit damage to bacterial cells, a microencapsulation process with a carrier
is used. The most frequently chosen carriers are maltodextrin, gum Arabic or monoglycerides. The
process is based on the retention of substances inside the material to protect against the derivative
effects of environmental factors (Lapsiri et al,. 2012, Kavitake et al,. 2018, Colín-Cruz et al,.
2019). This carriers was used for encapsulation of Blackberry juice with Lactobacillus acidophilus
with maltodextrin and gum Arabic (Colín-Cruz et al,. 2019) or Lactobacillus Acidophilus
(Arepally and Goswami 2019, Arepally et al,. 2020), as well as jussara juice with Bifidobacterium
spp. Lactis with inulin or its mixture with maltodextrin (Paim et al,. 2016). What more, among the
various process variables, the inlet and outlet air temperature is a critical variable for probiotic
encapsulation. Prior to the research work on fermented beetroot juice spray drying, preliminary
tests to reduce the inlet air temperature were carried out. However, using a temperature of 120°C
and a feed speed of 8·10-7 m3/s, satisfactory results were not obtained, the powder removed from
the cyclone was burnt and most of the material was deposited on the walls of the dryer. In further
tests, the temperature at the inlet was increased, and the outlet temperature reduction were made
by increasing the speed of feeding the solution to the disk. Also in this case, no powder was
obtained that could be stored, the obtained powder immediately after opening the container

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 changed state and stuck to the chamber. In view of the above, only the medium used for spray
Accepted Article
drying was changed. The choice of carriers and their proportions was made on the basis of
research on beetroot juices microencapsulation and on the most frequently used carriers for
probiotic bacteria microencapsulation.
Maltodextrin (MD) was selected because of its ability for prevention to thermal stress, its
low cost, and its absorbability and digestibility. One of the most common additives for MD is gum
Arabic (GA), known from its inhibits dehydration of cellular components and protection of
microbial cells during drying and storage. At the same time, gum Arabic significantly increases
the glass transition temperature, reduces hygroscopicity and caking (Zhang et al,. 2020a, Zhang et
al,. 2020b, Arepally et al,. 2020).
That is the reason the aim of the study was to test the effect of used LAB bacterial strains
type and the carrier type addition in microencapsulation by spray drying process on the amount of
bacteria and active substances in the obtained powders. Moreover, the physical properties of the
powders were also investigated, to check the stability of the powders..
2. Materials and Methods
2.1. Materials
Beetroot was purchased from local market in Warsaw. Low-crystallised maltodextrin
DE=11 (MD) (PPS “PEEPES” S.A., Łomża, Poland) and gum Arabic (GA) (Hortimex Sp. z o.o.,
Konin, Poland) were used as the carriers. Reference strains were obtained from the American
Type Culture Collection (ATCC, Manassas, VA, USA) and Collection of Industrial
Microorganisms (KKP, Warsaw, Poland).
2.2. Preparation of inoculums
Three probiotic bacteral strains (LAB) (Lactobacillus brevis KKP 804, Lactobacillus
fermentum KKP 811, Lactobacillus plantarum ATCC 4080) were used in the study. The bacterial
strains were cultured on de Man Rogosa and Sharpe Agar (MRS; Biomaxima, Poland) and
incubated at 37 °C ± 1 °C for 24 h. Bacterial inocula were prepared in sterile 0.85% NaCl (w/v)
solution to reach a population of approximately 1 × 109 CFU/mL. The study was carried by
separately inoculating L. brevis (LB), L. plantarum (LP), and L. fermentum (LF) strains and
combination of tested strains in the ratio of 1: 1: 1 (L MIX).
2.3. Fermentation process
Beetroot balls were washed with water, manually peeled and sliced. Those sliced beetroot
was placed into glass jars and 2% solution of NaCl in a proportion 1:1. The inoculum was added

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 to the prepared beets in the amount of 1% of the water volume, which corresponded to the
Accepted Article
bacterial content of approximately 1 × 107 CFU/mL. This is the minimum concentration of
probiotic bacteria in food that is able to induce regulatory effects (Vinderola and Reinheimer
2000). Jars was closed for obtained anaerobic conditions. After that all experiments was left for 5-
7 days (the pH into jars was tested from 5th to 7th day till achieving pH of minimum 3.9). After
obtaining pH level, fermentation process was stopped by placing jars into the refrigerator for at
least 12 h. After that juice from beetroot was pressed. All experiments were done in duplicate.
After fermentation juice from fermented beetroot was obtained with juicer NS-621CES
(Kuvings, Korea). To juice (about 3oBx) carrier was added. As a carriers, maltodextrin (MD) and
its mixtures with gum Arabic (GA) in proportion MD:GA = 2:1 were used. In these experiments,
10% of carrier material or its mixture was used. Final concentration of solid soluble content in
solutions was set at about 13oBx. All drying experiments were done in duplicate.
All solutions were kept for 24 h before spray drying to for complete hydration of the carrier
and check its stability (Chranioti et al,. 2015)
2.3. Spray drying
Semi-industrial spray drier LAB S1 (Anhydro, Copenhagen, Denmark) equipped with spray
drying disc which diameter was 0.064 m was used for experiments. The following parameters of
the drying process were applied: spray disc speed of 39,000 rpm and raw material flux rate of
8·10-7 m3/s, inlet air temperature 160 º C at a constant air flow of 0.055 m3/s. Outlet air
temperature was at level 70±2 º C. Powder was collected in the cyclone and then transferred to
glass "twist-off" jars. Dried powders were stored in jars in a dark place. All experiments were
done in duplicate.
3. Analytical methods
3.1. Enumeration of lactic acid bacteria LAB
The total count by pour plate method was used for enumerating the viable cells. Fermented
juice samples were serially diluted using sterile saline to achieve 15–300 colonies on MRS agar
plates at 37°C ± 1 °C for 48 h ± 4h of incubation. The juice and dried juice samples (1 g) were
dissolved in 9 mL saline and dilutions were prepared as before. The number was recorded as Log
CFU per g. The samples were analysed in triplicates.

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 3.2. Physicochemical properties of fermented beetroot powders
Accepted Article
3.2.1. Dry matter content and water activity
Dry matter content (DM) in powders was measured by the gravimetrical method for solutions
in a 75oC for 24h, for powders in a 105oC for 4h.
The water activity was measured at a temperature of 25°C by a Rotronic Hygroscop DT
hygrometer (Switzerland). All measurements were done in triplicate.
3.2.2. Morphology and size of microcapsules
The morphology of microcapsules was tested by taking images on a desktop scanning electron
microscope, Hitachi TM3000 (Hitachi High-Technologies Corporation, Tokyo, Japan) operated at
an accelerating voltage of 15 kV.
Size of the particles was measured by the laser diffraction method on a Cilas Particle Size
Analyzer 1190 (Orléans, France). Measurements were made in liquid dispersion created using
ethyl alcohol (in which powders do not dissolved). Each time the obscuration was set at 8-10%.
This test was made in 3 repetitions. The average diameter of particles, d50 (median) was measured.
3.2.3. Colour of powders
Analysis of powders colour was made in CR-5 (Konica Minolta, Japan) in CIE L*a*b*
system. Measurement parameters: iluminant D65, angle 2°, calibration with white colour as was
presented by Janiszewska (2014). All measurements were made in triplicate.
3. 3. Quantification of betalain and polyphenols content
Betalain content

Quantification of betalain was performed by the spectrophotometric method described by

Janiszewska and Włodarczyk (2013) A Helios Gamma spectrophotometer (Thermo Spectronic,
Cambridge, UK) was used in this measurement. Pigments were extracted from the sample with a
phosphate buffer at pH 6.5 by shaking for 10 minutes 0.5g sample (solution or powder) with 50mL
of phosphate buffer.
The determination of betalain concentration, i.e. red and yellow pigments, was calculated
in terms of betanin (mg betanin/100g d.m.) and vulgaxanthin-I (mg vulgaxanthin-I /100g d.m.),
respectively. Pigment content calculations were based on the absorption values A1%, which were
1120 for betanin (at 538 nm) and 750 for vulgaxanthin-I (at 476 nm). According to the
methodology, absorbance at 600 nm was measured and used to correct the amounts of impurities
(Janiszewska and Włodarczyk 2013).

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 Total phenolic content (TPC)
Accepted Article The extracts were prepared according to methodology presented by Bednarska and
Janiszewska-Turak (2020) and Nowacka et al,. (2014). Water extracts of spray-dried powders
were prepared for measurement: For this purpose 0.3 g of material (±0.0001g precision) were
weighted into beaker, and then 30 ml of distilled water was added. The samples were mixed. Then
was infiltrated into volumetric flasks and filled to 50 ml with distilled water. Polyphenol content
was measured spectrophotometricaly according to Folin-Ciocalteau’s method (Van Der Sluis et al,.
2002). The absorbance was measured in spectrophotometer Thermo Spectronic Helios Gamma
(Thermo Fischer Scientific, USA), with wave length 750 nm, in relation to the sample without
extract. The measurements were repeated two times.
3.4 Storage
All powders was kept for 3 months in normal conditions (25oC, humidity from 40-50%,
without light). Dry matter, water activity and amount of bacteria were measured in stored powders.
3.4. Statistical methodology
Data are presented as mean ± standard deviation. Significance of inter-group differences was
determined by one-way analysis of variance (ANOVA) with Statistica 13.3 software. Individual
group differences were identified using the Tukey multiple range test at a significance level of
0.05.
4. Results and discussion
4.1. Microbiological analysis

As shown in Table 1, the probiotic bacteria L. brevis, L. plantarum, and L. fermentum as well as
their mixture can be used for beet fermentation. The squeezed juices obtained after fermentation of
beet 6.30-7.92 log CFU/g. These juices can be classified as probiotic products as the LAB
bacterial content exceeds 6 log CFU/ml (Malik et al,. 2019). The strains of L. brevis and L.
plantarum showed better fermentation properties. After drying the juice using maltodextrin and
maltodextrin with gum Arabic in the ratio of 2:1, a decrease in the number of bacteria by 3.31-4.83
log cycles was observed. During 3 months of storage of the powders, no changes in the number of
lactic acid bacteria were observed, the LAB number was in the range of 2.13-3.80 log CFU/g.
Slightly more bacteria remained in the juice dried using the combination of maltodextrin and gum
Arabic than in the dried juice using only maltodextrin. The method of drying fermented juices still
needs to be refined due to the approximately 50% reduction in the number of microorganisms

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 during the drying process. It could be related to the outlet temperature which was set up at 70 o C,
Accepted Article
what could be one from the factors that affect the survival of bacteria. In addition to the carrier, the
content of lactic acid bacteria in the product may be influenced by the acidity of the juice, as well
as the oxygen concentration or the parameters of spray drying (Shah 2007, Barbosa and Teixeira
2017). Until now, dried beetroot juices have been studied frequently, while dried beet-fermented
juices have not been analyzed. In dried fermented apple juices, increasing maltodextrin to 20%
resulted in LAB survival at 90% (Pereira et al,. 2014). In turn, the dried orange juices with the
highest survival rate was achieved spouted bed drying at low feed flow rates using maltodextrin as
drying agent (Alves et al,. 2016). A combination of 10% maltodextrin and 5%
fructooligosaccharide has been considered to be the preferred carrier for drying fermented litchi
juice using L. plantarum (Kalita et al,. 2018). Colín-Cruz et al,. (2019) encapsulated by spray
drying, in temperature of 130oC, blackberry juice (BJ) and Lactobacillus acidophilus (LA) with
MD, GA, whey protein concentrate (WPC) and they mixtures in different proportions also
observed high survival of bacteria after spray drying. Also, they noticed that after 10 weeks of
storage the amount of bacteria decreased from 71-93 to 50-30%. Single probiotic cultivars was
encapsulated with MD or its mixture with GA at similar inlet air temperature (130-150oC) and a
high survival of bacteria was observed (Arepally and Goswami 2019, Arepally et al,. 2020).
However, authors used water medium not juice medium with lowered pH, like in our experiments.
4.2. General properties of powders

Characteristic of powders included dry matter, water activity and diameter of powders
which can tell us about future storage condition of powders and degradation by water adsorption.
What more colour of powder particles was tested to obtained correlation with pigment amount.
Colour perception can be also influenced by the morphology of the powder particles, which was
also tested.
Dry matter and water activity
It was observed that in all powders dry matter content were high (above 95%) and water
activity was below 0.25 (Table 1), independently from used bacteria type for fermentation process
and kind of carrier material. Similar values (for inlet air temperature 150oC) were observed by
Arepally and Goswami (2019) for spray dried probiotic culture Lactobacillus Acidophilus with
maltodextrin and gum Arabic as a carriers.

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Accepted Article It was not seen any correlation between tested carriers and bacteria type on those physical
parameters. However, it was observed correlation between dry matter content and water activity,
the highest the dry matter content the lowest water activity value (Table 1). As was stated high
moisture content (so low d.m.) can affect the microbial growth and physical properties of powders
(Guergoletto et al,. 2017). All powders during storage in typical conditions (20oC, humidity
bellow 40HR) would be stable and no microbial growth will be possible because of lower water
availability (Dantas et al,. 2018, da Silva Carvalho et al,. 2016).
After storage for 3 months decrease in dry matter and increase in water activity (Table 1)
were observed. Moreover, obtained values for powders are still considered as a safe.
Morphology and size of microparticles
All microcapsules of fermented beetroot powders had a typical for spray dried powders
morphology, regardless of the used bacteria and media type. It was observed spherical shape with
many wrinkles and holes, also damaged particles and high distribution of sizes was seen (Figure
S1 in supplementary file). No differences, except size, in morphology was observed. On the
particles outher morphology no bacteria strains were seen. However, after microbial analysis,
bacteria in amount from 2.31 ± 0.01 Log CFU/g to 3.90 ± 0.04 Log CFU/g was marked (Table 1)”,
so it was assumed that they survive inside the microcapsules. The same observation was made by
Rajabi et al,. (2015), Alves et al,. (2016) for probiotic orange juice, Guergoletto et al,. (2017) for
probiotic jucara pulp.
It was observed that amount of found colony of bacteria (Table 1) have influenced the size of the
particles, the lower the colony number (CFU) the higher the diameter d50 (Table 1). Moreover,
change of carrier material, for bacteria type L. brevis (LB) and L. plantarum (LP), caused
significant modification in diameter size. However, the experiments for bacteria types L.
fermentum (LF) and mixture of all bacteria types in proportion 1:1:1 showed that diameter after
addition of gum Arabic was lower in comparison to those with MD used as a carrier material. As
was stated by (Guergoletto et al,. 2012) that higher diameter size of particles can lead to an
increase in contact-time between the hot air and the material in processing, and consequently
lower bacteria amount, by its degradation during spray drying (Guergoletto et al,. 2017), what was
confirmed in presented research (Table 1).
Colour of fermented beetroot juice microparticles
Colour of food is important parameter, which is define as a main purpose for buying food.
It can be correlated with colourant content and by that prediction of antioxidant activity can be

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 done (Guergoletto et al,. 2017, Janiszewska 2014). Analysis of the powder colour coefficients
Accepted Article
showed that used for fermentation bacteria LAB and carrier type had significant impact on colour
parameters (Table 1).
It was seen reverse tendency for powder based on fermented beetroot juice by bacteria L.
fermentum (LF) and L MIX to those with L. brevis (LB) and L. plantarum (LP), independently
form carrier material. When MD as a carrier material was replaced in 1/3 by gum Arabic for
powders created with juice fermented by L. fermentum (LF) and L MIX increase for lightness and
decrease for redness and yellowness were observed. In contrast, in the case of powders based on
juice fermented by L. brevis (LB) and L. plantarum (LP), decrease for lightness, no changes for
redness and no clear trend for yellowness were observed (Table 1).
Change in bacteria type during fermentation process from L. brevis (LB) L. plantarum
(LP) L. fermentum (LF) caused statistically significant increase in lightness and yellowness, but
no clear trend for redness was seen (Table 1). Different observation was made for particles
contained mixtures of used bacteria, those particles was characterised by lover lightness and
yellowness but higher redness.
Those differences was linked to the size of particles and also number of bacteria present
in particles. For particle created with bacteria L. fermentum (LF) or L MIX bigger diameter was
observed what can during measurement create different light angle and different read of colure.
That can be also a reason of obtained higher and in the most cases positive (b* +) values for those
powders.
It was observed there was a correlation between lightness and d50 (L* = 29,6174 +
0,6509* d50; r = 0,9531; p = 0.0000) and for yellowness and d50 (b* = 1.12d50 – 40.15, R = 0.786)
and redness and diameter d50 (a* = 17,7528 + 0,6989* d50; r = 0,8538; p = 0,00000).
No correlation between the content of red betalain pigment and red coefficient a* was
observed (red colorant = 17,5845 + 0,2461*a*; r = 0,2792; p = 0,1766); but correlation for
coefficient b* and yellow colourant was stated; (yellow colourant = -52,1887 + 0,7704*b*; r =
0,6251; p = 0,0011). It is in contrast for research made for non-fermented chokeberry juice
(Bednarska and Janiszewska-Turak 2020), for non-fermented beetroot juice colourants
encapsulated by Azeredo et al,. (2007) and anthocyanin pigments obtained by Jiménez-Aguilar et
al,. (2011).

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 4.3. Quantification of betalain and polyphenols
Accepted Article The highest content of red pigments was observed for microcapsules obtained by bacteria
type L. brevis (LB), the lowest for particles with bacteria L. fermentum (LF) and MD as a carrier.
For yellow pigments the highest content was observed in fermented beetroot particles with L MIX
and L. fermentum (LF) with MD:AG=2:1 as a carriers, the lowest was seen for sample L. brevis
(LB) with MD (Figure 1).
Change in carrier material only for powders containing bacteria L. fermentum (LF) and L
MIX caused statistically significant increase in red colorant content (Figure 1). Those powders had
bigger diameter and lower bacteria amount, and despite the statistical increase, the content of red
pigments in these powders was at an average level.
Similar values of betanin and vulgaxhanthin-I were observed also for spray dried non-
fermented beetroot powders (Janiszewska 2014, Henriette M. C. 2009) and for betalain from
cactus pear (Saenz et al,. 2009). Unfortunately, it was not found results for antioxidant compounds
for fermented beetroot powders for fermented juices.
Change in bacteria type during fermentation process from L. brevis (LB) L. plantarum
(LP) L. fermentum (LF) caused decrease in red colorant content and no clear trend for yellow
pigment was observed (Figure 1). Moreover, for bacteria mixture (L MIX) the average red
pigment content was seen, yellow pigment was at the highest level from all tested samples.
Analysis of polyphenol content did not showed any differences with changing carrier
material type and used bacteria type. The highest values of polyphenol content was observed for
powder containing bacteria L. brevis (LB), independently from carrier material type.
Conclusion

Analysis of obtained result showed that physical parameters of obtained powders were
similar and all powders would be stable (low water activity bellow < 0.25, high dry matter content
95-98%). It was observed that bacteria type has the main influence on the chemical properties and
amount of bacteria. The highest bacteria, polyphenols and betalain content were observed in
powders with LB as a starter. Some correlations were observed: correlation between colour
coefficients (L*, a*, b*) and diameter d50, correlation for colour coefficient b* and yellow
colourant content. However, the method of drying fermented juices still needs to be refined due to
the reduction in the microorganisms number during the drying process. Good opportunity could be

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 decreasing the outlet temperature. It could be obtained also by changing the humidity of the drying
Accepted Article
air.

Conflicts of interest

The authors declare that they have no conflict of interest.

Data Availability Statement

Research data are not shared

Ethical approval

Ethics approval was not required for this research.

Acknowledgement
The work was co-financed by a statutory activity subsidy from the Polish Ministry of Science and
Higher Education for the Institute of Food Sciences of Warsaw University of Life Sciences and
grant from National Science Centre (NCN) number 2019/03/X/NZ9/00388.
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 Bazaria, B. & Kumar, P. (2016b). Effect of whey protein concentrate as drying aid and drying parameters
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survival of Lactobacillus reuteri LR92 in fermented juçara pulp after dehydration by spray-drying

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Accepted Article and the influence on the physical properties of the powders obtained therefrom. Spray-dried
powders were produced using 10% of carrier agents, and analyses of cell viability, moisture
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 Kazimierczak, R., Siłakiewicz, A., Hallmann, E., Srednicka-Tober, D. & Rembiałkowska, E. (2016). Chemical
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 Rajabi, H., Ghorbani, M., Jafari, S. M., Sadeghi Mahoonak, A. & Rajabzadeh, G. (2015). Retention of saffron
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 Zielinska, M. & Waszkiewicz-Robak, B. (2015). Tradycyjne roślinne napoje fermentowane o działaniu
Accepted Article prozdrowotnym. Przemysł Fermentacyjny i Owocowo-Warzywny, 5.

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 
Accepted Article Fermented beetroot powders have a high content of bioactive components

 Fermented beetroot powders have low bacteria amount

 Method of spray drying fermented juices needs to be refined

Figure 1. Betanin and polyphenol content in tested powders

a, b ... - different indexes for individual series mean statistically significant differences for given
values at the level of p < 0.05

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Accepted Article Table 1. Microbial and physical properties of obtained fermented beetroot juice particles

Water activity
Dry matter
Bacteria number aw
d.m. Colour parameters Diameter
[log CFU/g] (-)
(%) d50
Sample Name xav±SD
(m)
Beetroot Beetroot
after L* a* b* xav±SD
juice powder after powder after after drying after drying after storage
storage xav±SD xav±SD xav±SD
drying storage
LB MD 3.59 ± 0.08b 3.55 ± 0.04c 95.0±0.3a 94.5±0.5a 0.25±0.01c 0.27±0.01c 43.0±0.0a 39.1±0.0c -18.4±0.0b 20.75±0.40a
7.79 ±0.06b
LB MD:GA=2:1 3.90 ± 0.04c 3.80 ± 0.05d 96.1±0.2c 95.4±0.2c 0.19±0.01ab 0.21±0.01ab 45.4±0.5b 38.3±1.3c -19.2±0.3a 25.13±1.20b
LP MD c
3.09 ± 0.04a 3.12 ± 0.08b 96.5±0.2cd 95.8±0.2cd 0.17±0.00a 0.19±0.00a 49.0±0.0c 39.2±0.1c -18.5±0.1b 20.75±0.40a
7.92 ± 0.05
LP MD:GA=2:1 3.69 ± 0.06b 3.59 ± 0.05c 96.0±0.0bc 95.4±0.0bc 0.20±0.01ab 0.22±0.01ab 48.8±0.2c 38.3±0.3c -17.5±0.2c 24.45±1.74b
LF MD 2.48 ± 0.12a 2.33 ± 0.06ab 95.1±0.1ab 94.8±0.1ab 0.20±0.00ab 0.21±0.00ab 51.1±0.3e 34.9±0.4b 2.7±0.1g 40.46±1.67e
6.30 ± 0.05a
LF MD:GA=2:1 2.99 ± 0.01a 2.91 ± 0.03ab 97.2±0.5d 96.7±0.5d 0.17±0.01a 0.19±0.01a 48.7±0.0c 31.7±0.0a 0.1±0.0e 30.51±0.29c
L MIX MD 2.31 ± 0.01a 2.13 ± 0.04a 95.0±0.1a 94.4±0.1a 0.22±0.01bc 0.23±0.01bc 49.7±0.1d 37.9±0.1c 1.0±0.0f 35.53±1.74d
6.73 ± 0.05a
L MIX MD:GA=2:1 2.79 ± 0.05a 2.50 ± 0.10ab 96.4±0.2cd 95.7±0.2cd 0.18±0.01a 0.20±0.01a 45.1±0.0b 32.1±0.0a -1.9±0.0d 23.87±0.15ab
a, b ... - different indexes for individual series mean statistically significant differences for each column at the level of p <0.05

abbreviations: LB -L. brevis, LP- L. plantarum, LF - L. fermentum, L MIX - combination of tested strains in the ratio of 1: 1: 1, MD-
maltodextrin, MD:GA=2:1 – maltodextrin with gum Arabic in proportion 2:1

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Accepted Article

ijfs_14915_f1.tif

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