Synbiotic functional drink from Jerusalem

artichoke juice fermented by probiotic

Lactobacillus plantarum PCS26

Darko Dimitrovski, Elena Velickova,

Maja Dimitrovska, Tomaz Langerholc &
Eleonora Winkelhausen

Journal of Food Science and

Technology

ISSN 0022-1155
Volume 53
Number 1

J Food Sci Technol (2016) 53:766-774

DOI 10.1007/s13197-015-2064-0

1 23
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 Author's personal copy
J Food Sci Technol (January 2016) 53(1):766–774
DOI 10.1007/s13197-015-2064-0

ORIGINAL ARTICLE

Synbiotic functional drink from Jerusalem artichoke juice

fermented by probiotic Lactobacillus plantarum PCS26
Darko Dimitrovski 1 & Elena Velickova 1 & Maja Dimitrovska 2 & Tomaz Langerholc 3 &
Eleonora Winkelhausen 1

Revised: 17 August 2015 / Accepted: 7 October 2015 / Published online: 25 October 2015
# Association of Food Scientists & Technologists (India) 2015

Abstract A probiotic strain Lactobacillus plantarum PCS26 blueberry juice showed that the 50/50 % v/v mixture would be
was used to ferment Jerusalem artichoke juice. Growth kinetics very well accepted by the consumers. Above 80 % of the panel-
of the bacterial strain was followed during juice fermentation ists would buy this drink, and over 60 % were willing to pay
both in flask and in laboratory fermentor. Jerusalem artichoke more for it. Culture survivability in the fermented juices during
showed to be an excellent source of nutrients for L. plantarum storage at 4–7 °C was assayed by the Weibullian model. The
PCS26 growth. The culture grew very well reaching more than product shelf-life was extended from 19.70 ± 0.50 days of pure
1010 cfu/ml in just 12 h. The pH changed from the initial 6.5 to Jerusalem artichoke juice to 35.7 ± 6.4 days of the mixture con-
4.6 at the end of fermentation. The culture hydrolyzed fructool- taining 30 % blueberry juice.
igosaccharides present in the Jerusalem artichoke juice, yielding
fructose which was presumably consumed along with the malic
Keywords Lactobacillus plantarum . Jerusalem artichoke .
acid as energy and carbon source. Lactic acid was the main
synbiotic beverage . probiotic . Weibullian model
metabolite produced in concentration of 4.6 g/L. Acetic and
succinic acid were also identified. Sensory evaluation of the
fermented Jerusalem artichoke juice and its mixtures with
Introduction
Research highlights
• Probiotic L. plantarum PCS26 was used to ferment Jerusalem artichoke Probiotic foods and beverages are gaining in popularity as
juice more and more consumers become aware of their health ben-
• The culture reached 1010 cfu/ml in just 12 h lowering the pH from 6.5 to 4.6
efits. In addition to dairy products, in particular yogurt, fruit
• L. plantarum PCS26 hydrolyzed the fructooligosaccharides present in the J.
artichoke juice and vegetable juices were found to be good vehicles to deliver
• Mixtures of fermented J. artichoke juice and blueberry juice showed good probiotic microorganisms. Therefore, the development of
sensory properties such products is one of the research priorities for food indus-
• Culture survivability was assessed by Weibullian model and shelf life of
try, particularly in developed countries, where demand for
36 days is expected
vegetarian probiotic products is boosting (Betoret et al.
* Darko Dimitrovski
2012). However, the survival of the health-conferring micro-
[email protected] organisms in these media remains a main challenge for man-
ufacturers. As reported, prebiotics, non-digestible ingredients
1
in foods, can improve the survivability of probiotics, live mi-
Department of Food Technology and Biotechnology, Faculty of
Technology and Metallurgy, Ss. Cyril and Methodius University,
croorganisms which confer health benefits on the hosts when
Rudjer Boskovic 16, 1000 Skopje, Macedonia consumed in adequate amounts (Rezaei et al. 2014). Further-
2
Institute of Public Health of the Republic of Macedonia, 50 Divizija
more, when probiotics and prebiotics had been used together,
6, 1000 Skopje, Macedonia a higher advantage to the host was noticed due to their syner-
3
Department of Microbiology, Biochemistry, Molecular Biology and
gistic action (Vitali et al. 2010). Like probiotics, prebiotics are
Biotechnology, Faculty of Agriculture and Life Sciences, University also involved in treatment of irritable bowel syndrome and
of Maribor, Pivola 10, 2311 Hoce, Slovenia inflammatory bowel diseases (Ghouri et al. 2014).
 Author's personal copy
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Jerusalem artichoke (Helianthus tuberosus L.) is a perenni- indigenous presence of inulin in the Jerusalem artichoke juice
al plant from the sunflower family Asteraceae cultivated in and the probiotic organism would result in a synergistic action
North America, Europe, Asia and Australia (Paseephol et al. on human health. The second goal was to scale up the fermen-
2007). Although by name it is a type of artichoke, neither it tation process and to evaluate the survivability of the culture
originates from Jerusalem nor does it belong to the same ge- using the Weibullian model.
nus widely known as artichoke plant. It comprises a great
nutritive value which comes primarily from its storage carbo-
hydrates, among which inulin is present in the highest con- Materials and methods
centrations (Jovanovic-Malinovska et al. 2014). When con-
sumed, inulin is not digested in the upper digestive tract, but Preparation of Jerusalem artichoke juice
rather fermented by native microflora of the colon. Therefore
it is considered as prebiotic which can affect both the compo- Jerusalem artichoke juice was prepared from fresh tubers ob-
sition and/or activity of the gastrointestinal microflora that tained from local green market. After washing and roughly
confers health benefits to host (Slavin 2013). Jerusalem arti- pealing, the tubers were cut into smaller pieces (1–2 cm3)
choke has low caloric value since it does not contain fats and and fed to the kitchen juicer. The juice was filtered through
starch. It is a good source of B-group vitamins, trace elements, cheese cloth to remove the debris and blanched in boiling
in particular iron, and is well known for decreasing the levels water for 2 min. The soluble solids were measured by refrac-
of blood cholesterol and triglycerides (Grela et al. 2014). tometer (around 20 % w/v) and adjusted to 10 % (w/v) by
There is a large number of publications dealing with the fer- dilution (approximately 1:2) with sterile distilled water.
mentation of Jerusalem artichoke and generating different
products, starting from organic compounds for analytical pur- Strain and cultivation
poses to single-cell proteins (Gao et al. 2007). Jerusalem arti-
choke has been often used as substrate for bio-fuel production Lactobacillus plantarum PCS26 (Deposited at Microbial
like ethanol and butanol by yeast fermentation (Sarchami and Strain Collection of Latvia, accession number: PCS 26 (P
Rehmann 2014). It is also known as a raw material for inulin, 975)) (PathogenCombat 2011) was kept at −20 °C and revi-
fructose and oligosaccharides production in the medical and talized by overnight growth in de Man, Rogosa and Sharpe
pharmaceutical industry (Jovanovic-Malinovska et al. 2015). (MRS) broth (Merck, Whitehouse station, New Jersey, USA)
Concerning its raw consumption, it is still mostly used as at 37 °C using semi-anaerobic conditions. Overnight
animal feed. However, due to its dietary properties, the pres- preculture of L. plantarum PCS26 incubated in MRS broth
ence of Jerusalem artichoke in human diets gradually evolves. at 37 °C was used as inoculum. Flask fermentations were
Although lactic acid fermentation of Jerusalem artichoke carried out with 300 mL Jerusalem artichoke juice in a
tubers dates back to 1942 (Andersen and Greaves 1942), it 500-mL Erlenmeyer flask placed on a rotary shaker
still remains an interesting topic. Dao et al. (2013) evaluated (120 rpm) at 37 °C for about 35 h. Inoculation with 30 μL
the application of enzymes for inulin hydrolysis and of the probiotic preculture was sufficient to result into initial
fermented the hydrolyzed Jerusalem artichoke tubers by viable count of about 105 cfu/mL in all fermentations.
Pediococcus acidilactici DQ2 to generate lactic acid with high Scaling up of the fermentation of the Jerusalem artichoke
productivity. Most of the lactic acid bacterial fermentations of juice was carried out in 1-L laboratory fermentor (B. Braun
Jerusalem artichoke tubers were performed after inulin hydro- Biotech International, Melsungen, Germany). The sterilized
lysis to reducing sugars and by supplementing the media with fermentor was filled with 800 mL Jerusalem artichoke juice
growth promoting components (Cheng et al. 2014). In con- and inoculated with 150 μL of overnight MRS preculture to
trast, Choi et al. (2012) demonstrated that certain reach initial viable count of 105 cfu/mL.
Lactobacillus species can efficiently ferment fructooligosac- All fermentations were performed in triplicates. Samples
charides with degree of polymerization up to 13 without any were taken aseptically at appropriate time intervals to evaluate
pretreatment. the viable count and pH value.
Lactobacillus plantarum PCS26 was isolated from tradi-
tional Slovenian cheese and studied for its probiotic proper- Sample analysis
ties. Its extracellular metabolites showed several positive ef-
fects on the gut epithelial cells during in vitro tests The number of viable cells was measured by standard
(Dimitrovski et al. 2014). Therefore, the primary aim of this colony counting method. After a series of appropriate
study was to ferment Jerusalem artichoke juice, without any dilutions, samples were plated on MRS agar and incubated
pretreatments or supplementations, with the probiotic bacteri- at 37 °C for 48 h before colony counting. pH of the fermen-
um, Lactobacillus plantarum PCS26 and to create a functional tation broth was measured with a pH-meter (Sartorius PB-11,
synbiotic drink with acceptable sensory properties. The Göttingen, Germany).
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768 J Food Sci Technol (January 2016) 53(1):766–774

Agilent 1200 High Performance Liquid Chromatography first set of test consisted of evaluating the taste, color and
(HPLC-Agilent Technologies, Inc., Santa Clara, USA) was smell of the pure JA juice and its mixtures with a wide variety
used for quantification of reducing sugars in the samples. Sep- of commercial 100 % vegetable and fruit juices, such as: beet
aration of the sugars was done on Supelcosil LC-NH2 column, root, carrot, apple, orange, lemon, blueberry, pineapple, cher-
250 × 4.6 mm, 5 μm particle size, (Supelco analytical, Sigma ry, raspberry and pear, all purchased from the local grocery
Aldrich Group, Taufkirchen, Germany) using isocratic mobile store. The tasting and the evaluation was done by 3 trained
phase acetonitrile/water =75/25 (v/v) at 40 °C (Muntean and judges, food engineers, working on development of new com-
Muntean 2010). Refractive index detector, thermostated at mercial drinks. After the first set of tests, the number of pos-
40 °C, fed the software (Agilent ChemStation) with data for sible commercial juices for mixing was reduced to 5. After
analysis. The run time was 15 min at 1.0 mL/min flow of the tasting different ratios of blueberry, pineapple, cherry, raspber-
mobile phase. ry and beet root juice with the JA juice, the blueberry (100 %
Identification of inulin (degree of polymerization >10) and fruit) was chosen as a best option for the mixtures.
other fructooligosaccharides with lower degree of polymeri- In the second part, mixtures of fermented Jerusalem arti-
zation was carried out using thin layer chromatography ac- choke juice with blueberry juice (100 % fruit) were evaluated
cording to Jovanovic-Malinovska et al. (2014) with minor for their sensory quality in a standardized test room. Three
adjustments. The samples were diluted (1:5) and applied in mixtures were prepared containing different percentage of Je-
quantity of 2 μL on a silica gel 60 F254 plates (aluminium rusalem artichoke juice: 50 %, 60 % and 70 %. Each of them
sheets 20 × 20 cm, Merck, Darmstadt, Germany). Aqueous was served in a randomized order in a white plastic cup la-
solutions of 1 % w/v inulin, fructose, glucose and sucrose were beled with three digit random number. The panel consisted of
used as standards. Solvent system acetonitrile/water (75:25 v/ 19 random panelists selected from a larger group of possible
v) was used as mobile phase to separate the carbohydrates at testers due to their declaration as frequent juice drinkers will-
room temperature. Since this mobile phase did not result into ing to pay more than the average cost for juice, 1.5 ± 0.5 euros/
satisfactory separation of fructose and glucose, another mobile L, for products that satisfy their needs and taste. Among the 19
phase with higher concentration of acetonitrile (85:15 v/v) was panelists, 10 were women and 9 were men in the age range
also tested, and it successfully separated the monosaccharides. from 30 to 60 years. Before the tasting, the juices were intro-
Unfortunately, with this mobile phase the separation of fruc- duced to them and they were briefed on the attributes that
tooligosaccharides deteriorated. Therefore, these two mobile should be evaluated. The acceptance of color, smell and taste
phases of acetonitrile/water, 75:25 v/v and 85:15 v/v, were as well as overall acceptance were graded from 1 (not accept-
used on two separate plates to analyze the components. Spots able) to 5 (very good). Afterwards, two more questions with
were visualized by spraying the plates with 5 % w/v phenol in offered answers followed: 1. would you buy this product?
10 % v/v sulphuric acid followed by heating at 120 °C for (yes/no) and 2. are you ready to pay more for it than the
5 min. The analysis was done in triplicate. regular price for juice? (yes/no).
The concentration of the organic acids was determined
using a Shimadzu Prominence Liquid Chromatography
(Shimadzu corp., Kyoto, Japan). The separation of the com- L. plantarum PCS26 viability during storage of fermented
pounds was achieved at 55 °C using Aminex HPX-87 H col- Jerusalem artichoke juice
umn, 300 mm × 7.8 mm ID, 5 μm particle size (Bio-Rad
Laboratories, California, USA) according to the manufacturer The pure fermented Jerusalem artichoke juice and its mixtures
instructions. Isocratic elution was applied with a mobile phase with blueberry juice were refrigerated (4-7 °C) at the moment
consisting of 2.5 mM H2SO4 at a flow rate of 0.6 mL/min. The when L. plantarum PCS26 reached the maximal cell density.
wavelength selected for detection purposes was 214 nm and Samples were collected at regular time intervals during the
the measured data were processed by Class VP 7.3 software. following 16 days and viable count was determined. Weibull
The concentrations of the measured components were cal- distribution, nonlinear survival model (Equation 1) (van
culated from the peak areas using appropriate standard curves Boekel 2009), was fitted to the data and used to estimate the
generated with external standards. The presented data are av- storage time by which the probiotic juice would retain viable
erage values of three independent measurements and the stan- count above 106 cfu/mL.
dard deviations are shown as error bars.
N
log ¼ −b ⋅ t n ð1Þ
Sensory evaluation No
N and No represent the viable count at time t and at t = 0,
Sensory evaluation of the fermented Jerusalem artichoke  β w
juices was performed in two parts. In the first part the prelim- respectively; b ¼ 2:303
1
⋅ α1w ; and n = βw where αw and βw
inary sensory analysis was carried out in two sets of tests. The are the two parameters of the distributions: αw is a scale
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parameter (a characteristic time) and βw is the so-called shape start of the fermentation. The culture grew 2 log cycles in 7 h
parameter. reaching 3.1 × 1011 cfu/mL with μ of 0.24 h−1. After relatively
short stationary phase of 4 h, ending in total fermentation time
Statistical analysis of 23.5 h, the culture started decreasing its viable count. Dur-
ing the second growth phase a rapid and pronounced decrease
The growth curves were plotted using representative results of pH was noticed reaching pH 5.3. The pH decreasing trend
from several fermentations where data were average values of continued all through the stationary phase at a slower rate,
triplicate measurement of the samples. The presented data of attaining final value of pH 4.7 at the end of the fermentation.
the HPLC analysis are average values of three independent The diauxic growth of L. plantarum when cultivated on
fermentations. The standard deviations are shown as error complex carbohydrates was also noticed by Hasan and Durr
bars. Sensory evaluation data were statistically analyzed using (1974). Goh et al. (2007) observed the same growth pattern for
Anova with Tukey post-hoc test (SPSS software, IBM Corpo- L. paracasei in the presence of fructooligosaccharides and
ration, Armonk, New York, USA) for determination of statis- limited amount of glucose in the medium (0.1 %). They con-
tically significant difference between the two populations of cluded that oligosaccharides are hydrolyzed extracellularly
values at 95 % confidence interval. The equality of variances with the cell wall associated enzyme β-fructosidase, induced
was checked by the Levine’s test. Binary logistic regression by the presence of inulin, sucrose and fructose in the medium.
was used to test which sensory attribute influenced the panel- Recently, several studies were published about different
ists' decision the most, in terms of buying and paying more for Lactobacillus species that utilize fructooligosaccharides as en-
the synbiotic drink compared to regular juices. ergy source (Endo et al. 2012; Zubaidah 2013). The fructool-
igosaccharides consumption by L. plantarum WCFS1 was
studied in detail by Saulnier et al. (2007). They found that
the main mechanism includes phosphoenolpyruvate transport
Results and discussion
system, β-fructofuranosidase, fructokinase and α-glucosi-
dase. Trisaccharide 1-kestose was preferentially utilized in
Flask fermentation
comparison to the tetrasaccharide nystose and the pentasac-
charide fructofuranosylnystose.
Cell concentration and pH
Inulin and other fructooligosaccharides
The viable count and pH development during the L. plantarum
PCS26 growth in Jerusalem artichoke juice is presented in
The analysis of the Jerusalem artichoke juice by thin layer
Fig. 1. Two growth phases can be noticed separated by a very
chromatography indicated a complex composition of the car-
short lag phase between 11.5 and 12.5 h from the start of the
bohydrates (Fig. 2). As can be noticed from Fig. 2a (lines 1
fermentation. This, what seems to be a diauxic growth, started
and 2), Jerusalem artichoke juice consists of sucrose, fructo-
after 7 h lag phase during which the culture adapted to the
oligosaccharides of different degree of polymerization and
environment. Throughout the first growth phase the culture
inulin. Minor amounts of fructose and glucose, not detectable
grew 4 log cycles in 4.5 h, reaching 6.3 × 109 cfu/mL with
by thin layer chromatography, might be present as well. There
specific growth rate (μ) of 0.85 h−1. Minor change of the pH
was no major change in the carbohydrate composition as a
value, from 6.7 to 6.2, was noticed at this phase. The second
result of the blanching process. Similar results for the carbo-
growth phase was characterized by significantly slower
hydrate composition of the Jerusalem artichoke were obtained
change of the viable count and ended after 19.5 h from the
by Jovanovic-Malinovska et al. (2014). The samples of dilut-
ed Jerusalem artichoke juice (10 % w/v), fermenting Jerusalem
viable count pH
14.0 7.5
artichoke juice during the exponential phase and fermenting
7.0
Jerusalem artichoke juice during stationary phase were ana-
12.0 lyzed in lines 3, 4 and 5, respectively. It can be observed that
logN (cfu/mL)

6.5
10.0 6.0
during the fermentation certain amounts of monosaccharides
pH

were generated. To separate fructose and glucose, the latter

8.0 5.5
three samples (lines 3, 4 and 5) were additionally analyzed
5.0
6.0 using solvent system acetonitrile/water in ratio 85/15 v/v. The
4.5
resulting thin layer chromatogram revealed that mostly fruc-
4.0 4.0
0.0 10.0 20.0 30.0 tose was generated (Fig. 2b).
Time (h) These results suggested that L. plantarum PCS26 possesses
Fig. 1 Bacterial viable count and pH during flask fermentation of an extracellular enzymatic system able to hydrolyze the fruc-
Jerusalem artichoke juice by L. plantarum PCS26 tooligosaccharides into their building monomers. This finding
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770 J Food Sci Technol (January 2016) 53(1):766–774

Malic acid Lactic acid

6.0

5.0

Concentration (g/L)
4.0

3.0

2.0

1.0

0.0
0.0 10.0 20.0 30.0 40.0
Time (h)
Fig. 3 Malic and lactic acid concentration during the fermentation of
Jerusalem artichoke juice by L. plantarum PCS26

present in the Jerusalem artichoke juice in total of 2.1 ± 0.2 g/

L, started decreasing after 11.5 h, while the concentration of
lactic acid started increasing. The onset of these changes was
concomitant with the initiation of the second growth phase of
L. plantarum PCS26. After 21 h, the concentration of malic
acid was below the detection limit of the analytical technique
whereas lactic acid sustained its increase until the 24th h
reaching concentration of 4.6 ± 0.3 g/L. Theoretically, taking
into account that during malolactic conversion 1 mol of malic
acid converts into 1 mol of lactic acid, approximately 1.6 g/L
of lactic acid could be generated by malolactic conversion
alone. The remaining 3.0 g/L of lactic acid were presumably
produced by consumption of the hydrolyzed fructooligosac-
charides. The metabolism of fructooligosaccharides as energy
and carbon sources can be undertaken by two major pathways:
glycolysis (Embden-Meyerhof pathway), which is mostly
used by the homofermentative lactic acid bacteria, and 6-
phosphogluconate/phosphoketolase pathway mostly used by
the heterofermentative bacteria (Axelsson 1998).
Other components were also detected during the fermenta-
Fig. 2 Thin layer chromatography of Jerusalem artichoke juice carried tion, of which acetic and succinic acid were identified. Their
out with two acetonitrile/water solvent systems: a 75/25 v/v b 85/15 v/v. 1 concentrations, however, were below the quantification limits.
– before blanching, 2 – after blanching, 3 – after dilution, 4 – during
The production of succinic acid is often found in
fermentation (exponential phase), 5- during fermentation (stationary
phase). F- fructose, G – glucose, S – sucrose, DP – degree of heterofermentative lactobacilli due to the metabolism of the
polymerization terminal glucose in inulin and the citrate (Axelsson 1998;
Kaneuchi et al. 1988). These acids could be responsible for
is in agreement with a behavior of other Lactobacillus species the small pH drop from 6.7 to 6.2 before the 11.5th h, when no
which could ferment Jerusalem artichoke tubers without acid- lactic acid production was observed.
ic or enzymatic inulin hydrolysis prior to fermentation (Choi Cheng et al. (2014) tested the fermentative activity of
et al. 2012). The preference between different fructooligosac- L. plantarum CX-15 on Jerusalem artichoke extracts. They
charides such as 1-kestose (GF2) and nystose (GF3) was stud- succeeded in increasing the cell growth and producing lactic
ied for different Lactobacillus species by Endo et al. (2012). acid in concentration of approximately 12 g/L at the end of the
fermentation. This was achieved by medium supplementation
with Mn+2, which is an essential growth factor for lactic acid
Organic acids production during fermentations bacteria. Metabolites developed during fermentation of Jeru-
salem artichoke juice with different Lactobacillus strains
The concentrations of malic and lactic acid during the fermen- (L. plantarum, L. paracasei, L. casei, L. rhamnosus and
tation of Jerusalem artichoke juice by L. plantarum PCS26 are L. curvatus) were analyzed by Zalán et al. (2011). They found
depicted in Fig. 3. The concentration of malic acid, initially only three organic acids produced by the tested strains: lactic,
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viable count pH JA juice/BJ - 50/50 JA juice/BJ - 60/40 JA juice/BJ - 70/30

10.0 7.0 100.0

Panelists that answered YES (%)

9.0 6.5
80.0
8.0 6.0
logN (cfu/mL)

60.0

pH
7.0 5.5

6.0 5.0 40.0

5.0 4.5
20.0
4.0 4.0
0.0 5.0 10.0 15.0 20.0 0.0
Time (h) I would buy thisproduct I would pay more for it
Fig. 4 Bacterial viable count and pH during the fermentation of Fig. 5 Panelists that would buy and pay more for the drink prepared by
Jerusalem artichoke juice by L. plantarum PCS26 in a laboratory mixing fermented Jerusalem artichoke juice with blueberry juice
fermentor
sharply from 6.4 to 5.4 within 3 h of the first growth phase
acetic and succinic acid. All strains produced lactic acid, one (5th to 8th h).
strain did not produce acetic acid (L. paracasei subsp. casei It can be concluded that the culture in the fermentor had
SF1), and three strains produced succinic acid including decreased growth when compared to the flask fermentation
L. plantarum 2142. Acetoin and diacetyl were also found to resulting into lower growth rates and lower maximal viable
be produced in different concentrations. These two metabo- count. The results suggested that the production of lactic acid,
lites are produced in small concentrations during the citrate as the major metabolite influencing the pH change, was also
metabolism and they are very important flavoring components affected by the scaled up conditions in the reactor. It has been
in the fermented products (Bassit et al. 1993). frequently reported that scaling up fails to achieve the same
results concerning the biomass yield and other metabolites
Fermentation in a laboratory bioreactor associated with its growth (Enfors et al. 2001). Part of the
mechanisms responsible for these changes are the different
Scaling up of the Jerusalem artichoke juice fermentation for systems of agitation and heating responsible for the homoge-
producing a synbiotic drink was conducted as a batch process neity and the heat distribution in the fermentor (Hewitt and
in 1-L laboratory bioreactor. The viable count and pH changes Nienow 2007).
during the fermentation of the juice are given in Fig. 4. Al-
though the overall shape of the growth curve was similar, the Sensory evaluation of fermented Jerusalem artichoke
growth kinetics was different from flask fermentation. After juices
5 h of lag phase the culture started increasing the viable count
reaching 5.0 × 108 cfu/mL at the 12th h from the beginning of After preliminary sensory analysis (data not shown) it was
the fermentation. This growth of three log cycles was also inferred that product properties such as color, smell and taste
divided into two growth phases observed by the change in of the Jerusalem artichoke juice need improvement. There-
the slope (specific growth rate) at the 8th h. Specific growth fore, it was decided to mix the fermented Jerusalem artichoke
rates of 0.65 h−1 and 0.30 h−1 were calculated for the first and juice with a fruit or vegetable juice that will cover up some of
the second growth phase. The stationary phase lasted 2 h (12th its earth-like flavors and change the indigenous brown color
- 14th h), and afterwards the cell concentration declined. into a more attractive one. The presence of lactic acid in the
Unlike flask fermentation where pH had a decreasing trend fermented juice was not mentioned as off-taste by any of the
throughоut the diauxic growth and the stationary phase of the panelists. After several different juices (blueberry, pineapple,
bacterium, the pH value in the fermentor broth dropped cherry, raspberry, beet root) had been tested, blueberry juice

Table 1 Intensity of sensory

attributes of fermented Jerusalem Fermented Jerusalem artichoke Color Smell Taste Overall acceptance
artichoke juice mixed with juice/blueberry juice (%, v/v)
different concentrations of
blueberry juice* 0/100 4.98 ± 0.71a 4.80 ± 0.95a 4.01 ± 0.79a 4.22 ± 0.91a
a
50/50 4.68 ± 0.67 4.11 ± 1.02b 4.17 ± 0.86a 3.97 ± 0.86a
60/40 4.16 ± 0.90ab 3.72 ± 0.96b 3.78 ± 1.06ab 3.42 ± 0.93ab
70/30 3.74 ± 0.99b 2.78 ± 1.06c 3.17 ± 1.25b 2.75 ± 1.21b

*the reported data are mean values ± standard deviation of 19 panelists. Different letters in a column designate
significantly different means by Tukey test (p < 0.05)
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772 J Food Sci Technol (January 2016) 53(1):766–774

a experimental data Weibullian model

60 % were ready to pay more compared to regular fruit juice.
1.0
It was interesting to notice smaller differences between the
0.0 panelists who would buy the other two mixtures and those
0.0 5.0 10.0 15.0 20.0 who would be willing to pay more for them. Around 63 %
-1.0 of the panelists stated that they would buy the drink with 40 %
log N/No

blueberry juice and 58 % would be prepared to pay more for it.

-2.0
For the mixture with 30 % blueberry juice, only 21 % of the
-3.0 panelists would buy and pay more for it. It can be assumed
that for this group the health benefits of the drink were more
-4.0
Time (d) important than its sensory attributes and price.
From the statistical results of the binary logistic regression
it was evident that only taste had statistically significant influ-
b JA juice/BJ - 50/50 JA juice/BJ - 60/40 JA juice/BJ - 70/30
ence on the willingness of the panelists to buy and pay more
0.5 for the mixed juices. Color and smell were statistically not
0.0
important in terms of influencing panelists' choice. It can be
0.0 2.0 4.0 6.0 8.0 10.0 concluded that the added blueberry juice could cover earth-
-0.5 like flavor and brownish color of Jerusalem artichoke
log N/No

fermented juice, providing a synbiotic drink that could be well

-1.0
accepted by consumers.
-1.5
L. plantarum PCS26 viability during cold storage
-2.0
Time (d)
The survivability of the probiotic bacterium L. plantarum
Fig. 6 Weibullian model fit to the viable count data of L. plantarum
PCS26 cells in a Jerusalem artichoke juice and b mixed drinks prepared PCS26 during cold storage of fermented Jerusalem artichoke
from Jerusalem artichoke (JA) juice and blueberry juice (BJ), during juice and its mixtures with blueberry juice is displayed in
refrigerated storage at 4–7 °C Fig. 6. The Weibullian model for the survival of the culture
was fitted to the experimental data, and the parameters n and b
(100 % fruit) was selected as the best option. The results of the were determined by least-squares method. The kinetic param-
sensory evaluation of the blueberry juice and the three mix- eters as well as the storage time (tst) estimates during which
tures containing 50 %, 60 % and 70 % fermented Jerusalem the probiotic concentration would stay above 106 cfu/mL, are
artichoke juice are presented in Table 1. The addition of blue- presented in Table 2. As can be noticed, the drink prepared by
berry juice enhanced the overall acceptance of the juices. The mixing the fermented Jerusalem artichoke juice with blueber-
mixture with 50 % blueberry juice had significantly higher ry juice in ratio 70/30 % v/v, would have the longest storage
grades for all attributes compared with the mixture containing time of 35.7 ± 6.4 days. Blueberry juice, with pH 3.0, contrib-
30 % blueberry juice (p < 0.05). This is a good indication that uted to decreasing the initial pH value of the fermented Jeru-
the mixture with 50 % blueberry juice, having grades of 4 and salem artichoke juice (pH 5) into 3.8, 4.1 and 4.3 in the mix-
above, could be well accepted by consumers. tures with 50 %, 40 %, and 30 % blueberry juice, respectively.
The readiness of the panelists to buy the synbiotic mixed Some components in the juices, like proteins, dietary fibers
juices is presented in Fig. 5. Bearing in mind the fermentation and metabolizable sugars can enhance the culture survival
process, it is believed that the price for these drinks would be while others, like phenolic compounds, can have a negative
higher than the price of any regular commercial juice. Most of influence on survival during cold storage (Corcoran et al.
the panelists chose the 50/50 mixture as the one they would 2005; Nualkaekul and Charalampopoulos 2011). The pres-
preferably buy over the other mixtures. Over 80 % of the ence of acids and the pH drop have also been found to signif-
panelists answered that they would buy this drink and above icantly increase the sensitivity of probiotic cultures in fruit and

Table 2 Parameters of the Weibullian models for different Jerusalem artichoke juices

Juice/parameters b (day) n tst (day)

Fermented Jerusalem artichoke juice 0.05 ± 0.01 1.55 ± 0.09 19.70 ± 0.50
Fermented Jerusalem artichoke juice/blueberry juice, (%v/v) 70/30 0.04 ± 0.01 1.15 ± 0.13 35.70 ± 6.40
60/40 0.03 ± 0.01 1.30 ± 0.14 22.50 ± 2.70
50/50 0.04 ± 0.01 1.64 ± 0.10 10.50 ± 0.24
 Author's personal copy
J Food Sci Technol (January 2016) 53(1):766–774 773

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