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Production of probiotic cabbage juice by lactic acid bacteria

Article  in  Bioresource Technology · September 2006

DOI: 10.1016/j.biortech.2005.06.018 · Source: PubMed

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 Bioresource Technology 97 (2006) 1427–1430

Production of probiotic cabbage juice by lactic acid bacteria

Kyung Young Yoon, Edward E. Woodams, Yong D. Hang *

Department of Food Science and Technology, Cornell University, Geneva, NY 14456, United States

Received 29 March 2004; received in revised form 14 June 2005; accepted 30 June 2005
Available online 24 August 2005

Abstract

Research was undertaken to determine the suitability of cabbage as a raw material for production of probiotic cabbage juice by
lactic acid bacteria (Lactobacillus plantarum C3, Lactobacillus casei A4, and Lactobacillus delbrueckii D7). Cabbage juice was inoc-
ulated with a 24-h-old lactic culture and incubated at 30 C. Changes in pH, acidity, sugar content, and viable cell counts during
fermentation under controlled conditions were monitored. L. casei, L. delbrueckii, and L. plantarum grew well on cabbage juice
and reached nearly 10 · 108 CFU/mL after 48 h of fermentation at 30 C. L. casei, however, produced a smaller amount of titratable
acidity expressed as lactic acid than L. delbrueckii or L. plantarum. After 4 weeks of cold storage at 4 C, the viable cell counts of L.
plantarum and L. delbrueckii were still 4.1 · 107 and 4.5 · 105 mL 1, respectively. L. casei did not survive the low pH and high acid-
ity conditions in fermented cabbage juice and lost cell viability completely after 2 weeks of cold storage at 4 C. Fermented cabbage
juice could serve as a healthy beverage for vegetarians and lactose-allergic consumers.
 2005 Elsevier Ltd. All rights reserved.

Keywords: Cabbage juice; Probiotic; Lactic acid bacteria; Lactobacillus casei; Lactobacillus delbrueckii; Lactobacillus plantarum

1. Introduction (Heenan et al., 2002). Traditionally, probiotics have been

Probiotics are defined as live microbial feed supple-
ment that beneficially affects the host by improving its
intestinal balance (Fuller, 1989). Most probiotic microor-
ganisms are lactic acid bacteria such as Lactobacillus
plantarum, Lactobacillus casei, Lactobacillus acidophilus,
and Streptococcus lactis (Sindhu and Khetarpaul, 2001).
Research has shown that addition of probiotics to food
provides several health benefits including reduction in
the level of serum cholesterol, improved gastrointestinal
function, enhanced immune system, and lower risk of co-
lon cancer (Berner and OÕDonnell, 1998; Rafter, 2003;
Saarela et al., 2002; McNaught and MacFie, 2001). Lac-
tic acid bacteria are commercially used as starter cultures
for the manufacture of dairy-based probiotic foods
*
Corresponding author. Tel.: +1 315 787 2265; fax: +1 325 787
2284.
E-mail address: ydh1@cornell.edu (Y.D. Hang).
added to yogurt and other fermented dairy products, but
lactose intolerance and the cholesterol content are two
drawbacks related to their consumption. In recent years,
consumer demand for non-dairy-based probiotic prod-
ucts has increased, and probiotics have been incorpo-
rated into drinks as well as marketed as supplements in
the form of tablets, capsules, and freeze–dried prepara-
tions (Shah, 2001). Fruits and vegetables are rich in func-
tional food components such as minerals, vitamins,
dietary fibers, and antioxidants (phytochemicals). Fur-
thermore, fruits and vegetables do not contain any dairy
allergens that might prevent usage by certain segments of
the population (Luckow and Delahunty, 2004).
Cabbage is a cruciferous vegetable, which is rich in
minerals, vitamin C, dietary fibers, and especially phyto-
chemicals (Chu et al., 2002). The objective of this study
was to determine the suitability of cabbage as a raw
material for production of probiotic cabbage juice by
probiotic lactic acid bacteria.

0960-8524/$ - see front matter  2005 Elsevier Ltd. All rights reserved.
doi:10.1016/j.biortech.2005.06.018
1428 K.Y. Yoon et al. / Bioresource Technology 97 (2006) 1427–1430

2. Methods USA). The values within rows that have no common

superscript are significantly different (p < 0.05) accord-
Cabbage (Brassica oleracea L. var. capitata L.) was ing to DuncanÕs multiple range test (SAS Institute, Cary,
purchased from a local store and kept at 4 C prior to NC, USA). Any two means not marked by the same
use. Cabbage juice was obtained with a Loomis press superscript (for example, a and b or b and c within rows)
operated at 2000 psi and sterilized for 15 min at 121 C. are significantly different (p < 0.05). Any two means
marked by the same superscript (for example, a and a
2.1. Probiotic lactic acid cultures or b and b within rows) are not significantly different
(p < 0.05).
Lactobacillus casei A4, Lactobacillus debrueckii D7,
and Lactobacillus plantarum C3 were obtained from
the New York State Agricultural Experiment Station 3. Results and discussion
Culture Collection, Geneva, New York. The cultures
were grown at 30 C for 24 h in MRS broth (dextrose All the three species of lactic acid bacteria, L. casei,
20.0 g/L; meat peptone 10.0 g/L; beef extract 10.0 g/L; L. delbrueckii, and L. plantarum, were found capable
yeast extract 5.0 g/L; sodium acetate 5.0 g/L; disodium of growing well on sterilized cabbage juice without
phosphate 2.0 g/L; ammonium citrate 2.0 g/L; tween nutrient supplementation. The time courses of lactic
80 1.0 g/L; magnesium sulfate 0.1 g/L, manganese sul- acid fermentation of cabbage juice by L. casei, L. plan-
fate 0.05 g/L). tarum, and L. delbrueckii are presented in Tables 1–3,
respectively. L. casei, L. plantarum and L. delbrueckii
2.2. Fermentation of probiotic cabbage juice grew rapidly on cabbage juice and reached nearly
10 · 108 CFU/mL after 48 h of fermentation at 30 C.
Fermentation experiments were conducted in test Extending the fermentation beyond 48 h did not result
tubes (25 · 200 mm), each containing 40 mL of sterile in a significant increase in the viable cell counts of lactic
cabbage juice. All samples were inoculated with a 24-h acid bacteria. Both L. plantarum and L. delbrueckii pro-
culture (>105 CFU/mL) and incubated at 30 C for duced significantly more titratable acidity expressed as
72 h. Samples were taken at 0, 24, 48, and 72 h for chem- lactic acid than L. casei. For example, L. plantarum
ical and microbiological analyses. and L. delbrueckii produced nearly 1% titratable acidity
expressed as lactic acid after 72 h of fermentation
2.3. Effect of cold storage on cell viability in probiotic at 30 C. Under similar growth conditions, L. casei
cabbage juice
Table 1
After 72 h of fermentation at 30 C, the fermented Time course of lactic fermentation of cabbage juice by Lactobacillus
samples were stored at 4 C for 4 weeks. Samples were casei
taken at weekly intervals, and the viability of probiotic Time pH Acidity Sugar CFU/mL
cultures in probiotic cabbage juice was determined and (h) (% lactic acid) (mg/mL)
expressed as colony forming units (CFU/mL). 0 5.0 ± 0.1a 0.11 ± 0.01d 45.6 ± 2.5a 3.0 ± 0.2 · 106a
24 3.7 ± 0.0b 0.38 ± 0.01c 41.7 ± 1.4b 6.3 ± 0.0 · 108b
2.4. Chemical and microbiological analyses 48 3.4 ± 0.0c 0.6 ± 0.03b 39.5 ± 1.8bc 12 ± 0.0 · 108c
72 3.4 ± 0.1c 0.74 ± 0.02a 36.5 ± 1.9c 11 ± 0.1 · 108c
The pH of probiotic cabbage juice was measured Means and standard deviations for n = 3. The experimental values
with a pH meter. Total acidity, expressed as percent lac- within rows that have no common superscript are significantly different
tic acid, was determined by titrating with 0.02 N NaOH (p < 0.05) according to DuncanÕs multiple test range.
to pH 8.2. Sugar content was analyzed as glucose by the
phenol sulfuric acid method of Dubios et al. (1956). Via-
ble cell counts (CFU/mL) were determined by the stan- Table 2
dard plate method with Lactobacilli MRS medium after Time course of lactic fermentation of cabbage juice by Lactobacillus
plantarum
48 h of incubation at 30 C.
Time pH Acidity Sugar CFU/mL
(h) (% lactic acid) (mg/mL)
2.5. Statistical analysis
0 5.8 ± 0.0a 0.12 ± 0.0a 35.08 ± 0.09a 8.0 ± 6.26 · 105a
24 4.8 ± 0.2b 0.23 ± 0.06b 37.10 ± 0.39a 7.7 ± 3.41 · 108b
All experiments were carried out in triplicate, and 48 3.6 ± 0.0c 0.76 ± 0.03c 36.44 ± 3.77a 15.3 ± 0.92 · 108c
each sample was analyzed in duplicate. The results are 72 3.6 ± 0.0c 0.97 ± 0.03d 19.33 ± 1.04b 17.5 ± 7.05 · 108c
expressed as mean ± S.D. (standard deviation). The Means and standard deviations for n = 3. The experimental values
SAS statistical computer package was used to analyze within rows that have no common superscript are significantly different
the experimental data (SAS Institute, Cary, NC, (p < 0.05) according to DuncanÕs multiple test range.
 K.Y. Yoon et al. / Bioresource Technology 97 (2006) 1427–1430 1429

Table 3 (Shah, 2001). Therefore, the viability of the lactic cul-

Time course of lactic fermentation of cabbage juice by Lactobacillus tures is the most important factor during refrigerated
delbrueckii
or frozen storage. The viability of probiotic organisms
Time pH Acidity Sugar CFU/mL is dependent on the level of oxygen in products, oxygen
(h) (% lactic acid) (mg/mL)
permeation of the package, fermentation time, and stor-
0 5.8 ± 0.0a 0.12 ± 0.0a 35.08 ± 0.09a 4.3 ± 1.76 · 105a age temperature (Shah, 2000). The viability of probiotic
24 4.5 ± 0.2b 0.30 ± 0.04b 36.32 ± 3.30a 7.6 ± 2.35 · 108b
48 3.7 ± 0.0c 0.74 ± 0.03c 29.62 ± 2.17b 15.4 ± 4.28 · 108c
bacteria is also affected by inhibitory substances such
72 3.6 ± 0.0c 0.95 ± 0.01d 28.45 ± 0.84b 11.0 ± 1.50 · 108ab as lactic acid produced during production and cold
Means and standard deviations for n = 3. The experimental values
storage. Other factors for loss of viability of probiotic
within rows that have no common superscript are significantly different organisms have been attributed to the decrease in pH
(p < 0.05) according to DuncanÕs multiple test range. of the medium and accumulation of organic acid as a
result of growth and fermentation (Hood and Zottola,
1988; Shah and Jelen, 1990). In this study, we found
produced only 0.74% titratable acidity expressed as lac-
both L. plantarum and L. delbrueckii could survive the
tic acid. It is probable that L. casei requires some essen-
high acidity and low pH in the fermented cabbage juice.
tial growth nutrients which are deficient in cabbage juice
(Pederson and Albury, 1969). Earlier studies have
reported that an antibacterial substance is present in
cabbage (Pederson and Fisher, 1944; Dickerman 4. Conclusion
and Liberman, 1952; Kyung and Fleming, 1994a). The
growth inhibitory substance of fresh cabbage was sug- Three lactic acid bacteria, L. casei, L. plantarum, and
gested to be carbohydrate in nature and of a low molec- L. delbrueckii were examined for their ability to utilize
ular weight (Dickerman and Liberman, 1952). Kyung cabbage juice for cell synthesis and lactic acid production
and Fleming (1994b) reported that fresh juice of Cecile without nutrient supplement. These lactic cultures grew
cultivar cabbage was inhibitory to the growth of lactic well in cabbage juice at 30 C, and the viable cell counts
acid bacteria, and the inhibition was eliminated when reached nearly 10 · 108 CFU/mL after 48 h of fermenta-
the cabbage was heated (steamed 10 min) before juice tion at 30 C. Both L. plantarum and L. delbrueckii were
extraction. capable of surviving the low pH and high acidic condi-
The data in Table 4 illustrate the effect of cold storage tions in fermented cabbage juice during cold storage at
on the viability of three species of lactic acid bacteria in 4 C. In contrast, L. casei could not survive the low pH
fermented cabbage juice. L. plantarum and L. delbrueckii and high acidity in fermented cabbage juice, and lost cell
were capable of surviving in the fermented cabbage juice viability completely after only 2 weeks of cold storage at
at 4 C for several weeks. For example, the viable cell 4 C. From the results of this study, it is concluded that
counts of L. plantarum and L. delbrueckii were still L. plantarum and L. delbrueckii could be used as probi-
4.1 · 107 and 4.5 · 105 mL 1, respectively, after 4 weeks otic cultures for production of a healthy beverage from
of storage at 4 C. However, L. casei was unable to sur- cabbage for vegetarians or consumers who are allergic
vive the low pH and high acidity conditions in fer- to lactose present in probiotic dairy products.
mented cabbage juice at 4 C and lost the cell viability
completely after only 2 weeks of cold storage. For the
maximum health benefits, the minimum number of pro- Acknowledgements
biotic organisms in a food product should be 106 CFU/g
Dr. Kyung Young Yoon received a post-doctoral fel-
lowship from Korea Science and Engineering Founda-
Table 4
Effect of cold storage on the viability of lactic cultures in fermented tion (KOSEF). This work was supported in part by
cabbage juice USDA Regional Project NE-1008.
Time CFU/mL
(weeks)
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