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Direct Spray Drying and Microencapsulation of Probiotic Lactobacillus Reuteri From Slurry Fermentation With Whey PDF
Direct Spray Drying and Microencapsulation of Probiotic Lactobacillus Reuteri From Slurry Fermentation With Whey PDF
ORIGINAL ARTICLE
Keywords
bacterial release, bacterial survival,
Lactobacillus reuteri, microencapsulation,
probiotic, spray drying, whey.
Correspondence
Christopher Beermann, Faculty of Food
Technology, Department of Biotechnology,
University of Applied Sciences Fulda, Marquardstrasse 35, Fulda, 36039, Germany.
E-mail: christopher.beermann@lt.hs-fulda.de
2013/0751: received 19 April 2013, revised
10 June 2013 and accepted 20 June 2013
doi:10.1111/jam.12293
Abstract
Aims: Formulations of dietary probiotics have to be robust against process
conditions and have to maintain a sufficient survival rate during gastric transit.
To increase efficiency of the encapsulation process and the viability of applied
bacteria, this study aimed at developing spray drying and encapsulation of
Lactobacillus reuteri with whey directly from slurry fermentation.
Methods and Results: Lactobacillus reuteri was cultivated in watery 20% (w/v)
whey solution with or without 05% (w/v) yeast extract supplementation in a
submerged slurry fermentation. Growth enhancement with supplement was
observed. Whey slurry containing c. 109 CFU g 1 bacteria was directly spraydried. Cell counts in achieved products decreased by 2 log cycles after drying
and 1 log cycle during 4 weeks of storage. Encapsulated bacteria were
distinctively released in intestinal milieu. Survival rate of encapsulated bacteria
was 32% higher compared with nonencapsulated ones exposed to artificial
digestive juice.
Conclusions: Probiotic L. reuteri proliferate in slurry fermentation with yeastsupplemented whey and enable a direct spray drying in whey. The resulting
microcapsules remain stable during storage and reveal adequate survival in
simulated gastric juices and a distinct release in intestinal juices.
Significance and Impact of the Study: Exploiting whey as a bacterial substrate
and encapsulation matrix within a coupled fermentation and spray-drying
process offers an efficient option for industrial production of vital probiotics.
Introduction
The Food and Agriculture Organization of the United
Nations and the World Health Organization underline in
their definition of probiotics that living micro-organisms
with health-promoting mechanisms have to be administered in adequate amounts to be effective (FAO and
WHO 2001). Therefore, dietary formulations of probiotics
have to be robust against industrial production processes
in a cost-effective way and should maintain a sufficient
survival rate after oral uptake and gastrointestinal tract
(GIT) transit.
Commonly accepted bacterial counts for dietary products are at least 106 CFU ml 1 (Kailaspathy and Chin
2000). A wide range of health benefits have been
described, such as immune response-modulating properties, improving gut barrier function and protecting effects
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slurry fermentation process coupled with direct encapsulation of bacteria by spray drying. At last, physical product characteristics and bacterial protective properties
against GIT conditions were investigated in vitro.
Materials and methods
Bacterial strain
Probiotic Lactobacillus reuteri (DSM 20016) was obtained
from the German Collection of Microorganisms and Cell
Cultures Leibnitz Institute DSMZ and cultured aerobically in 9 ml de ManRogosaSharpe (MRS) broth (pH
62) at 37C for 24 h.
Growth of Lactobacillus reuteri in whey
To characterize the growth of L. reuteri in whey,
approximately 103 CFU ml 1 bacteria were inoculated in
a 200-ml-volume fermentation of watery 20% (w/v) whey
solution (pH 60) with or without 05% (w/v) yeast
extract as supplement and cultivated in a agitating flask
culture at 37C. Before inoculation, whey solution was
sterilized in 80C water bath for 20 min. Bacterial cell
count was measured after 0, 24, 48 and 72 h by direct
plate counting as described below.
Batch slurry fermentation of Lactobacillus reuteri in a
continuous stirred-tank reactor (CSTR)
The growth of L. reuteri was observed in a laboratory-scale
reactor (Biostat A, Sartorius Ltd., Melsungen, Germany).
Approximately 104 CFU ml 1 bacteria were inoculated in
1 l batch slurry fermentation of watery 20% (w/v) whey
solution with 05% (w/v) yeast extract as supplement,
tested with adjustment at pH 50 and cultivated at 37C
with agitation using one six-wing rotating disc at 200 rpm,
respectively. Samples of 5 ml were taken to determine cell
counts after 0, 24, 48 and 72 h by direct plate counting as
described below.
Microencapsulation of Lactobacillus reuteri in whey
For the spray-drying process, a 48-h-incubated 200-ml
slurry culture of L. reuteri with at least 108 CFU ml 1 was
applied. The fermented slurry was spray-dried by a laboratory-scale spray dryer (B
uchi mini spray dryer B190; Flawil,
Switzerland) with two different outlet temperatures maintained at 55 2 and 65 2C and a flow rate of 500 Nl
h 1. Outlet temperatures were reached by adjusting different inlet temperatures (89 1 and 100 1C) and feed
levels from 2 to 4 ml min 1. Spray-dried powder samples
were collected from the cyclone and mixed gently.
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Results
1010
CFU g1
108
106
104
102
100
Before
After
Process
7 days
28 days
Storage
Figure. 1 Cell counts of Lactobacillus reuteri before and after spraydrying process with (
) 55C and (
) 65C outlet temperatures
and during storage for 7 and 28 days at 4C storage temperature.
The data shown represent the mean of at least three measurements.
Moisture (%)
55
65
61 08
68 03
55 03
49 03
Lactobacillus reuteri was spray-dried at 55 and 65C temperatures using the whole slurry fermentation with whey
directly as feed. Loss of viable bacteria after spray drying
is illustrated in Fig. 1. For all experiments, whey solutions
with 16 ( 15) x 109 CFU g 1 bacteria were applied to
the encapsulation process. Dried product contained 25
( 17) x 107 CFU g 1 bacteria directly after processing
independently from used outlet temperatures.
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100
80
60
40
20
0
0
05
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Journal of Applied Microbiology 115, 1029--1036 2013 The Society for Applied Microbiology