The document discusses microencapsulation of probiotics for industrial applications. It describes how various biopolymers like gelatin, alginate, chitosan can be used to encapsulate probiotics and protect them from harsh environments in the GI tract. Spherical polymer beads produced using extrusion or emulsification techniques encapsulate probiotic biomass. Chitosan in particular is a promising encapsulation material as it is non-toxic, biodegradable and can be ionically cross-linked for stability. Alginate encapsulation using calcium chloride has effectively protected L. acidophilus from gastric acid conditions. The encapsulation techniques aim to enhance probiotic viability during food processing and consumption.

1. Submitted by:
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2. objective ;
 determine the role of pH in forming
thermally stable whey protein isolate (WPI)
soluble aggregates (SA).
 evaluate the functionality of SA formed
under a range of conditions.
 evaluate SA stability in a model beverage.

3.  Proteins are important ingredients in a
variety of beverages including those designed
for recovery after exercise.
 Protein-containing beverages can be
generally sub-divided based on pH and
thermal processing.

4. Composition:
Whey protein is a mixture of the following:
 Beta-lactoglobulin
 Alpha-lactalbumin
 Bovine serum albumin
 Immunoglobins.
 A whey protein may in the form of:

5.  Whey protein concentrate - WPC contains low
levels of fat and CHO. Lower end concentrates
tend to have 30% protein and higher end up to
90%
 Whey protein isolate - WPIs are further
processed to remove all the fat and lactose. WPI
is usually at least 90% protein
 Whey protein hydrolysate - WPH is considered
to be the "predigested" form of whey protein as
it has already undergone partial hydrolysis - a
process necessary for the body to absorb
protein. WPH doesn't require as much digestion
as the other two forms of whey protein.

6.  protein stock solutions (15% w/w) were
prepared by slowly dissolving the protein
powder in ~90% of the total buffer (5 mM
sodium phosphate, pH 6.8, with 0.02%
sodiumazide.
 Protein solutions were stirred at room
temperature (23 C) and left overnight for
complete hydration.

7.  Whey protein solutions prepared by
dispersing WPI powders will be called
“native” to distinguish them from soluble
aggregate solutions.
 Aggregation: Any abnormal association of
misfolded proteins (or parts of proteins).
Aggregation is a process that begins with the
abnormal association of as few as two
molecules and that has the potential to form
larger structures that are visible by
microscopy.

8.  Stock solutions were diluted to 5 or 7% w/w
protein in buffer (5mMsodium phosphate, pH
6.8, with 0.02% sodium azide was the buffer
for this and all subsequent experiments) and
adjusted to pH6.5 or 7.5.
 measuring the optical density at 400 and 600
nm using aShimadzu UV-160U
spectrophotometer.

9.  Thermal stability was evaluated by
determining turbidity as fallow,
 viscosity,
 solubility,
 aggregate size

10.  SA improved thermal stability of two sources
of WPI.
 whey protein SA can be improve the storage
stability of beverages by reducing turbidity
and decreasing age-associated increases in
protein aggregation and viscosity.

11.  Alizadeh-Pasdar, N., & Li-Chan, E. C. Y. (2000). Comparison of protein surface
hydrophobicity
 measured at various pH values using three different fluorescent
 probes. Journal of Agricultural and Food Chemistry, 48(2), 328e334.
 Beecher, J. W., Drake, M. A., Luck, P. J., & Foegeding, E. A. (2008). Factors
regulating
 astringency of whey protein beverages. Journal of Dairy Science, 91(7),
 2553e2560.
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 with special consideration of cold-set gels derived from heat-denatured whey.
 Trends in Food Science & Technology, 9(4), 143e151.
 Code of Federal Regulations. Title 21, sec. 114. (2001a). Acidified foods.
http://www.
 access.gpo.gov/nara/cfr/waisidx_01/21cfr114_01.html Accessed 29.09.11.
 Code of Federal Regulations. Title 21, sec. 113. (2001b). Thermally processed low-
acid
 foods packaged in hermetically sealed containers. http://www.access.gpo.gov/
 nara/cfr/waisidx_01/21cfr114_01.html Accessed 29.09.11.
 Donato, L., Schmitt, C., Bovetto, L., & Rouvet, M. (2009). Mechanism of formation
of
 stable heat-induced b-lactoglobulin microgels. International Dairy Journal, 19

12.  Some industries used natural source to made
carbonated soft drinks (malt drinks from
barley malt or wheat or maize)
 Haze problem (degree of transparency)
 Haze due to proteins (protease)
 Complex proteins and polyphones react.
 Hot break
 Chill break
 Chemicals (diversion f and lucilite)

14.  Fruit juices have been traditionally
concentrated by multi-stage vacuum
evaporation, resulting in a loss of fresh juice
flavors,color degradation and a ‘‘cooked’’
taste due to the thermal effects.
 New membrane processes, including
membrane distillation and osmotic
distillation, and integrated membrane
processes are still being identified and
developed in concentrated fruit juice
processing to improve product quality and
reduce energy consumption.

15.  Advantages of RO over traditional
evaporation.
 low thermal damage to product.
 reduction in energy consumption.
 lower capital investments.

16.  Direct osmosis concentration (DOC) is
another membrane process capable of
concentrating fruit juice at low temperatures
and low pressures, thereby maintaining
original flavor and color characteristics of
fruited juices.
 The principle uses an osmotic agent (OA)
solution to establish an osmotic pressure
gradient across a semi permeable membrane
and thus remove water from a single
strength.

17.  Membrane distillation is a relatively new
membrane process in which two aqueous
solutions, at different temperatures, are
separated by a micro porous hydrophobic
membrane.
 In these conditions a net pure water flux
from the warm side to the cold side occurs.

18.  The driving force is the vapour pressure
difference between the two solution–
membrane interfaces due to the existing
temperature gradient.
 The phenomenon can be described as a three
phase sequence.
 formation of a vapour gap at the warm
solution–membrane interface.

19.  transport of the vapour phase through the
microporous system.
 its condensation at the cold side membrane–
solution.

20.  Ali, F., Dornier, M., Duquenoy, A., & Reynes, M. (2002). Transfer of
 volatiles through PTFE membrane during osmotic distillation. In
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 membrane processes, Toulouse, France.
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 flux prediction in apple juice concentration by reverse osmosis.
 Journal of Membrane Science, 127(1), 25–34.
 Alvarez, S., Riera, F. A., Alvarez, R., & Coca, J. (1998). Permeation of
 apple aroma compounds in reverse osmosis. Separation Purification
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 S. N., & Panyor, L. (2000). A new integrated membrane process for
 producing clarified apple juice and apple juice aroma concentrate.
 Journal of Food Engineering, 46(2), 109–125.
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 flux and aroma compounds rejection in a reverse osmosis concentration
 of apple juice model solutions. Industrial Engineering
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 Bailey, A. F. G., Barbe, A. M., Hogan, P. A., Johnson, R. A., &

21. Recent advances in
microencapsulation of probiotics
for industrial applications

22.  Because of their perceived health benefits,
probiotics have been incorporated into a
range of dairy products, including yoghurts,
soft-, semi-hard and hard cheeses, ice
cream, milk powders and frozen dairy
desserts.
 This review focuses mainly on current
knowledge and techniques used in the
microencapsulation of probiotic
microorganisms to enhance their viability
during fermentation, processing and
utilization in commercial products.

23.  Gelatin is useful as a thermally reversible
gelling agent for encapsulation.
 encapsulated Bifidobacterium cells in a
mixed gel composed of alginate, pectin and
whey proteins.
 Because of its amphoteric nature, it is also
an excellent candidate for incorporating with
anionic-gelforming polysaccharides, such as
gellan gum.

24.  These hydrocolloids are miscible at because
they both carry net negative charges and
repel one another.
 net charge of gelatin becomes positive when
the pH is adjusted below its isoelectric point
and causes a strong interaction with the
negatively charged gellan gum.

25.  Encapsulation of probiotics in a
biodegradable polymer matrix has a number
of advantages.
 Various polymer systems have been used to
encapsulate probiotic microorganisms to
protect against low pH and high bile
concentrations and to enhance physical
stability during downstream processing.

26.  Spherical polymer beads with diameters
ranging from 0.3 to 3.0 mm and immobilizing
active biomass are producedusing extrusion
or emulsification techniques.

27.  The biopolymer chitosan, the N-deacetylated
product of the polysaccharide chitin, is
gaining importance in the food and
pharmaceutical field because of its unique
polymeric cationic character, good
biocompatibility, non-toxicity and
biodegradability.

28.  Chitosan can be isolated from crustacean
shells, insect cuticles and the membranes of
fungi.
 Chitin is a polymer of b-(1-4)-2-acetamido-2-
deoxy-D-glucopyranose.
 In order to achieve sufficient stability,
chitosan gel beads and microspheres can be
ionically cross-linked with polyphosphates.

29.  Alginic acid, a natural polymer, is a
polyuronic acid that is extracted from
seaweeds.
 It is composed of various proportions of 1e4
linked b-D-mannuronic (M) and a-Lguluronic
(G) acid.
 Addition of sodium alginate solution to a
calcium solution, interfacial polymerization
is instantaneous, with precipitation of
calcium alginate followed by a more gradual
gelation of the interior as calcium ions
permeate thhrough algenate system.

30.  The encapsulation method, with sodium
alginate in calcium chloride (CaCl2), has
been used to encapsulate L. acidophilus to
protect this organism from the harsh acidic
conditions in gastric fluid.

31.  Anal, A. K., Bhopatkar, D., Tokura, S., Tamura, H., & Stevens, W. F.
 (2003). Chitosan-alginate multilayer beads for gastric passage and
 controlled intestinal release of protein. Drug Development and
 Industrial Pharmacy, 29, 713e724.
 Anal, A. K., & Stevens, W. F. (2005). Chitosan-alginate multilayer
 beads for controlled release of ampicillin. International Journal of
 Pharmaceutics, 290, 45e54.
 Anal, A. K., Stevens, W. F., & Remun˜a´n-Lo´pez, C. (2006). Ionotropic
 cross-linked chitosan microspheres for controlled release of
 ampicillin. International Journal of Pharmaceutics, 312, 166e173.
 Audet, P., Paquin, C., & Lacroix, C. (1988). Immobilized growing lactic
 acid bacteria with k-carrageenan-locust bean gum gel. Applied
 Microbiology and Biotechnology, 29, 11e18.
 Audet, P., Paquin, C., & Lacroix, C. (1990). Batch fermentations with
 a mixed culture of lactic acid bacteria immobilized separately in
 k-carrageenan locust bean gum gel beads. Applied Microbiology
 and Biotechnology, 32, 662e668.
 Audet, P., Paquin, C., & Lacroix, C. (1991). Effect of medium and
 temperature of storage on viability of LAB immobilized in
 k-carrageenan-locust bean gum gel beads. Biotechnology
 Techniques, 5, 307e312.

32. The application of NMR methods for
detection of adulteration of wine,
fruit juices, and olive oil.

33. Nuclear magnetic resonance NMR spectroscopy
is rapidly gaining importance in fruit juice
quality control.
NMR allows analysis in two ways within one
experiment,
 Targated
 un targeted

34.  Targeted stands for the safe identification
and consequent quantification of individual
compounds.
 whereas untargeted means the detection of
all deviations visiblebyNMRusing statistical
analysis based on normality models.

35.  NMR has been perceived as a tool for
structure verification, elucidation and purity
analysis.
 NMR spectrometer with especially shielded
magnet technology, which in the case of a
400-MHz system ensures that the five gauss
line of the magnet stays within the magnet
can.

36.  The system most efficiently operates with
flow-injection NMR having a 4-mm o.d. flow
probe using water as transfer medium.
 A liquid handler is used for sample
preparation, intermediate storage and
transfer.
 Preparation consists of buffer addition and
consequent mixing to adjust to a pH.

37.  To minimize handling errors, the samples are
provided in barcoded cryovials.
 For each juice a multitude of parameters
related to quality and authenticity are
evaluated simultaneously from a single data
set acquired within a few minutes.

39.  The targeted approach provides the
identification and quantification of individual
compounds.
 Here, NMR spectroscopy provides a clear
advantage over classical analysis techniques
as it allows the identification and
quantification of many compounds in a
mixture simultaneously

40.  In addition to compound quantification, an
exhaustive statistical analysis is applied to
the same data.
 As a first step, the type of fruit is estimated.
 With conventional analysis the addition of
mandarin juice to orange juice is difficult to
detect, but with our NMR methods and
models we can detect mandarin juice at a
level.

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