Nanoprobiotics - An Innovative Trend in Probiotic World

e-ISSN: 2582-5208
International Research Journal of Modernization in Engineering Technology and Science

( Peer-Reviewed, Open Access, Fully Refereed International Journal )
Volume:03/Issue:11/November-2021 Impact Factor- 6.752 www.irjmets.com
NANOPROBIOTICS - AN INNOVATIVE TREND IN PROBIOTIC WORLD
Bhargavi B*1, Ramachandra B*2, Pushpa B.P*3, Prabha R*4
*1M. Tech Scholar, Dept. of Dairy Microbiology, DSC, KVAFSU, Bengaluru, Karnataka, India.
*2Assistant Professor, Dept. of Dairy Microbiology, DSC, KVAFSU, Bengaluru, Karnataka, India.
*3Assistant Professor, Dept. of Dairy Chemistry, DSC, KVAFSU, Bengaluru, Karnataka, India.
*4Associate Professor and Head, Dept. of Dairy Microbiology, DSC, KVAFSU, Bengaluru,
Karnataka, India.
ABSTRACT
Microorganisms are extremely small unicellular living things that are visible under microscope. Microbes are
present everywhere including human body, some are beneficial termed probiotics while some are harmful
called pathogens involved in causing various diseases. Human gastrointestinal (GI) tract has probiotics that
extend therapeutic benefits. Nanoparticles with probiotics are called nanoprobiotics. Nanoprobiotics is a field
that focuses on the application of nanoscience into the probiotic related world. Microorganisms used as
probiotics are Lactobacillus spp, Bifidobacterium spp, Saccharomyces boulardii. Encapsulation has become an
effective way to improve survival of probiotic organisms in GI tract by acting as a protective transporter for
deliverance at a targeted level. The available technologies employed to encapsulate probiotic cells are micro
and nanoencapsulation techniques which use sodium alginate, polyvinyl alcohol, pectin etc., as encapsulating
material. Nanoencapsulation is a process of entrapping of active ingredients in nanometer (10-1000 nm) sized
capsules. It is a technique which appears to be a promising alternative approach, has a potential to improve
bioavailability, enhances controlled release and enables an accurate targeting of the probiotics in a greater
amount than microencapsulation techniques, because of the large surface area and enhanced preservation
against heat treatment. Electrospinning is a well-known nanoencapsulation technique, which is efficient and
cost-effective for fabrication of fiber mats in nanosize for encapsulation of probiotics.
Keywords: Nanoprobiotics, GI Tract, Encapsulation, Nanoencapsulation, Electrospinning, Nanofibers.
I. INTRODUCTION
Microorganisms are extremely small unicellular living things that are visible under microscope such as bacteria,
archea, yeast, mold, protozoa, algae. They are ubiquitous in nature present in human body. They are of
commensals, beneficial and pathogens. Foods plays an important role in maintaining the balance of human GI
tract. Food components prebiotics act as food for good bacteria termed as probiotics. Most of the probiotics
belongs to LAB such as. Lactobacillus sp, Lactococcus sp, Streptococcus thermophilus etc. It has been
recommended that food containing probiotic bacteria should be in the range of 10 8 -109 cfu/g right before
ingestion to ensure that sufficient therapeutic minimum of 106 -107 cfu/g could reach the colon [1]. Antibiotic
therapy kills the good bacteria hence balancing of probiotics in human body is necessary. When probiotics are
taken orally reaches gastro-intestinal tract but survive in low numbers because of acid and bile conditions in GI
tract. Necessity is the encapsulation of probiotics using the nanoparticles termed as nanoprobiotics.
Nanotechnology is a fast-rising industry which exploits the benefits in nano scale dimensions and
characteristics for application in the macro world. Probiotics are defined as “live microorganisms that, when
administered in adequate amounts, confer a health benefit on the host.” [2]. The size of nanoprobiotics ranges
from 0.5– 0.8 to 2 - 9µ. Nanofibers are produced by the electrospinning process whose diameter is less than
1000 nm used as encapsulating material. Nanoprobiotics is a field that focuses on the application of
nanoscience into the probiotic related world which increases the viability and releases the probiotics at target
sites [3].
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II. LITERATURE REVIEW
Microorganisms Used as Probiotics
Bifidobacterium Non lactic acid
Lactic acid producing
species producing Yeast References
bacteria
(LA +AA) bacteria
Bifidobacterium Escherichia coli
Lactobacillus acidophilus
adolescentis Nissle
Lactobacillus casei Bifidobacterium animalis
Lactobacillus reuteri Bifidobacterium bifidum
Lactobacillus johnsonii Bifidobacterium longum
Saccharomyces
Lactobacillus crispatus Bifidobacterium breve
Propionibacterium boulardii
Lactobacillus rhamnosus freudenreichii
Bifidobacterium infantis
GG [4]
Enterococcus faecalis
Enterococcus faecium Bifidobacterium lactis
Bacillus coagulans
Characteristics & Therapeutic Benefits of Probiotics
Probiotic Strain Characteristics: Human origin, Acid and bile stability, Adherence to human intestinal cells,
Persistence in human intestinal tract, Production of antimicrobial substances, Antagonism against pathogenic
bacteria, Safety in food, clinically validated and documented health effects
Probiotics Therapeutic benefits: Control of intestinal pathogens, Help in lactose use in Lactose
intolerants, Boost up the host immune response, Reduction in serum cholesterol concentration,
Anticarcinogenic activity [5].
Definition of Nanofibers
An emerging technology is the production of nanofiber. The word “nano” comes from Greek and means “dwarf”.
A nanometer is a thousandth of a thousandth of a thousandth of a meter (10-9 m). One nanometer is about
60,000 times smaller than a human hair in diameter, or the size of a virus. These fibers have diameters of less
than 1000 nm, produced by the electrospinning process. Different materials such as inulin, pectin, alginate are
used to make nanofibers which improves the viability of probiotics [6].
Encapsulation of Probiotics
Encapsulation is a “process by which probiotic is coated with, or entrapped within, food grade viscous
material”. Encapsulation protects and maintains the viability and efficacy of probiotics in the food products and
GI tract, promotes the controlled release and optimize the delivery at site of action and prevent probiotics from
multiplying in food otherwise causes change in sensory characteristics. Protective shell of encapsulate should
be of food grade, GRAS status, Biodegradable and should form barrier between internal phase and
surroundings [7].
Materials used for Encapsulation
Materials Examples References
Plant Based
Starch and their
Amylose, amylopectin, dextrins, maltodextrins, cellulose and
derivatives
gluten
Plant exudates and Gum Arabic, galactomannans, pectins and soluble soybean
extracts polysaccharides
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Marine Based [1]
Marine extracts Chitosan (crab - Callinectes sapidus)

Microbial Based
Dextran (Leuconostoc spp.), xanthan (Xanthomonas campestris)
Bacteria
and gellan (Spingomonas spp.)
Yeast Chitosan (Saccharomyces cerevisiae)
Carrageenan (Chondrus crispus – Red alga) and alginate
Algae
(Macrocystis pyrifera - Brown alga)
Animal Based
Proteins Casein, whey proteins and gelatin
Fatty acids and fatty alcohols, waxes (beeswax), glycerides and
Lipid materials
phospholipids
Synthetic materials
Other Synthetic materials PVA, PVP, Paraffin (GRAS)
Microencapsulation
Microencapsulation is the process of applying a shell to sensitive probiotic to protect them from their external
environment and the probiotic cells are retained within an encapsulating matrix. It is a “process in which the
probiotic cells are incorporated into an encapsulating matrix or membrane that can protect the cells from
degradation by the damaging factors in the environment and release at controlled rates under particular
conditions.” It protects the probiotics in food processing from high temperatures, oxygen stress and pH
changes. After ingestion, probiotics encounter various types of harsh environmental conditions in Stomach
(acid) and in small intestine (bile &enzymes). Probiotics size ranges from 1 to 10 μm, Microgels used for
encapsulation of probiotics whose diameters ranges from 1 to 1000 μm [8].
III. METHODS OF MICROENCAPSULATION
The encapsulation techniques applied to probiotics includes emulsion, extrusion and spray drying [9].
Emulsion: Polymers used for coating oil-in-water emulsion droplets along with core material and
homogenized well before chemically hardened to form a shell. Bead size with a range of 25-35 μm.
Extrusion: Dispersion of active material in a molten carrier solution and forcing of the mixture into a
dehydrating liquid. Bead size ranges from 0.3-3mm depends on distance between syringe and hardening
solution (cacl2).
Spray drying: Automize the infeed dispersion of the active and core mix and further dehydration of the
automized particles. Inlet temperature – (126-154°C) and Outlet temperature – (73-87°C). Particle size is less
than 40 μm.
Encapsulated Lactobacillus acidophilus and Bifidobacterium bifidum by Emulsion & Extrusion
Techniques
Lactobacillus acidophilus and Bifidobacterium bifidum are encapsulated in sodium alginate by emulsion and
extrusion techniques. The viability of microencapsulated Lactobacillus acidophilus using emulsion technique at
10°C for 90 days shown the final count of 9.20 log10 cells /g for which initial count was 9.50 log10 cells /g where
as for Bifidobacterium bifidum was 9.21 log10 cells /g for which initial count was 9.45 log10 cells /g and
microencapsulated Lactobacillus acidophilus using extrusion technique at 10°C for 90 days shown the final
count of 7.17 log10 cells /g for which initial count was 7.32 log10 cells /g where as for Bifidobacterium bifidum
was 7.11 log10 cells /g for which initial count was 7.25 log10 cells /g [10].

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Microencapsulation vs Nanoencapsulation of Probiotics
Microencapsulation Nanoencapsulation References
Particle size - Microns Particle size – Nanometer
Lower surface area Higher surface area
Increased shelf life Enhanced shelf life
Reach targets Precised targeting
Probiotics
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Nanofiber
IV. NANOENCAPSULATION OF PROBIOTICS
Nanoencapsulation is a “Process of entrapping of probiotics in nanometer (10-1000nm) sized particles”.
Nanoencapsulation is a technique which appears to be a promising alternative approach, has a potential to
improve bioavailability, enhances controlled release, and enables an accurate targeting of the live organisms in
a greater amount than microencapsulation techniques. In addition, nanoencapsulation brings forward some
benefits such as increased immobilization efficacy because of the large surface area and enhanced preservation
against heat treatment as a result of decreased thermal conductivity with volume. There are two methods of
nanoencapsulation of probiotics - Electrospraying and Electrospinning [12].
V. ELECTROSPRAYING & ELECTROSPINNING
Principle of Electrospraying & Electrospinning: Electrospraying and Electrospinning methods works under
the same principle of Electrohydrodynamic processes. Both methods utilize electrically charged jet of polymer
solution for production of fibers or particles in micron, submicron and nanoscale. These methods are facile, cost
effective and flexible [13].
Methods of Nanoencapsulation of Probiotics
Electrospraying: Electrospraying is a process, in which polymer liquids in small capillary needles as subjected
to sufficiently high voltage, the polymer liquid at the mouth of the small needles is charged then a columbic
repulsion force is generated, then the polymer liquid is deformed to form Taylor cone. When the electrostatic
repulsion overcomes the surface tension of Taylor cone, fine uniform liquid droplets are atomized and ejected
from the conetip. Electrospraying Forms automized droplets used for encapsulating of probiotic cultures by
protein or polysaccharides as entrapping materials [14].
Electrospinning: Electrospinning is a nanofiber fabrication technique that applies high voltage to draw a
polymer solution from a spinneret tip to grounded collector. Upon charging, a jet of polymer solution emerges

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from the needle tip, forming a cone known as Taylor cone, which breaks out into jet, when the critical voltage is
achieved. The electrical between the tip and grounded collector draws the jet in a whipping motion and
stretches it into a continuous fiber of nanosized diameter as a non-woven mat on the grounded collector.
Generally, for electrospinning flow rates (0.4 to 1.6 ml/h), voltage values (15 to 27 kV) and distances between
flat collector and Taylor cone (6 to 15 cm) are maintained. Electrically charged jet of polymer solutions (Ex:
chitosan, sodium alginate) forms a nanoscale fiber of particles [14].
Electrospinning Vs Electrospraying
Process and equipment are same for both electrospinning & Electrospraying
 Higher viscosity Solutions are used to produce Fibers
 Lower Viscosity Solutions are used to produce Particles or Powders
 Attachment of probiotics in both the processes depends upon electrostatic force, negative charge of bacteria
and viscous nature of material used [15].
Fibers and Particles - Possible Diameter is 50nm to 1μm
Fibers Particles
Preparation of Nano Lactobacillus acidophilus
Lb. acidophilus grown in sterile MRS broth (incubation 370C/24 h) collected by centrifugation at 10000 rpm for
15 min at 4 °C, washed twice and resuspended in PBS (pH 7.4). Okara and PVA stirred at 100 °C using a
magnetic stirrer (600 rpm), until complete dissolution. Okara (soluble fraction of Soya pulp) 7%, PVA 8%, 86%
water and Lb. acidophilus 2% was taken and subjected to electrospinning (25 °C). The prepared spinning
solutions were inserted in a 1 ml syringe with needle of 0.5 mm diameter, used flow rate of 0.2–0.5ml/h,
voltage of1.1kV and distance between electrode tip & collector disc is 12 cm at room temperature with
humidity of 50%. The nanofibers were gathered by a rotating mandrel collector then vacuum Packed and
stored at 4°C for later usage [16].
Results And Findings
SEM (Scanned electron microscopy) Image
Spun nanofiber obtained has diameter of 465 nm with 7 % moisture

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In vitro viability of strain after encapsulation process
 74 % Viability rate noticed after 21 days when stored at 5 °C
 Initial count was 9.58 log cfu/g after electrospinning the final count observed was 7.13 log cfu/g which
shows a slight decrease in the probiotic viability [16].
Nanoencapsulation of Bifidobacterium sp
A. Preparation of polymer solutions
All the solutions and the glassware were autoclaved at 121 °C for 15 min to achieve sterility. CS solution (1
wt%) was provided by adding an appropriate amount of its powder into acetic acid (0.5 M) over a period of 5-6
hours on a magnetic stirrer (600 rpm). PVA solution (15 wt%) was prepared by dissolving an adequate amount
of PVA powder in sterile deionized water, at 90 ºC, under stirring at 3 h. Then, the mixture CS/PVA (10/90,
w/w) was obtained by blending polymers and stirring the polymer solution for 6 h to acquire a uniform blend
[3].
B. Preparation of bifidobacterial cell suspensions
Lyophilized cells of B. animalis subsp. lactis Bb12 was grown in sterile MRS broth and agar medium and
incubated under anaerobic conditions for 24 hrs at 37 °C. Then, the cells were collected by centrifugation (6000
rpm, 20 min), washed twice with PBS (pH 7.4), and re-suspended in skimmed milk to nearly 3 × 109 cell/ml [3].
C. Electrospinning of bacterial suspensions
The prepared spinning solutions were inserted in a 10 mL syringe fitted with a 23-gauge steel needle. The
electrospinning process used a flow rate of 0.1 ml/h, a positive high voltage of 18 kV, and tip-to-collector
distance (TCD) of 15 cm. The nanofibers were gathered by a rotating mandrel collector, which was covered
with an aluminum foil [3].
Results and Findings
SEM (Scanned electron microscopy) Images
a. b.
Average Diameter of Nanofibers a. 139.06 ± 75.02nm b. 217.56 ± 62.74nm

The diameter of strain loaded nanofiber is greater than the free nanofiber, the diameter increased to around
217 nm, revealing that the strain could be successfully encapsulated within the nanofiber mats [3].
State of strain Viability rate (%)
Free strain (log cfu/g) Encapsulated strain (log cfu/g)
Viability 9.47 8.40 88.70
The resistance of the strain to electrospinning process during encapsulation was revealed by in vitro viability
tests. The viability of the Bifidobacterium sp cells was established after encapsulation. Above table shows that
the viability rate after the electrospinning process was 88.70%, which shows a slight decrease in the probiotic
viability [3].
Preparation of Nano Lactobacillus paracasei KS-199
Optical micrographs of free strain and encapsulated strains

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(A) free and (B) encapsulated strain (strain-loaded nanofiber mats)

Fig.(A) optical micrographs of free strain, clearly showing the rod-shape morphology of the strain.
Fig.(B)shows the morphology of the encapsulated strains, in other words, that of the strain-loaded
nanofiber mats [12].
SEM Image of Blank & Strain Loaded Nanofiber Mats
Average Diameter of Nanofibers A. 305 ± 29nm (blank nanofiber) B.842 ± 72nm (strain-loaded nanofiber)
Figures shows the SEM images of blank and strain-loaded nanofiber mats along with their average diameter
distributions. It shows that the nanofibers mats fabricated from PVA/SA mix had uniform, well-defined, smooth
and bead-free structure. Majority of fiber mats were in the size of between 170–360 nm with an average size of
305 nm in diameter. On the other hand, the incorporation of L. paracasei into the spin solutions yielded in
beaded fiber structure (Fig. B). The diameter increased to around 842 nm, revealing that the strain could be
successfully encapsulated within the nanofiber mats [12].
In-vitro viability of Probiotic Lactobacillus paracasei KS-199 after Nanoencapsulation Process.
State of strain Viability rate (%)
Free strain (log cfu/g) Encapsulated strain (log cfu/g)
Viability
9.98 ± 0.85 8.57±0.69 85.87±0.78
The resistance of the strain to electrospinning process during encapsulation was revealed by in vitro viability
tests based on plate counting technique. For this purpose, the viability of the L. paracasei cells was established
after encapsulation. Above table shows that the viability rate after the electrospinning process was 85.87%,
which shows a slight decrease in the probiotic viability [12].
Viability of free and nanoencapsulated strains in simulated gastrointestinal conditions
State of strain
Encapsulated strain
Free strain
After
Before Before After Redution
Exposure Reduction Viability
exposure Viability Exposure Exposure In
To in viability rate
to rate (%) to GJ/BS to GJ/BS viability
GJ/BS (log cfu/g) (%)
Viability GJ/BS (log (log (log cfu/g)
(log
(log cfu/g) cfu/g) cfu/g)
cfu/g)

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70.8±2.
9.79±0.6 6.28±0.4 64.1±17 3.51±0.3 8.51±0.7 6.02±0.36 2.49±0.3
5
The viability rate of encapsulated strain is 70.8% greater than free strain which is 64.1%, clearly showing that
alginate-based nanoencapsulation considerably improved the survival percentage of L. paracasei KS-199
following exposure to the simulated gastric juice. There was a reduction in viabilities of 3.51 and 2.49 log cfu/g
for the free and encapsulated cells, respectively after the exposure of gastric digestion, which demonstrates that
it was possible to increase the viability of the cells by around 1 log [12].
Synbiotic (prebiotic + probiotic) nanoencapsulation [Inulin + Lactobacillus fermentum]
PVA solution (20% -w/v), Sodium alginate (2% - w/v), Inulin (15% -POS) and Lb. fermentum was dissolved in
sterile distilled water at 6:1:2:1 ratio and subjected to electrospinning process (25°C) The prepared spinning
solutions was fed at the rate of 1.5 ml/h at a tip-to-target distance of 10 cm, Voltage of 20 kV and ~50% RH.
The electrospun nanofiber mats were collected from the aluminum foil [17].
SEM (Scanned electron microscopy) Image
Entrapment of bacteria increased fibre size up to 400 nm with encapsulation efficacy of 78 %

Count (logcfu/g)
Conditions
Conditions
Initial count 9.00
Survivability (GI in-vitro)
Free cells 3.78
Nanoencapsulated 7.90
Storage Temp. (0C) Free cells Nanoencapsulated
4 2.80 8.00
-18 2.10 7.60
The nanoencapsulated strain has showed the higher viability of 7.90 in GI whereas free cells showed the lesser
viability of 3.78 because of high acid and bile conditions, The nanoencapsulated strain stored at 40C has showed
more viability of 8.00 than free cells i.e., 2.80 and also at -18 0C nanoencapsulated strain has higher viability of
7.60 than free cells which was 2.10 [17].
Release of Probiotics from Nanofiber after Ingestion

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When probiotic encapsulated nanofiber taken orally, reaches stomach, encapsulated probiotic travels safely
past the stomach i.e., nanofiber protects the probiotic from acid conditions (pH-1.8-5.0) and is released in the
small intestine (pH – 6.5-7.5). The probiotic growth and bacterial multiplication take place throughout the
intestinal tract. The degraded nanofiber (encapsulating material) acts as prebiotic in colon [11].
VI. CONCLUSION
Probiotics provides the therapeutic benefits, But in food processing because of harsh conditions such as heat
treatment and storage temperatures, decreases the viability of probiotics and also in GI tract because of acid &
bile conditions, affects the viability of probiotics and stops them from reaching to target site therefore action on
probiotic is needed. Best method is the nanoencapsulation which protects and helps in release of probiotics at
targeted site. Selection of good encapsulating material & techniques are optimized. Proof of high viability nano
encapsulated probiotics was observed. Nanotechnology techniques have improved the viability and
survivability in all the probiotics which makes nanoencapsulation a thrust area.
VII. REFERENCES
[1] Nazzaro, F., Fratianni, F., Coppola, R., Sada, A. and Orlando, P., 2009. Fermentative ability of alginate-
prebiotic encapsulated Lactobacillus acidophilus and survival under simulated gastrointestinal
conditions. J Funct Foods., 1: 319.
[2] International Scientific Association for Probiotics, 2013.
[3] Mojaveri, S.J., Hosseini, S.F. and Gharsallaoui, A., 2020. Viability improvement of Bifidobacterium
animalis Bb12 by encapsulation in chitosan/poly(vinyl alcohol) hybrid electrospun fiber mats.
Carbohydrate Polymers. DOI: https://doi.org/10.1016/j.carbpol.2020.116278.
[4] Anusha, R.L., Umar, D., Basheer, B. and Baroudi, K., 2015. The magic of magic bugs in oral cavity:
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[5] Kerry, R.G., Patra, J.K., Gouda, S., Park, Y., Han-Seungshin. and Das, G., 2018. Benefaction of probiotics
for human health. A review. J. Food and Drug Analysis., 26(3): 927-939.
DOI: 10.1016/j.jfda.2018.01.002.
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[7] Digambar, K., Sujatha, K., Bruntha Devi, P. and Shetty, H.P., 2017. Recent developments on
encapsulation of lactic acid bacteria as potential starter culture in fermented foods. A review. J. Food
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[9] Liu, H., Gong, J., Chabot, D., Miller, S.S., Cui, S.W., Zhong, F. and Wang, Q., 2018.Improved survival of
Lactobacillus zeae LB1 in a spray dried alginate-protein matrix. Food Hydrocolloids., 78:100-108.
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[10] Ozer, B., Uzun, Y.S. and Kirmaci, A.V., 2008. Effect of Microencapsulation on Viability of Lactobacillus
acidophilus LA-5 and Bifidobacterium bifidum BB-12 during Kasar Cheese
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[11] Lingli, D. and Hui, Z., 2020. Recent Advances in Probiotics Encapsulation by Electrospinning. ES Food
Agrofor., 2: 3-12. DOI: https://dx.doi.org/10.30919/esfaf1120.
[12] Yilmaz, M.T., Edu, M.K., Taylana, O., Karakasb, Y. and Dertlib, E., 2020. An alternative way to
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gastrointestinal conditions and in kefir. DOI: https:// doi.org/ 10.1016/j.carbpol.2020.116447.
[13] Frenot, A. and Chronakis, I., 2003. Polymer nanofibers assembled by electrospinning. Curr Opin Colloid
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[14] Alonso, S., 2016. Novel Preservation Techniques for Microbial Cultures. Food Engineering Series. DOI:
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[15] Chakraborty, S., I -Chien Liao., Adler, A. and Leong K.L., 2009. Electrohydrodynamics: A facile
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[16] Fung, W.Y., Yuen, K.H. and Liong M.T., 2011. Agrowaste- Based Nanofibers as a Probiotic Encapsulant:
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[17] Duman, D. and Karadag, A., 2020. Inulin added electrospun composite nanofibres by electrospinning
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simulated gastrointestinal digestion. Int. J. Food Sci and Tech. DOI: 10.1111/ijfs.14744.

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e-ISSN: 2582-5208

International Research Journal of Modernization in Engineering Technology and Science

www.irjmets.com @International Research Journal of Modernization in Engineering, Technology and Science

Marine extracts Chitosan (crab - Callinectes sapidus)

www.irjmets.com @International Research Journal of Modernization in Engineering, Technology and Science

www.irjmets.com @International Research Journal of Modernization in Engineering, Technology and Science

Spun nanofiber obtained has diameter of 465 nm with 7 % moisture

Average Diameter of Nanofibers a. 139.06 ± 75.02nm b. 217.56 ± 62.74nm

www.irjmets.com @International Research Journal of Modernization in Engineering, Technology and Science

(A) free and (B) encapsulated strain (strain-loaded nanofiber mats)

www.irjmets.com @International Research Journal of Modernization in Engineering, Technology and Science

Entrapment of bacteria increased fibre size up to 400 nm with encapsulation efficacy of 78 %

www.irjmets.com @International Research Journal of Modernization in Engineering, Technology and Science

www.irjmets.com @International Research Journal of Modernization in Engineering, Technology and Science

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