Review Lactic Acid Encapsulation
Review Lactic Acid Encapsulation
Review Lactic Acid Encapsulation
Manel Sebouai1
1
Laboratory of Plant Biotechnology and Ethnobotany, Faculty of Nature and Life Sciences, University
of Bejaia, Algeria
Abstract
Probiotics are live microorganisms that are primarily lactic acid bacteria. They change the
qualities, lactic acid bacteria have long been employed as a functional ingredient in food
vulnerability to harsh and harmful circumstances in the human digestive system, getting into
the human gut alive and in enough cells to exert favorable health effects is still a significant
challenge. Encapsulation has thus been used and researched as a promising and emerging
strategy for good and dependable protection. Therefore, this review focuses primarily on
health potentials of probiotics as well as their bioactive substances. Moreover, the advantages
Introduction............................................................................................................................................2
Health advantages and therapeutic potential of lactobacillus plantarum.................................................4
Probiotics and Immune Response.......................................................................................................5
Inflammatory Bowel Disease (IBD)...................................................................................................5
Anti-cancer activity 6
Effect on Cholesterol 6
Allergies and Probiotics Therapy.......................................................................................................6
Diabetes 7
Active Biomolecules produced by lactic acid bacteria...........................................................................7
Bacteriocin 8
Fatty acid 8
Organic acid 8
Hydrogen peroxide 9
Exopolysaccharide 9
Phenyl lactic acid 10
Enzymes 10
Catalase........................................................................................................................................10
Superoxide dismutase...................................................................................................................10
Glutathione peroxidases 11
Encapsulation of Probiotic Bacteria.....................................................................................................11
Encapsulation techniques used for encapsulating probiotics................................................................13
Extrusion 13
Emulsification 14
Coacervation 14
Spray Drying 14
Freeze drying 15
Electrospinning 15
Matrices used for the encapsulation.....................................................................................................16
Alginate 16
ĸ-Carrageenan 16
Chitosan 17
Gelatin 17
Starch 17
Xanthan 18
Introduction
Functional foods and nutritional interventions usually contain beneficial microorganisms that
can be consumed orally as probiotics. This applies to both people and livestock animals that
are given microorganisms directly to enhance health and reduce pathogen loads.
Food and Agriculture Association of the United Nations (FAO) and World Health
Bifidobacterium species, Saccharomyces boulardii, and other microbes have been proposed
and are used as probiotic strains 2. Due to their long history of use in conjunction with
fermented foods, lactic acid bacteria (LAB) are generally recognized as safe, functional, and
active food ingredients. Their metabolic byproducts, like lactic acid and bacteriocin, can also
be used as natural preservatives and antimicrobial agents to prevent contamination and food
spoilage 3. In general, a probiotic isolated from one animal performs less well in another
animal. However, because many probiotics are of unknown origin, probiotic use across
microorganisms, but the presence of harmful microbes leads to a variety of diseases and
illnesses because toxic substances build up during digestion as a result of the bad microbes.
The immune system begins to weaken at this point. In these circumstances, consuming
probiotics in higher doses is an alternative to using antibiotics to treat the issue 5. According to
studies, probiotics can improve lactose tolerance, lower cholesterol, modulate immunity, and
even prevent some cancers 6. The most used probiotic strains are Lactobacillus and
greater than 107 CFU/mL or g end products at the time of consumption 6. A potential probiotic
strain is predicted to possess several desirable traits to exert its beneficial effects. The
able to function in the gut environment, survive through the upper gastrointestinal (GI) tract,
arrive alive at its site of action and has a reputation for not being pathogenic. Also, it has no
7,8
antibiotic-resistance genes that can be passed on . However, several factors, including pH,
hydrogen peroxide, oxygen, and storage temperature, have been reported to affect probiotic
viability 9. Several approaches for increasing the resistance of these sensitive microorganisms
methods and has received special consideration and investigation. Encapsulation is the
process of retaining cells within an encapsulating membrane to reduce cell loss in a way that
11
results in the release of appropriate microorganisms in the gut . Some advantages of cell
survival during freezing, storage, and converting them into a powder form that is easier to use
The purpose of this article is to review the health advantages and therapeutic potential of
probiotics, the methods and matrices used for their encapsulation and their benefits.
Probiotics’ beneficial effects on human health are diverse and exerted through a variety of
complex mechanisms, and these effects can be local or systemic (extraintestinal effects).
Research suggests that certain variations in the composition of the gut microbiota are linked
13,14
to a variety of diseases . To identify the microbiota imbalance in human diseases like
inflammatory bowel disease or obesity, this was confirmed by comparing the microbiomes of
15,16
healthy individuals with those of diseased individuals . Some of the health advantages and
therapeutic potential of lactobacillus strains are discussed below, with an emphasis on in vivo
The ability of probiotic bacteria to modulate the immune system is well established. The
probiotics can interact with lymphocytes, dendritic cells (DCs), monocytes/macrophages, and
epithelial cells. The majority of pathogens share common structures called pathogen-
associated molecular patterns (PAMPs), which are recognized by the innate immune system.
In contrast, the adaptive immune response, which is highly specific for particular antigens,
17
depends on B and T lymphocytes . Pattern recognition receptors (PPRS) that bind with
(PAMPs). Pattern recognition receptors have been the subject of extensive research.
Typically, these receptors are toll-like receptors (TLRs). Extracellular C-type lectin receptors
(NOD)-like receptors (NLRS) are known to transmit signals upon interaction with bacteria in
18
addition to pattern recognition receptors . The host cells that interact with probiotics are
well-established to be intestinal epithelial cells (IECs). Probiotics may also come into contact
with dendritic cells (DCs), which are important for both innate and adaptive immunity.
Through their pattern recognition receptors, IECS and DCs can both react to gut microbes 17,18.
Probiotics can also alter the quality of intestinal mucins to prevent pathogen binding.
Two idiopathic inflammatory bowel diseases are Crohn's disease and ulcerative colitis. The
gastrointestinal tract as a whole, from the mouth to the anus, can be affected by Crohn's
disease, whereas ulcerative colitis is limited to the colon 19. Patients with IBD frequently use
probiotics, and their doctors frequently suggest them as an adjunctive treatment 20. Although
little is known about the mechanisms underlying probiotics' positive effects, these are
plantarum subsp can be found in the normal gut of healthy individuals as well as in a number
Probiotic lactobacilli may activate anticancer mechanisms in the human gut by either
preventing the carcinogenesis process or directly inhibiting the proliferation of cancer cells, as
24 25,26
suggested by several in vitro studies . while some strains have been demonstrated to
27,28
have powerful immunomodulating properties . Studies have shown that bacterial
byproducts can change the environment in the gut and influence the development of cancer
29,30
. By scavenging reactive intermediates and producing carcinogen-deactivating and
reductase, glutathione peroxidase, superoxide dismutase, and catalase, LAB can reduce the
Effect on Cholesterol
bile acid deconjugation by bile salt hydrolase (BSH), cholesterol assimilation by probiotics,
cholesterol co-precipitation with deconjugated bile, cholesterol binding to probiotic cell walls,
and cholesterol incorporation into probiotic cellular membranes during growth 33.
The possibility of getting a food allergy disease is increasing globally. Preventing allergies
probiotics can help with the treatment and prevention of food allergies 34. Lactic acid bacteria
have been shown to be potential human therapeutic agents. They have been discovered to alter
the human digestive system's intestinal microflora. The term "probiotics" refers to a living
microorganism that promotes health, specifically lactic acid bacteria. Probiotics should be
mother's microbial environment during pregnancy, and how it may influence the offspring's
immune development even before birth. The lack of microflora in babies' intestines during the
early stages of life causes their digestive systems to be disturbed. Perhaps this explains why
pediatricians advise taking a probiotic to restore normalcy to the digestive system. Probiotics
are a good, all-natural alternative for the prevention and treatment of many diseases because
of their immunomodulatory qualities and safety in use. Additionally, allergic patients receive
probiotic therapy for the treatment and future prevention of allergic issues 35,36.
Diabetes
Type 2 diabetes is a major modern disorder that costs governments a lot of money. Individual
health is influenced by genetic and environmental factors. Individuals with this pathology
have milder gut dysbiosis than patients with inflammatory bowel disease. However, there was
a decrease in the proportion of the phylum Firmicutes and the Clostridia class 37. Furthermore,
pathogens. These bacteria have been shown to protect against a variety of diseases 38. Even
more interesting is the increase in intestinal permeability seen in obese and diabetic mice
when fed a high-fat diet 39. Such a diet results in a decrease in the presence of Bifidobacteria,
Different antimicrobial, antiviral and antifungal substances, including organic acids, hydrogen
various Lb. plantarum strains. Due to their potential use in food preservation to increase shelf
life and reduce or even eliminate chemical additives, as requested by consumers, the choice of
and Bifidobacteria. Bacteriocins have a high antibacterial activity and have been used by
humans for thousands of years in the production of fermented foods. Bacteriocins are
classified according to their size, mode of action, and inhibitory spectrum. Bacteriocins have
many beneficial effects, including inhibiting the growth and development of pathogenic
bacteria and being heat and pH resistant. Bacteriocins' antibacterial mechanism is primarily
observed through their effect on bacterial cytoplasmic membranes. Bacteriocins are linked by
affecting sensitive components such as bacterial peptides and inhibiting spore growth and
Fatty acid
Fatty acids are among the most important metabolites of probiotic bacteria. These compounds
have numerous beneficial effects (such as antibacterial and antiviral properties). The antiviral
effect of fatty acids is due to their unique structural properties. Fatty acids are made up of a
saturated and unsaturated carbon chain that is joined by a carboxylic (hydrophilic) group.
Lauric and meristic acids are two fatty acids that are particularly effective against bacterial
growth and development. Short-chain fatty acids (FACs) produced by probiotic bacteria have
become increasingly popular in recent years. Because of their high antibacterial activity, they
Organic acid
Organic acid from probiotics can be used as antimicrobial, antiviral agents. Organic acids are
well-known as important postbiotics. The most important acids produced by probiotic bacteria
are citric acid, acetic acid, and tartaric acid, all of which have strong antibacterial properties.
Lactic acid, acetic acid, tartaric acid, malic acid, and citric acid inhibit pathogens by lowering
pH significantly. Lactic acids' inhibitory effect is related to their effect on bacterial cell
intracellular pH and reducing membrane integrity. Lactic acid can be found in two isomers: L
and D. The L-shape is better at preventing viral infections. Citric acid and acetic acid also
prevent the infection of viruses by creating an acidic environment. The antiviral mechanism
of organic acids produced by probiotic bacteria is observed through the binding of organic
acids to the glycoprotein (S) of viruses, thereby preventing viruses from binding to the
Hydrogen peroxide
Hydrogen peroxide is produced primarily by all bacteria, but it is most visible in aerobic
47
cultures of catalase-negative bacteria and is the major metabolite of lactic acid bacteria .
These substances have potent antiviral properties. These compounds disinfect materials by
interacting with vital components of bacteria, fungi, and viruses, such as enzymes and nucleic
Exopolysaccharide
EPSs are bacterial components produced by lactic acid bacteria (LAB) that have been shown
to play an important role in probiotic activity such as survival, adhesion, and anti-tumor
effect. LAB EPSs have a wide range of sugar compositions, which can be classified as
Enzymes
Catalase
Catalase, a naturally occurring enzyme found in probiotics and other living organisms,
52
degrades hydrogen peroxide (H2O2) into oxygen and water . Catalase is a Reductase
Oxidase enzyme that suppresses ROS. As a result, it acts as an antioxidant and protects the
cell from oxidative stress by inhibiting active oxygen species. Catalase is made up of four
polypeptide chains, each of which contains more than 500 amino acids. It has four porphyrins
(iron) in it that allow it to react with oxygenated water. The pH range for this enzyme's
Superoxide dismutase
Superoxide dismutase can catalyze and facilitate the radical decomposition of superoxide
55
(O2) molecules into ordinary oxygen (O2) or hydrogen peroxide (H2O2) molecules . In
living organisms, there are three superoxide dismutase isoforms: cytosolic SOD1 (copper,
which plays a different role in cell health. SOD1 protects the cytoplasm of cells, SOD2
protects the mitochondria of cells from free radical damage, and SOD3, as an antioxidant
diseases. With its antioxidant activity, this enzyme protects tissues from the effects of
oxidative stress 56,57 Superoxide dismutase is a critical enzyme in living organisms' antioxidant
systems. Probiotics, like all living organisms, have this enzyme, which, as a probiotic
Glutathione peroxidases are an enzyme family with peroxidase activity whose main biological
to alcohol, thereby preventing the formation of free radicals, because cellular lipid compounds
are sensitive to free radicals and produce lipid peroxide as a result of the reaction. Glutathione
peroxidase isoenzymes are encoded by different genes 60. These isoenzymes are distributed
throughout the cell and have distinct substrate properties. Glutathione peroxidase 1 is the most
abundant isoenzyme found in the cytoplasm of almost all mammalian tissues, and hydrogen
61
peroxide is its preferred substrate . Glutathione peroxidase 2 is an extracellular enzyme
found in the intestine. Glutathione peroxidase 3 is also extracellular and found in high
Before delivering materials into a system, encapsulation can be thought of as a technique for
enclosing materials into capsules. There are many encapsulation technologies available
enzymes, polyphenols, enzyme parts, antioxidants, and micronutrients inside wall materials.
particle size (between 3 and 800 µm) (from 10 to 1000 nm). In addition to protecting
those substances directly to the target site, increasing their bioavailability 63.
Extrusion
The most popular method is extrusion because of its simplicity, low cost, and gentle
64
formulation conditions that ensure high cell viability . A hydrocolloid solution is simply
made, microorganisms are added to it, and then the cell suspension is extruded through a
syringe needle in the form of droplets to fall freely into a hardening solution or setting bath.
The diameter of the needle and the length of free fall, respectively, determine the beads' size
Emulsification
emulsifier and a surfactant are required for encapsulation in an emulsion. Emulsifiers are used
to make a more effective emulsion because they reduce surface tension, resulting in smaller
66–69
particles. The emulsion is then treated with a solidifying agent (calcium chloride) . The
emulsion technique has a high rate of bacterial survival and is simple to scale up 67.
Coacervation
varying the pH, ion concentration, matrix molecule-to-bioactive component ratio, and matrix
type. The technique is primarily driven by electrostatic interactions, but it also involves
encapsulation technology, it is mostly proposed and used for bioactive food molecules rather
than bioactive living cells. The method is used for flavors, oils, and some water-soluble
Spray Drying
Spray drying is one of the industrial encapsulation techniques that are most frequently used.
Both living probiotics and bioactive food molecules are being used in their application. When
properly carried out, it is a quick and reasonably inexpensive procedure that is also very
reproducible. The core is dissolved in a matrix material dispersion according to the spray
drying principle. After that, the dispersion is atomized in heated air. This encourages the
solvent to be removed quickly (water). Then, at the outlet, the powdered particles are
separated from the drying air at a lower temperature. The widespread use of spray drying in
industrial settings is primarily due to its relative simplicity and low cost. The technology does,
however, have some significant drawbacks. The first is its limited range of use. Since it is an
might be exposed. This is particularly troublesome when taking into account the method used
to encapsulate probiotics, as the bacteria may leak into the product when some hydration takes
place. Another drawback is the required high temperature for immobilization, which is
incompatible with the survival of all probiotic species. Bifidobacteria, for instance, was found
Freeze-drying is a process that involves freezing the probiotic bacteria, usually in the presence
increase microorganism survival and stabilize the product during storage. The lack of
oxidation and water phase transition is the main advantage of this technique. This greatly
increases the probiotic bacteria's survival rate. In comparison to spray drying, the equipment
Electrospinning
create a jet, which is then stretched and elongated to create fibers 72. It has been used as an
alternative encapsulation method for sensitive foods and bioactive substances in recent years
73
due to its numerous advantages . It is a typical process for producing nanofibres with
diameters of 100 nm or less 74, which was initially introduced by Formhals in 1934 75.
Alginate
acids that are extracted from various algae species. The amount and sequential distribution of
the polymer chain vary depending on the source of the alginate, and this influences the
widely used in the encapsulation of cells because it is simple, non-toxic, biocompatible, and
inexpensive. Another benefit is the solubilization of alginate gels in the human intestines by
trapping calcium ions and releasing trapped cells. The sodium alginate and calcium chloride
ratios used to make the beads range from 1 to 3% alginate with 0.05 to 1.5 M CaCl 2 67,76. The
benefit of alginate is that it can easily form gel matrices around bacterial cells, it is safe for the
body, it is inexpensive, mild process conditions (such as temperature) are required for its
performance, it can be easily prepared and properly dissolve in the intestine and release
entrapped cells. However, there are some drawbacks to using alginate beads. For example,
alginate microcapsules are sensitive to acidic environments, which is incompatible with the
that the microbeads produced are very porous, making it difficult to protect the cells from
ĸ-Carrageenan
ĸ-carrageenan is a natural polymer widely used in the food industry. The cells must be added
using the compound. The gelation takes place when the mixture is cooled to room
temperature, and the microparticles are then stabilized by adding potassium ions.
Chitosan
Chitosan is a glucosamine-based linear polysaccharide that can be extracted from fungi, insect
cuticles, and crustacean membranes. It can polymerize by forming a cross-link when anions
and polyanions are present. This ingredient has not demonstrated a strong ability to increase
cell viability through encapsulation, and it is best used as a coat rather than a capsule.
Probiotic bacteria can be delivered to the colon in a viable state by being enclosed in an
alginate and chitosan coating, which offers protection under simulated GI conditions 78,79.
Gelatin
Gelatin is a protein that is formed from the partial hydrolysis of collagen. It has a unique
structure and versatile functional properties, and it forms a viscous solution in water that cools
to form a gel. Because of its amphoteric nature, it can have synergistic effects with anionic
polysaccharides such as gellan gum. At pH greater than 6, the two polymers mentioned are
miscible because they both have net negative charges and repel one another. When the pH is
reduced below the isoelectric point of gelatin, the net charge on the gelatin becomes positive,
diisocyanate mixture produces strong capsules that are resistant to cracking and breaking,
especially at higher concentrations. This is due to the formation of cross-links between these
polymers 69.
Starch
structurally distinct molecules: amylose and amylopectin. Amylose refers to the linear and
helical chains of glucose polymer, whereas amylopectin refers to the highly branched chains.
The content of each fraction varies depending on the origin of the starch, but in general, it
contains 20-30% amylose and 70-80% amylopectin 77. The starch granule is an ideal surface
for probiotic cell adhesion, and resistant starch which can not be digested by pancreatic
80
enzymes in the small intestine, can reach the colon and be fermented . As a result, the
resistant starch has a good enteric delivery characteristic, resulting in a better release of
bacterial cells in the large intestine. Furthermore, resistant starch can be used by probiotic
bacteria in the large intestine due to its prebiotic functionality. It has been shown to be
Xanthan
pentasaccharide units made up of two glucose units, two mannose units, and one glucuronic
acid unit. The polysaccharide is made by fermenting the bacterium Xanthomonas campestris
and then filtering or centrifuging it. This polymer is soluble in cold water and quickly
hydrates. Even though xanthan is primarily non-gelling, a mixture of xanthan and gellan gum
Conclusion
In general, identifying and selecting probiotics is a safe choice for ingesting drugs and other
therapies to control health imbalances in humans and animals. Probiotics are protected by
conditions, making them one of the most efficient ways to maintain viability and stability.
New materials are being tested, and new technologies, like electrospinning, are being
developed, in addition to the polysaccharides that have traditionally been used as a matrix in
apparatus are needed. More research is also needed to identify appropriate carrier matrices
anticipated in order to be minimized. Simpler methods, less bacterial waste, and greater
product health effects can all result in cost reductions. The research, on the other hand,
intends to promote the usage of encapsulated probiotics in various food matrices.
Furthermore, few in-vivo studies have been conducted to investigate the beneficial effects of
probiotics that have been encapsulated. Despite the excellent results, these research were only
conducted on animals. Large-scale clinical trials involving a large number of patients will be
required to obtain solid proof of the preventative and therapeutic efficacy of encapsulated
probiotics in medical practice. These microorganisms' ability to colonize their niche, as well
as information on the optimal formulations in terms of bacterial viability and number, will be
required. The administration schedule must be standardized in order to get uniform and
comparable results.
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