A Brief Review on Lysozyme’s Pharmacology and Drug-Carrying Capacity

 

Punit Dilip Varma^1,2, Yogita Deoprasad Shahu^1,3, Subhash Yende¹, Sumit Kishan Arora¹,

Pranali Mishra⁴, Shailesh Jain⁵, Ankit Mishra⁴*

¹Gurunank College of Pharmacy, Kamptee Road, Nagpur, MS 440026.

²Department of Medicinal Chemistry, National Institute of Pharmaceutical Education and Research,

SAS Nagar, Mohali, PB 160062.

³Poona College of Pharmacy, Bharti Vidyapeeth Deemed University, Pune, MS 411038.

⁴Faculty of Pharmacy, VNS Group of Institutions, Neelbud, Bhopal, MP 462042.

 

ABSTRACT:

In recent years, the repurposing of drugs has been one of the exciting work areas for pharmaceutical scientists worldwide. We have prepared a review on Lysozyme, which will help scientists in this area review its properties. Lysozyme is an endogenous enzymatic peptide present in almost every living thing. It has wide therapeutic uses, including antibacterial, antiviral, anti-inflammatory, and immunomodulatory effects. It is showing prominent uses in various diseases, alone or along with other drugs. It is also used as a drug carrier for kidney targeting. It is categorized as generally referred to as safe by USFDA and EC. It is available on the market as oral formulations. Its traditional production by chicken egg is now a day swapped by recombinant production technologies, including transgenic animals. It has a great potential to be studied for various other activities. This review will help the researchers in selecting the medicament for further scientific evaluations.

 

 

1. INTRODUCTION:

Lysozyme (Lyz) is a significant inborn host safeguard protein, which is also called muramidase or N-acetylmuramide glycohydrolase. They are widely distributed and present in almost all developed living things, including single-cell organisms like bacteria. It is an inherent part of the immune system, which provides excellent antimicrobial, antiviral, antitumor, and immunomodulatory actions.  It is an antimicrobial protein that has a host defensive property. The antimicrobial capacity of Lyz is combined with a significant immunomodulatory role since it can cause alterations in inborn immunity. Lyz can be obtained from various sources such as birds, plants, rodents, insects, viruses, and mammals. Rich quantities of Lyz are present in various bodily secretions such as tears, salivation, milk, and bodily fluid. It is likewise present in cytoplasmic granules of the macrophages and the polymorphonuclear neutrophils. Out of all these sources, hen eggs and animal milk are used commercially to obtain it. Due to technological advances, recombination is also used to get the bulk quantity of it. The Lyz obtained from a human is more dynamic than that obtained from the hen egg white.

 

Lyz is also utilized to develop valuable materials by catalysis;  these metabolic products have various biomedical and other applications. Lyz can be administered in the body via various routes such as oral and intranasal as tablets and sprays, respectively. It is also available in powder for injection form for veterinary application. The presented review focuses on compiling recent research done in the Lyz field to help the researchers torch their path of investigation. Figure1 presents a graphical view of this review article.

 

 

Figure 1: A brief review of Lysozyme’s pharmacology and drug-carrying capacity

 

 

2. HISTORY:

a. Discovery:

Sir Alexander Fleming found lyz in the year 1928. It was first seen during some examination made on a patient experiencing intense coryza. The patient’s nasal discharge was cultured each day on plates made up of blood agar, and for the initial three days of infection, there was no bacterial development except for an intermittent staphylococcal state. From that point, Fleming’s remarkable bacteriolytic agent became known.¹ For, past 90 years, this exceptional protein has gotten the interest of researchers due to its various properties. The hen Lyz was the first protein in which all the 20 usual amino acids are present together, and it was the first Lyz submitted for a complete X-beam crystallographic investigation. It is likewise the primary catalyst for which a definite mechanism of action is proposed.²

 

b. Origin and evolution:

Lyz is an old protein that was supposed to be originated 400 to 600 million years ago. Lyz was initially a bacteriolytic defensive agent and has been adjusted to serve stomach-like functions, i.e., degradation and digestion of components. This stomach-like function was observed on a minimum of two categories of Lyz, c-type, and g-type. ‘c’ stands for chicken type while ‘g’ represents goose type. More commonly present Lyz is c-type, g-type Lyz is generally present in sources other than c-type. Both c-type and g-type share a lower level of homology regarding amino acid sequence still have common secondary and tertiary structures.³ After gene duplication 300 to 400 million years back, there was a rise in the Lyz c gene, which now codes for alpha-lactalbumin, a protein only seen in the lactating mamillary gland. The protein alpha-lactalbumin has a similar structure and gene organization. In a sequence of amino acids, it shares just about 40% character with c-Lyz. Though structurally, they are similar, functionally, they are different. Due to timelined substitutions of specific amino acids in Lyz, the enzymatic activity of alpha-lactalbumin reduced, and the alpha-lactalbumin procured the features essential for synthesizing lactose.⁴

 

3. Types and sources:

For the protection of various prokaryotes and eukaryotes from microbial invasion, Lyz is present inside them. Lyzs exist in the following types based on sources from which they are obtained:

 

i. C-type:

Lyz came into the limelight in the 1920s after the discovery by Fleming. Moreover, after its discovery, it was further subdivided into different types based on their sources. The c-Lyz exists widely in numerous living beings, including plants, viruses, bacteria, birds, fishes, insects, mammals, and reptiles.^2,5 Human lysozyme (hLyz) also belongs to the c-Lyz type. This hLyz consists of a polypeptide chain with 130 amino acid and have overall a positive charge.³ They are readily found in various bodily secretions and fluids such as blood, tear, and saliva and gain an essential role in host safeguards.⁶ Lyz in human milk can be passed to newborn infants through breastfeeding, which helps set the baby’s intrinsic immunity barrier.⁷ hLyz shows antifungal and antiviral actions.^9,10,11 Also, they are perceived as safe for use in the food industry without any harmful impacts when administered as 5 mg/g body weight.¹² The activity of Lyz found in this type is equal to Lyz 2500-3500 µg/ml when checked by the method of analysis presented later in this paper.

 

ii G-type:

After the Embden goose, this type of Lyz was named as g –type Lyz. In some birds, g-type is more significant, while in others, it is c-type. Hikima et al. (2001) found the existence of g-type Lyz outside the bird species.¹² Any information found from the various databases uncovers the additional g-type Lyz homologs in other vertebrates, including fish, mammals, and amphibians.¹² Lyz varies in their species, but they also vary inside the body as per the tissue of origin. The protein and cDNA sequencing have demonstrated that the stomach-Lyz (sLyz) varies in amino acid sequence from those found in kidney and granulocyte (regular mLyz). Lyz gene active in macrophages after removing one Proline residue. i.e., Pro103.⁶ The activity of Lyz found in this type is equal to Lyz 1000-1300µg/ml. The other types of Lyzs are Invertebrate type (i- type) found in mollusks and insects, Barley type (b type) found in Plants, v-type found in virus, p-type found in Rodents. Table no 1 shows concentrations of Lyz in various tissues and sources.

 

Table No: 1. The concentration of Lysozyme in Tissues and sources.¹⁴

 

4. Structure of Lysozyme:

As mentioned in the muramidase (or N-acetylmuramic hydrolase), Lyz is a little monomeric protein formed by four disulfide linkages among eight cysteine residues in its polypeptide chain. There is a total of 130 amino acid residues in the vital structure of Lyz (cLyz). The c-Lyz has a molecular weight of 14.3 kDa, with four disulfide linkages between 6 Cys–127Cys, 30Cys–115Cys, 64Cys–80Cys, and 76Cys–94Cys.

 

It also contains 6 trp (tryptophan), 3 tyr (tyrosine), and 3 phe (phenylalanine ) residues.^15,16 The structure of Lyz has six trp deposits, amongst which trp-62 and trp-108 are considered the significant fluorophores.¹⁷ The hen egg-white Lyz (HEWL), classified under c-Lyz, has received much consideration in recent years because of its availability and activity.¹⁷ HEWL, which has a pKa below (∼11), has a positive charge due to the presence of 17 positively charged (6 Lys, 11 Arg) and nine negatively charged deposits (7 Asp, 2 Glu) in its structure. Due to its high enzymatic activity, HEWL is considered one of the best sources for studying various interactions with compounds like drugs and metals.^19,20 Figure 2 A & B presents unfolded and folded structures of Lyz.

 

Figure 2(A) Unfolded Lysozyme     Figure 2 (B) Folded Lysozyme²¹

 

5. Recombinant Human Lysozyme:

The human body secretes the natural hLyz in a limited quantity. Few transgenic animals, including mice, goats, bovines, and pigs, have been utilized to give rise to the bioactive recombinant human lysozyme (rhLyz).¹⁶ The rhLyz in milk not only exhibits their role in lactating animals but is also advantageous to intestinal morphology. Thus, to improve the wellbeing of young animals, the rhLyz is used to adjust the intestinal microbiota composition in babies.²² Likewise, rhLyz obtained from transgenic animal milk has the same bioactivity as natural hLyz, demonstrating that rhLyz has been expressed in animal production may substitute natural hLyz later on.²³ Even so, till now, the declaration of efficient rhLyz in poultry is still infrequently accessible. Thus, causing rhLyz in egg white to effectively decrease the cost and present a promising application prospect.²⁴

 

In the present case, the transgenic chickens were produced in which rhLyz were expressed profoundly in the egg white. Thus, the rhLyz showed the same physicochemical properties as natural hLyz, which uncovered chicken eggs’ ability to produce hLyz. Egg white obtained from transgenic chickens showed a stronger antibacterial action in an agar diffusion test. The reports presented by Ma et al. (2010) showed that, in dwarf chickens, the marker for disease resistance is monocyte-macrophage phagocytosis.²⁵This phagocytosis that occurred in transgenic chickens was considered somewhat more vital than the phagocytosis that occurred in non-transgenic ones, which presumably demonstrated that the hLyz is capable of preventing chickens from getting an infection. hLyz is typically found in tears, salivation, and chicken egg white without undergoing any unfavorable susceptible reaction while administering it in humans and animals. Though there is an issue with rhLyz, it shows its action after refolding it because it is expressed in inclusion form. According to the results obtained after ELISA examination, utilization of the lentiviral vector framework was fruitful in the chickens that express rhLyz. Past exploration uncovered that the overexpression of rhLyz in rabbits causes an issue of lactation. In any case, no issues were found for the production of transgenic chickens.²⁶

 

There has been an attempt to mass-produce egg white Lyz through the recombinant DNA method. The egg white Lyz, which was produced from the recombinant strain, has a surprisingly low production rate and is altogether lower than the current egg white Lyz.²⁷

 

6. Pharmacology:

Lyz hydrolyses the β-1,4 glycosidic bond between N-acetylglucosamine and N-acetylmuramic acid in the bacterial cell wall and kills the gram-positive microorganisms. Lyz also has antimicrobial impacts other than its enzymatic activity. The immunomodulatory capacity of Lyz has been appreciated recently.²⁸ Lyz has a neuroprotective action in Alzheimer's disease as it prevents amyloid-beta aggregation.²⁹ This Alzheimer's disease also has neurological indications in COVID-19 severe patients. From mouse and porcine models, it is clear that the Lyz possesses extensive anti-inflammatory action systemically, bringing about diminished immune-driven pathology.³⁰

 

It showed its activity after oral administration in various clinical trials, although data on clinical trials of Lyz are limited. Generally, oral administration of HEWL shows antiviral action against herpes (1g/day), measles, and hepatitis (60–170mg/day for 4–24 weeks) essentially diminished post-transfusion hepatitis occurrence to 8% as compared with 20% in the negative control. It also showed effective therapy of gum infections (750 mg/day) and skin ulcers, enhancing immune responses in cancer patients with suppressed immunity.¹⁶ For the fast resolution of anti-inflammatory applications and maintenance of Lyz levels in serum and stool of premature newborn children with illnesses, 50 mg/l supplementations in milk for 2–3 weeks are given. A combination of 0.2g Lyz and 1.5g bovine lactoferrin diminished enteric dysfunction in Malawian kids every day. No native or systemic negative impacts have been accounted for these human trials.³¹

 

In Eastern Europe, in lung infections, the HEWL has been combined with antibiotics to treat pneumonia and bronchitis in humans without respiratory or systemic toxicity. It is investigated through animal models that Lyz has been administered via aerosols to treat pneumonia. In P. aeruginosa induced pneumonia in hamsters, administration of 1% solution aerosolized hLyz caused a significant reduction in pneumonia, proven histopathologically, alveolar septal apoptosis, neutrophils, and different leukocytes in the bronchiolar lavage fluid just as the expanded movement of Lyz in that fluid.³⁰ On the other hand, it was noticed that the capacity of hyaluronic acid, the main component of the extracellular matrix, is hindered by Lyz that prevents the elastase fiber from injury. By binding to the elastic fiber and hence, in an animal model on inhalation of Lyz, in case of emphysema, airspaces further expanded, which signifies to remain cautious while administering Lyz through the inhalation route in case of similar infection or disease states.^31,32 Table no 2 summarizes the pharmacological uses of Lyz.

 

Table No. 2: Pharmacology of lysozyme³³

 

7. Therapeutic applications of Lysozyme:

Lyz can be used in various fields of foods and pharmaceutical industries. The Lyz obtained from a human source shows its activity on the bacterial membrane.³⁴ It shows antibacterial, antiviral, antifungal activities, immunomodulatory action, and it is also used as a disease marker for different states of diseases. It is recently found that it can show its activity on novel coronavirus, which is currently the demand of the whole world to find a safe, effective and permanent solution for this crisis.³⁵ The therapeutic applications of Lyz are very well depicted in Fig.2.

 

 

Figure 3. Therapeutic uses of lysozyme

 

I.      Lysozyme as an Antiviral:

Lyz’s sufficient measurements have been demonstrated to be viable in treating viral diseases, for example, herpes simplex, herpes zoster, moles, condylomata acuminata, apoptosis, and vaccinia. It was found that human Lyz is an exceptionally dynamic enemy of HIV connected with the central division of human chorionic gonadotropin from pregnant ladies' urine.²⁷

 

•       Anti-HIV:

The anti-HIV action of Lyz is associated with the central part of chorionic gonadotropins in humans. To better understand the molecular mechanism for anti-HIV action, fragmentation and mapping of Lyz were carried out by various scientists. Moreover, it was independent of its muramidase action (further explained in antibacterial activity). hLyz9 and hLyz18 are the two main peptide fragments considered to be dynamic as that of Lyz. The smallest fragment of hLyz- hLyz9 was found to have full anti-HIV action.²⁷ The amino acid substitution experiment revealed the significance of arginine and tryptophan deposits or residues to anti-HIV action. For the anti-HIV action, both tryptophan and arginine, positively charged amino acids with hydrophobic nature, must be present at the c-terminal portion of hLyz18. The peptide fragment hLyz9, which has a remarkable sequence, is found only in Lyzs obtained from mammalian sources.

 

In complete mapping of hLyz, hLyz9 was found to the region of protein different from catalytic site of muramidase. The hLyz9 exists in R-helix form. Subsequently, for intact hLyzs, the activities such as muramidase and antiviral have unique features. It proves that hLyz9 has anti-HIV action but does not have muramidase action. The peptide hLyz9 directly affects viral entry and its replication. The peptide hLyz9 may disrupt viral molecules or inhibit their binding and entry into target cells. The sensitive profiling of host reaction during viral infection and treatment was done by the cDNA microarray technique, which shows the effect of hLyz9 action on HIV-1 infected targeted cells.³⁴

 

•       Anti-herpes:

Reports by Cisani et al. 1984 showed that altered lysozymes have a solid antiherpes-cytopathic (CPE) action in correlating the antiherpes-CPE movement of local and altered lysozymes with that of other antiviral compounds in vitro. The enzymatic action of Lyz does not appear to be vital for this impact, which seems, by all accounts, to be connected to the simple nature of the Lyz molecule. The unmodified HEWL has shown lower antiherpes-CPE activity than the hi-HEWL (heat inactivated-Hen Egg White Lysozyme).³⁶

 

The anti-herpetic activity of Lyz has been reported recently. It gave the idea that Lyz synergistically affects glycyrrhizic acid, which improves the anti-herpetic activity of this molecule. The Lyz can prevent the cell fusion actuated by HSV blocking and, in this manner, the spread of the viral disease. The anti-herpetic properties of Lyz came about freely from its enzymatic action since the heat-denatured Lyz indicated a higher anti-herpetic activity than the native particle.³⁷ Hasselberger has portrayed the antiviral properties of Lyz against flu and viral hepatitis.

 

II.   Antibacterial Activity:

Some gram +ve and–ve microorganisms were susceptible to various purified Lyz obtained from bovine and human milk. The antibacterial activity of the Lyz gives an impression of being intervened through direct bacteriolytic activity, just as using stimulatory action on the phagocytic function of macrophage and PMN leucocytes.³⁴ Various mechanisms are liable for the bacteriostatic and bactericidal actions of Lyz. Lyz acts in association with the complement system and lactoferrin, which prevents certain microorganisms by chelating iron.  Ogundele (1998) reported that the hLyz has an anti-inflammatory effect.^37,38 Modulation of serum complement actuation is done by inhibiting PMN reaction concerning complement-derived chemotaxis.³⁹ Leo et al. (1978) announced Lyz reduces numerous reactions of neutrophils to inflammatory stimulants.^3,40,41

 

•       Bacteriocidal mechanism of lysozyme:

Usually, it is proposed that Lyz can cause hydrolysis of the β-1, 4-glycosidic linkage of peptidoglycans because of muramidase action, the murein layer breaks results in a reduction in rigidity of the cell wall, which then results in bacterial death.⁴² This enzymatic action is responsible for breaking disulfide linkages, allowing Lyz to work as a nonspecific inborn resistance molecule to attack bacterial microorganisms. According to the structure of Lyz, the β-1,4 glycosidic bond of NAM and NAG is in the nearness to the two capable catalytic deposits, Glu35 and Asp52 of Lyz, is responsible for the destruction of the glycosidic bond, even though after the comprehension of the small facts of Lyz enzyme, mechanism of action is not completely known in these many years.⁴³ Though the significant bactericidal action is restricted to the gm +ev organisms, the lipopolysaccharide (LPS) layer is the principal part on the external surface of gm -ve microbes, which works as a defense part, where the external LPS layer obstructs the entrance of Lyz to the peptidoglycan layer of the cell wall.⁴⁴ The Lyz was utilized in combination with potent chemicals (e.g., lactic acid, EDTA), lysis of some of the pathogens and waste microbes can be advanced and persistent, as these chemicals can permit limited expulsion of some of the components of the cell wall of peripheral surface (outer membrane). Accordingly, the entrance of Lyz into the peptidoglycan was enhanced.⁴⁵

 

III. As a disease marker:

The homeostatic articulation of RES (Reticular-endothelial system) was reflected by Lyz activity, which is possibly the essential defense mechanism of cells in contradiction of infection. Experimental discoveries have created proof for Lyz being an index of macrophage efficient status. In recent times, according to the depiction, the serum Lyz is a potential marker of monocyte/macrophage action in rheumatoid arthritis.⁴⁶ The milk Lyz action has been recommended as a potential diagnostic marker for sub-clinical. The bovine milk ordinarily contains a low level of Lyz, 10-15 µg/ml. Higher concentrations of Lyzs, 1-25µg/ml, were found in mastitis milk. Serra et al. (2002) proposed the function of Lyz as the novel prognostic factor in patients who have breast cancer (in humans).^47,48

 

IV. Immuno-modulation:

The immune-stimulant activity of Lyz was suggested and proved by Rymuszka et al. (2005).⁴⁹ They recommended basic and incomplete processed or fragmented parts from peptidoglycans and compounds with a relatively higher molecular weight.⁵⁰ Other literature also referred to the set of commonly prevalent substances, including Lyz, that can apply an adjuvant or immune-stimulating effect by stimulating the growth of antibody counter to a group of antigens, stimulation of delayed hypersensitivity reaction for the same or different type of antigens, augmentation of disease resistance against the viral or bacterial disease and mitogenic effect.⁵¹

 

V.    Use in coronavirus:

The rhLyz medication can prevent and restrain coronavirus, and the power of preventing covid is superior to the power of inhibition as per the test result. The recent invention has discovered new clinical applications to rhLyz aerosol inhalant, with no toxic side effects and a good outlook in medicine.^30.

 

•       Applications of lysozyme

The Lyz was utilized experimentally in pigs during the 2000s. Lyz (similar to human) obtained from goat milk (transgenic) appeared to alter the metabolic process, intestinal uniformity, and GI microflora. Although, improvements in growth because of the Lyz were not noticed. Besides anti-biotics, Lyzs were also utilized to mitigate and cure various bacterial infections, for example, pharyngitis, tonsillitis, diarrhea, and wound infections. And they are also employed for various dermal applications.⁵²

 

It is also used as a transporter for the bacterial cell's specific drug delivery (antibiotics). Lyzs have also been utilized as the content of mouthwashes because they can kill various oral microbes and respiratory infections (when Lyz is incorporated via the aerosolized system).⁵³

 

Lyzs contains RNase A and urinary RNase U, which degrades viral RNA activity against HIV selectively;  later on, they could be utilized for HIV diseases.⁵⁴

 

Lysozyme as a carrier:

Lyz is suitable as a drug carrier in renal drug targeting as it is a low molecular weight protein (LMWP). After glomerular filtration, LMPW does not undergo tubular reabsorption and gets excreted in the urine. Likewise, the conjugates of Lyz can also be used as carriers for targeting the drug to the urinary tract. Lyz can also be reported to be used for the recognition of food samples.^7,9,55 Table 3 summarizes the works done for the drug-carrying capacity of Lyz.

 

Table No. 3: Drugs for which Lyz is reported as a carrier

 

8. Manufacturing of Lysozyme:

Many attempts have been made for the manufacturing of Lyz. Several transgenic animals have also been utilized for the production of bioactive recombinant Lyz. The hen egg is a unique source for large-scale manufacturing of Lyz. the recent manufacturing method for producing microorganism-derived Lyz is widely accepted. This strategy can bring down the production cost of Lyz and can be applied to different industries. The method suitable for mass production of Lyz is using the culture of Thermomonas koreensis. A Gm-ve, non-spore-forming, rod-shaped, motile bacterium, strain Ko06(T), was isolated from soil from a ginseng field in South Korea and was characterized to determine its taxonomic position.⁶¹ The method for producing Lyz from the culture of Thermomonas koreensis (KCTC 12540) can be shortly described as follows -

 

As per current practices, the culture is prepared by KCTC 12540 at pH 6-8 for 10-20 hours at the temperature range of 10-55°C. Inoculation of 1 to 5%w/w was followed by subculture. A single colony-type bacterium gets activated in primary culture, and more cells get cultivated in secondary culture and, Lyz can be obtained fast from secondary culture. The method for culturing secondary culture involves inoculation of culture medium obtained in the primary culture at a concentration of 1-5%, and the time required for secondary culturing is 20-120 hours. The medium utilized can be mesotrophic or oligotrophic medium. Ideally, the R2A medium (Reasoner’s 2A agar medium) can be used, yet it is not restricted. Centrifugation is done for obtaining Lyz.

 

9. Evaluation:

The Lyz prepared by the above method is evaluated by measuring the protein content and action of the Lyz. The evaluation of Lyz is essential after its manufacturing to ensure its quality;  the following means of evaluation are generally used: -

(1) Measurement of protein content

The protein content is measured spectroscopically. To the 100 μl of the supernatant test sample, 1 ml of a Coomassie Brilliant Blue G-250 solution was added, and the mixture was permitted to remain at room temperature for 20-30 minutes, and the absorbance was estimated at a wavelength of 595nm.⁶²

(2) Measurement of lysozyme action

The Lyz activity was quantified by Micrococcus lysodeikticus dried cells (Gm+ve microbes). The substrate solution is prepared by suspending 0.2mg/ml of dry cells of Micrococcus lysodeikticus in 0.1 M sodium phosphate (pH 6.0). 0.2ml of Lyz supernatant test sample is mixed with 1.8ml of the substrate solution, and the absorbance at 450nm is determined at room temperature for 10 minutes at intervals of 2 seconds. The activity of Lyz was determined by dividing the decrease in the linearly decreasing segment of the absorbance reduction curve by time. One unit for every Lyz unit is the sum that causes the decrease in absorbance (0.001 every moment) by deteriorating Micrococcus lysodeikticus dissolved.

The specific activity was determined by dividing the Lyz activity (unit) by protein (mg).⁶³

 

(3) Lyz purification by milk, which incorporates affinity chromatography (heparin-Sepharose), trailed by gel filtration on Sepharose 4B or Sephadex-G50. For purification, also chromatography on hydroxyapatite has additionally been utilized. Lyz action is regularly tested by the lysis of Micrococcus lysodeikticus yet can likewise be estimated by RP HPLC.⁶⁴

 

10. Regulatory status:

Lyz is persistent in both the animal as well as plant kingdom. Presently, egg-white Lyz is an industrially accepted type of Lyz. The main problems that are associated with that of egg-white Lyz are:

·       High cost of recovery

·       Low action and

·       Most significantly, the immunological issues to specific individuals.

 

Its significant expense obstructs the large-scale manufacturing and wide utilization of Lyz, which are nowadays overcome by easy techniques or low-cost methods. Subsequently, hLyz production has picked up significance and transgenic production of Lyz from animals, microorganisms, or plants.

 

The Food and Drug Administration (FDA) is issuing a tentative final rule to amend its regulations to affirm that egg white Lyz enzyme preparation, when labeled by the common or usual name “egg white Lyz” to identify its source, is generally recognized as safe (GRAS)⁶⁵

The following governing bodies approve Lyz: -

 

Table No. 4: Regulatory Status

 

Table No. 5: Marketed trends of lysozyme

 

 

 

Countries like France, Germany, Japan, and the UK have also recognized its safety and approved it.

 

11. Recent trends:

Lyz has a wide range of therapeutic applications, and therefore it is available in different dosage forms, such as a tablet, capsule, nasal spray, and injection for use in different conditions. There are various marketed formulations available in the market;  some are listed in table 5.

 

12. Conclusion and future perspective:

There are several things, which are yet to disclose about this enzyme Lyz. Lyz has excellent antibacterial and antiviral effects. Along with this, it has a good immunomodulatory effect. Lyzs are obtained from various plants and animals, but the most commercial source was C-type Lyz obtained from chicken. Human lysozyme (hLyz) also belongs to this type of Lyz. Human secretes the human Lyz in limited quantity, and therefore the recombinant human Lyz was prepared by recombinant technology through transgenic animals, which is more efficient than the hen egg Lyz. Lyz can also be used for drug delivery to the kidney. This molecule has great potential to be used alone or with other drugs in various therapeutic conditions.

 

13. Future perspective:

1)    As we know, the Lyz has a wide application in various diseases. Hence, there is broad scope for Lyzs containing products in the future.

2)    The formulations of Lyz, which are available in the market, are expensive because manufacturing Lyz is also expensive. Hence, there is a need to develop a method that is of low cost. i.e., cost-effective products, so the products should be available to a large group of people too.

3)    For products of low cost, the Lyz should be prepared through recombinant technology.

 

14. ACKNOWLEDGMENTS:

We acknowledge Dr. S.J. Daharwal, Pt Ravishankar University, Raipur, for his valuable suggestions in the manuscript.

 

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Accepted on 27.01.2022           © RJPT All right reserved

Research J. Pharm. and Tech 2022; 15(12):5886-5894.