ISSN: 0973-7510

E-ISSN: 2581-690X

Review Article | Open Access
Priya1, Poonam Joshi1 , Jaya Rautela2, Pallavi Pandey1, Srishti Morris1 and Pallavi Ghildiyal1
1Uttaranchal Institute of Pharmaceutical Sciences, Uttaranchal University, Dehradun-248007, Uttarakhand, India.
2Department of Pharmaceutical Sciences, School of Health Science and Technology, UPES, Dehradun, Uttarakhand, India.
Article Number: 9302 | © The Author(s). 2024
J Pure Appl Microbiol. 2024;18(2):853-866. https://doi.org/10.22207/JPAM.18.2.52
Received: 08 February 2024 | Accepted: 09 May 2024 | Published online: 02 June 2024
Issue online: June 2024
Abstract

Salt Tolerant Microbes are a group of microorganisms that grow, develop, and survive in extremely high salt concentrations. Based on their tolerance level they generally grow up optimally at pH values beyond 9.0, but the growth is inhibited at the pH value that is most closely associated with neutral 6.5. They have minimal dietary needs and a good salt quantity that is high enough to osmotic pressure. They can produce biological metabolites that have certain actions such as antibacterial, antifungal, antioxidant, and anticancer. We discussed in this article various pharmaceutical formulations of salt-tolerant microbes, every formulation shows the specific pharmacological actions like anti-cancer activity, anti-oxidant activity, and anti-microbial activity, and also discusses methods for the biosynthesis of salt-tolerant microbes’ nanoparticles.

Keywords

Salt Tolerant Microbes, Nanoparticles, Metabolites, Microorganism

Introduction

Salt-tolerant microbes are divided into three main categories i.e., Mild, Moderate, and High. Salt Tolerant Microbes require levels of sodium chloride above 3% in salt water.1 Several factors affect the tolerance criteria like growth medium, pH, temperature, and salt concentration. They generally grow up optimally at pH values beyond 9.0, but the growth is inhibited at the pH value that is most closely associated with neutral 6.5.2,3 The general mechanism followed by Salt Tolerant microbes for surviving in varying saline conditions.

They have minimal dietary needs and a good salt quantity that is high enough for osmotic pressure.4 The processes of their halo adaptation (halophilic bacteria can maintain growth and development under salinity conditions) on the internal storage of KCl (potassium chloride) approximately 37% or maintain the equilibrium of sodium ions in the cell plasma and resist the osmotic pressure of the outside surroundings caused by the excessive salinity.

These organisms act on an osmoregulatory mechanism that helps them to grow in acute concentration by balancing ion exchange with the surroundings and their tolerance along with surviving for a long-life span brings the Salt Tolerant Microbes into a new era of development.5-8

Hypersaline regions offer several possibilities for the production of additional metabolites with industrially relevant bioactivities.9,10

The secondary metabolites produced by the Salt Tolerant Microbes show properties against drug-resistant bacteria. Using these metabolites with the help of nanotechnology will be used as a powerful tool in pharmaceutical sciences.11-13

They can produce biological metabolites that have certain actions such as antibacterial, antifungal, antioxidant, and anticancer. The overuse of basically antibiotic drugs has resulted in the development of drug resistance (DR), which reduces or eliminates their efficacy.14,15 The DR is shown in the human body (Cancer cells and micro-organisms). So, to overcome DR issues advanced technologies have been introduced example drug-resistant strains using biomaterials and metabolites. The challenge is to overcome the difficulties related to drug resistance not only in animals and environmental aspects but also in humans.  Two types of genes are responsible for resistance, where the first is horizontal gene transfer (HGT) and the second is genes that are already encoded in the bacterial genome that can give antibiotic resistance by mutation and activation of mobile elements. The recent drug resistance example is third-generation cephalosporin caused by a mutation in genes encoding penicillin-inactivating enzyme. The invention of new medications is necessary to overcome DR issues for better treatment and the Salt Tolerant Microbes (active metabolites) nanoparticles attracted a lot of attention. Some examples of DR have been observed in clinical strains against various antibiotics, affecting both gram-negative and gram-positive bacteria. Resistance is perceived in Haemophilus influenza to ampicillin, in Helicobacter pylori to clarithromycin and in Staphylococcus aureus shows intermediate resistance to vancomycin. Pseudomonas aeruginosa and Neisseria gonorrhoeae to aminoglycosides and quinolone, Enterobacteriaceae to cephalosporins and carbapenems. Furthermore, Enterococcus faecium exhibits resistance to cephalosporin and vancomycin, whereas Streptococcus pneumoniae discerns resistance to penicillin. Unexpectedly, the quorum-sensing mechanism of Pseudomonas aeruginosa led to the development of fluconazole resistance in Candida albicans. In response to these challenges, there has been a growing interest in utilizing halophilic biomolecules to combat DR bacteria.16-20

Nano formulation of existing salt tolerant microbes for the targeted diseases
Several factors are required for the biosynthesis and formulation of bioactive metabolites for example uniform size, high purity, and composition for synthesis through selected techniques.21,22

Various existing nanoformulations of Salt Tolerant Microbes with different targeted delivery like anticancer, antimicrobial, and antioxidant are discussed in Table 1.

Table (1):
Various nano-formulations of Salt Tolerant Microbes

Targeted delivery
Salt Tolerant Microbes
Nano-formulations
Ref.
Anticancer
Archaeal halophile Haloarchaea
Halomonas elongata.
Ideomarina species
nanoparticles synthesis, and gas vesicles
Silver nanoparticles
Selenium nanoparticles (antibacterial and antioxidant)
Selenium nanoparticles
29
Antimicrobial
Halophilic Archaea
Chromohalobacter salexigens, Halobacillus halophilus, and Halomonas elongate
Archaeal Salt Tolerant Microbes
Halophilic archaeon
Extremophilic bacterium-A30
Archaebacteria, actinomycetes, cyanobacteria, and fungi.
Encapsulation of carotenoids isolated from halophilic Archaea in oil-in-water (O/W) nano- and micro-emu
Ectoine nasal spray
Nanoparticles synthesis, and gas vesicles
Gas vesicle nanoparticles
Silver nanoparticles
Silver nanoparticles
30,31
Antioxidants
Streptomyces Marietta, Bacillus subtilis, Bacillus tequilensis, and Bacillus Haynes
Haloferax volcanii BBK2, Haloarcula japonica BS2, and Halogeometricum borinquense E3
Methanolic extract of halophilic bacterial strain
Nanoneedles and selenium nanospheres
32,33,34

Different methods for preparing nanoparticles with key findings are discussed in Table 2.

Table (2):
Methodology and Key Findings (application) of Biosynthesized Nano-particles

No.
Methods
Key- findings
Ref.
1
Dynamic aggregation with radiation-induced cross-linking
Drug carrier
35-39
2
Genetically encoded synthesis in E. coli
Treatment of cancer-conjugated drug
40, 41
3
Electrospraying
Doxorubicin drug delivery model: ph-responsive drug
42-46
4
Ionic gelatin
Delivery of hydrophobic bioactive compounds
47
5
Redox reaction
Anti-cancer treatment
48
6
One pot synthesis
Platinum drugs delivery to cancerous cells
49, 50
7
Lyophilization
Treatment of breast cancer
51
8
Ring-opening polymerization
Drug delivery for prostate carcinoma
52
9
Self-assembly
Biosensors
52

Currently, the nano-formulation of Salt Tolerant Microbes’ active metabolites is considered a scientific tool in the pharmaceutical field. The current approach is focused on treating antibiotic resistance.  The study suggests that the metabolite of Salt Tolerant Microbes produced due to the lack of nutrients, starvation, dehydration, UV-rays, and imbalanced ion concentration makes them deserving candidates for drug discovery. Some examples are also discussed like Anti-cancer activity (Table 3), and anti-oxidant activity (Table 4) in the table we also discuss some tests of anti-oxidant activity like DPPH stands as 2,2-diphenyl-1-picrylhydrazyl is a chemical compound commonly used to measure the antioxidant properties of a substance. The principle behind the DPPH assay is to determine antioxidant properties, through which antioxidant substances will neutralize the free radical of DPPH. ABTS stands for (2,2-casino-bis (3-ethylbenzothiazoline-6-sulfonic acid)), and it measures the ability of antioxidants to scavenge free radicals and prevent oxidation. It observed the reduction of the blue-green color of the solution. The nitric oxide test is used to measure the antioxidant activity of a substance. Nitric oxide is a free radical on which different substances are tested to determine their antioxidant properties by using various methods such as the Griess reagent method.

Table (3):
Anti-cancer activity of a few Active Metabolites of Salt Tolerant Microbes with targeted deliveries

No.
Salt Tolerant Organism
Metabolite
Targeted delivery
Ref.
1
Bacillus species
3-Methyl-2(2-isopropyl) furan
Cervical carcinoma
53
2
Nocardiopsis species HYJ128
Borrelidin C, Borrelidin D
Stomach and leukemia carcinoma
54
3
Streptomyces sp.
Salternamide A
Colorectal and gastric cancer
55
4
Streptomyces species. WH26
Naphthomycin
Lung adenocarcinoma, cervical carcinoma
56
5
Nocardiopsis species
Methyltetrangomycin
Liver cancer
56

Table (4):
Active metabolites of Halophilic micro-organisms responsible for anti-oxidant activity

Salt Tolerant Microbes Structure/ metabolite Antioxidant assay Free radical cleavage capacity Reference
Nocardiopsis gilva YIM 90087 DPPH/ ABTS 54.9 ± 2.0% at t 2 mg/ mL in DPPH 68.6 ± 1.0 at 1 mg/mL in ABTS 57
DPPH / ABTS 14.3 ± 1.5% at 4 mg/mL
28.4 ± 2.7 at 2 mg/mL in ABTS
DPPH / ABTS d 47.7 ± 0.9% at 2 mg/mL
78.2 ± 3.7 at 0.5 mg/mL in ABTS
ABTS 54.6 ± 0.6% at 2 mg/mL
Halophilic Archaea C50 carotenoids EPR spectroscopy 43.17 ± 1.79 of microemulsion and 66.08 ± 0.25 of Nanoemulsion (5 min) 82.38 ± 0.13 of microemulsion and 87.98 ± 2.13 of Nanoemulsion (30 min) 57
Haloterrigena turkmenica Carotenoids (BR, MABR, and BABR) DPPH / FRAP 66.8% ± 1.2 with DPPH 0.067 ± 0.008 at 0.2 µg with FRAR 57
Halomonas nitroreducens strain WB1 EPS (glucose, mannose, galactose, rhamnose, arabinose) Hydroxyl radical scavenging activity, DPPH radical scavenging activity 5.0 mg/ml show 83.3% 58
Phialosimplex species DPPH, OH, b-carotene antioxidant 450.92 ± 6.73 (mg/ml), 390.97 ± 3.97 (mg/ml), 36.59 ± 2.94(mg/ml) respectively 58
Nesterenkonia species HJ01pe DPPH, ABTS 37.7 ± 1.7 and 16.2 ± 0.5 respectively 58
Staphylococcus Arlette
Bacillus subtilis

Bacillus tequilensis
Bacillus
Hayne
ABTS, nitric oxide, phosphomolybdenum, FRAP, and DPPH 86.34 ± 0.007 mM TE/g extract with ABTS 64.79±0.004 μg/ml with nitric oxide 1.69 ± 0.024 mg AAE/g extract with phosphomolybdenum 28.19 ± 0.012 mM Fe (II) E/mg with FRAP 5.48 mg/ml with DPHH 59
Haloarcula species ABTS 95.6 ± 0.1 at 200 004 μg/ml 59
Halorubrum species HRM-150 bacterioruberin (84.12 %), followed by mono anhydrobacterioruberin (15.13 %) and carotenoid extract 2,2-azinobis-3-ethyl-benzothiazole-6-sulfonic acid, 2,2-diphenyl-1-picrylhydrazyl, and hydroxyl radicals. Bacterioruberin shows better antioxidant activity 59
Aspergillus terreus Tsp22
Aspergillus flavus, Aspergillus gracilis, Aspergillus penicilliosis
Crude extracellular compounds  60
Halogeometricum limi strain RO1-6
Haloplanus vescus strain RO5-8
1,1-diphenyl-2-picrylhydrazyl radical scavenging assay. 60

Anti-microbial activity (Table 5) of a few Active Metabolites of Salt Tolerant Microbes with targeted deliveries. The production of carotenoid colors, retinal amino acids, hydrolytic enzymes, and suitable solutes like macromolecule stabilizing agents, biopolymers, and biofertilizers by halophilic microbes is well known. The production of bioplastics, artificial retinas, photoelectric devices, holograms, biological sensors, and other products uses salt-tolerant microbes and extremely halophilic aerobic archaea, also known as haloarchaea, plays a significant role in the industry.23-28

Table (5):
Salt Tolerant Microbes metabolites examples with antimicrobial activities

No
Salt Tolerant Microbes Organisms
Metabolite
Activity
Ref.
1
Nocardiopsis species
Antibacterial
61
2
Bacillus subtilis
Antibacterial, antifungal
61
3
Aspergillus flocculosus
Antibacterial
61
4
Bacillus subtilis
Glycoprotein
Antibacterial, antifungal
62
5
Norcardiopsosis terrae
Antibacterial and anticancer
62
6
Halomonas salaria
Amylase
Protease
8-anilinonaphthalene-1-sulfonic acid  (lipase) (2S,3R,4S,5S,6R)-2-[(2R,3S,4R,5R,6S)-4,5-dihydroxy-2-(hydroxymethyl)-6 [(2R,3S,4R,5R,6R)-4,5,6-trihydroxy-2-(hydroxymethyl)oxan-3-yl]oxyoxan-3-yl]oxy-6-(hydroxymethyl)oxane-3,4,5-triol  (cellulase) Pectinase DNAase
Antimicrobial
62
7
Actinomyces species
Antimicrobial
62
8
Bacillus subtilis
Carotenoids
Polyhydroxy alkanoates
Antibacterial
63
9
Halomonaselongata
Antimicrobial
63
10
Halobacilluskarajiensis
Peptide furanomycin
Antimicrobial
63
11
Vibrio parahaemolyticus B2
Antimicrobial
63
12
Streptomyces species B6921
Antimicrobial
64
13
Actinomadura species M048
Antimicrobial
64
14
Streptomyces nodosus NPS007994
Antimicrobial
64
15
Steptomyces chibaensis species AUBN1/7
Antimicrobial
64
16
Vibriosp. A1SM3-36-8
Antibacterial
65
17
Pseudonocardia endophytica VHK-10
Antimicrobial
66
18
Halobacilluskarajiensis, Alkalibacillus almallahensis
Peptide furanomycin
Antimicrobial
66
19
Steptomonosporaalba
Antibacterial
67
20
Streptomyces species CNQ-418
Marinopyrroles A
Marinopyyroles B
Antibacterial
67
21
Marinispora species NPS12745
Antimicrobial
68,69
22
Streptomyces hygroscopicus BDUS 49
Antimicrobial
69

 

Future perspective
Public health is currently at risk due to the widespread development of antibiotic resistance. Millions of lives have been saved by antibiotics over the years, yet overuse of these drugs has resulted in the development of multi-drug resistant (MDR), which reduces or eliminates their efficacy. Antibiotic resistance has recently reached critical levels, indicating a rise in fatality in the healthy community and a near-term concern for hospitalized patients. In actuality, problems with MDR infections can cause the majority of patient deaths. The exploitation of all organic and environmentally friendly assets, including major environments as an opportunity for new antibiotic discoveries, is being prompted by the urgent need to develop new antimicrobial medicines that are effective and safe. In 1982, Rodriguez-Valeraetal reported the discovery of the first antibacterial substances produced by salt-tolerated microbes. Halocin is the name given to compounds released by numerous Halobacterium species that can lyse and kill the local microbiota. Haloarchaea synthesize peptides and antimicrobial peptides (AMPs) known as halocins. Although some halocins play an ecological and environmental role, less research has been done on how they interact with human infections.70

The clinical significance of salt-tolerated microorganisms is rarely described in the race against time, and antimicrobial intervention against the most significant risk category of human pathogens is lacking. Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Enterococcus faecium, and Pseudomonas aeruginosa are still potential pathogens.71

In the generation of advancement, nanotechnology has been proven as the speedily evolving field that deals with the size of material 1 to 100 nm in diameter.  biomolecules like proteins, nucleic acid, lipids, and polysaccharides are proven to be found in nature and can be used in the development of nanoparticles because they possess their individual properties. To develop distinctive works of art and purpose biomolecules can be engineered with nanoparticles which can result in a Novel Biomolecule Nanoparticle hybrid. So far, there are limited articles available for biomolecule nanoparticles and Salt Tolerant Microbes that can be persuaded for the creation of drug-resistant bacteria to make a variety of antibacterial drugs and anti-cancer drugs. hence, contemporary information on the technology and current trends in an individual field is required. the present review is about the Salt Tolerant Microbes and the metabolites formed by them and integrating them with nanotechnology to form biological nanoparticles which can be considered as new trends in the field of medicine and pharmaceutical field.72

CONCLUSION

As this review proceeds, the study focuses on the Salt Tolerant Microbes, their origin, and the mechanism through which they survive in extreme saline conditions. along with they also produce bioactive agents with significant uses and applications in the pharmaceutical and healthcare area. Salt Tolerant Microbes show diverse significance in the production of bioactive compounds that show therapeutic properties such as antioxidant, antimicrobial, and anti-cancer. Due to their more distinctive properties, these can be considered novel drugs. A further highlight is to focus the beam toward the integration and conjugation of these bioactive substances with nanoparticles for novel perception. The nanotechnology to form salt-tolerant microbes-mediated nanoparticles can be considered a new trend in the pharmaceutical field. This study will also be helpful in the MDR of antibiotics in the pharmaceutical field.

Declarations

ACKNOWLEDGMENTS
None.

CONFLICT OF INTEREST
The authors declare that there is no conflict of interest.

AUTHORS’ CONTRIBUTION
All authors listed have made a substantial, direct and intellectual contribution to the work, and approved it for publication.

FUNDING
None.

DATA AVAILABILITY
All datasets generated or analyzed during this study are included in the manuscript.

ETHICS STATEMENT
Not applicable.

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