Purification and characterization of anti-tubercular and anticancer protein from Staphylococcus hominis strain MANF2: In silico structural and functional insight of peptide.

The present context was investigated to purify and characterize anti-tubercular as well as anticancer protein from fermented food associated Staphylococcus hominis strain MANF2. Initially, the anti-tubercular potency of strain MANF2 was assessed against Mycobacterium tuberculosis H37Rv using luciferase reporter phase assay which revealed pronounced relative light unit (RLU) reduction of 92.5 ± 1.2%. The anticancer property of strain MANF2 was demonstrated against lung cancer (A549) and colon cancer (HT-29) cell lines using MTT assay which showed reduced viabilities. Anti-tubercular activities of the purified protein were observed to be increased significantly (P < 0.05) ranging from 34.6 ± 0.3 to 71.4 ± 0.4% of RLU reduction. Likewise, the purified protein showed significantly (P < 0.05) reduced viabilities of A549 and HT-29 cancer cells with IC50 values of 46.6 and 48.9 µg/mL, respectively. The nominal mass of the purified protein was found to be 7712.3 Da as obtained from MALDI-TOF MS/MS spectrum. The protein showed the sequence homology with 1-336 amino acids of Glyceraldehyde-3-phosphate dehydrogenase from Staphylococcus sp., thus, categorizing as a new class of Glyceraldehyde-3-phosphate dehydrogenase-like protein. The amino acid sequence of the most abundant peptide (m/z = 1922.12) in the purified protein was obtained as 'KAIGLVIPEIDGKLDGGAQRV' and it was identified as peptide NMANF2. In silico tools predicted significant stereo-chemical, physiochemical, and functional characteristics of peptide NMANF2. In a nutshell, protein purified from strain MANF2 can certainly be used as an ideal therapeutic agent against tuberculosis and cancer (lung and colon).

Isolation and screening of Streptomyces sp. Al-Dhabi-49 from the environment of Saudi Arabia with concomitant production of lipase and protease in submerged fermentation.

In this study, Streptomyces sp. Al-Dhabi-49 was isolated from the soil sample of Saudi Arabian environment for the simultaneous production of lipase and protease in submerged fermentation. The process parameters were optimized to enhance enzymes production. The production of protease and lipase was found to be maximum after 5 days of incubation (139.2 ± 2.1 U/ml, 253 ± 4.4 U/ml). Proteolytic enzyme increases with the increase in pH up to 9.0 (147.2 ± 3.6 U/ml) and enzyme production depleted significantly at higher pH values. In the case of lipase, production was maximum in the culture medium containing pH 8.0 (166 ± 1.3 U/ml). The maximum production of protease was observed at 40 °C (174 ± 12.1 U/ml) by Streptomyces sp. Lipase activity was found to be optimum at the range of temperatures (30-50 °C) and maximum production was achieved at 35 °C (168 ± 7.8 U/ml). Among the evaluated carbon sources, maltose significantly influenced on protease production (218 ± 12.8 U/ml). Lipase production was maximum when Streptomyces sp. was cultured in the presence of glucose (162 ± 10.8U/ml). Among various concentrations of peptone, 1.0% (w/v) significantly enhanced protease production. The lipase production was very high in the culture medium containing malt extract as nitrogen source (86 ± 10.2 U/ml). Protease production was maximum in the presence of Ca2+ as ionic source (212 ± 3.8 U/ml) and lipase production was enhanced by the addition of Mg2+ with the fermentation medium (163.7 ± 6.2 U/ml).

Chemical constituents of Streptomyces sp. strain Al-Dhabi-97 isolated from the marine region of Saudi Arabia with antibacterial and anticancer properties.

BACKGROUND: Unlike the terrestrial region, the microorganisms especially actinomycetes groups existing in the marine environment are important sources for the medically important drugs and other active compounds. Considering the importance of natural compounds from the marine actinomycetes, the present study proceeded to identify and characterize promising antibacterial and anticancer actinomycetes from the marine region of Saudi Arabia and to profile the individual chemical components. METHODS: Antimicrobial, anticancer and chemical profiling were performed by broth microdilution, mitochondrial membrane potential assays and GC-MS analysis. Investigations were directed towards the isolation and characterization of active Streptomyces sp. strain Al-Dhabi-97. RESULTS: The obtained results of the morphological, biochemical, physiological and molecular level studies of the isolate Al-Dhabi-97 showed similarity towards the species of Streptomyces. Gram positive bacteria such as Bacillus subtilis, Enterococcus faecalis, Staphylococcus epidermidis and Staphylococcus aureus showed MIC values of 500, 250, 125 and 62.5µg/ml and Gram negative bacteria such as Pseudomonas aeruginosa, Klebsiella pneumonia, Escherichia coli and Salmonella paratyphi reported MIC values of 500, 500, 250 and >250µg/ml in the antimicrobial studies. The results of anticancer studies showed that at 100µg/ml, the extract showed maximum cell growth inhibition and exhibited 2.5% necrosis, 62.2% late apoptosis and 20.8% early apoptosis in COLO 320 DM and VERO cell lines respectively. Chemical profiling of the extract authenticated the presence of constituents such as 1-phenanthrenemethanol (46.64%), phthalic acid, di(2-propylpentyl) ester (26.97%), benzenebutanoic acid (3.37%), podocarp-7-en-3-one (2.68%), and indole-3-carboxaldehyde (1.11%) respectively. CONCLUSION: The present study concluded that Saudi Arabian marine region was a promising area for the identification of medically important natural products producing actinomycetes for antibacterial and anticancer drugs.

Biosynthesis of silver and gold nanoparticles using Musa acuminata colla flower and its pharmaceutical activity against bacteria and anticancer efficacy.

Synthesis of nanoparticles using plant sources as reducing agent has become important, as physical and chemical methods are costlier and affects environment. Hence it is important to develop environment friendly nanoparticle synthesis by avoiding the use of toxic chemicals. The present study aimed to synthesize silver nanoparticles (Ag Nps) and gold nanoparticles (AuNps) using Musa acuminata colla flower and its pharmaceutical activity against extended spectrum beta-lactamase (ESBL) gene producing bacteria and anticancer efficacy. The synthesized Ag and Au NPs were analysed by means of UV-Vis, FTIR, XRD,SEM and EDAX evidenced the bioreduction of Ag+ ions to Ag0 and Au3+ ions to Au0 respectively. Both nanoparticles and flower extracts were studied for antibacterial activity of ESBL gene producing bacteria by disc diffusion and microdilution (Resazurin) method. In vitro anticancer efficacy (MCF-7) and toxicity (VERO) of AgNPs, AuNPs, aqueous extract and ethanol extract of flowers were performed by MTT assay. IC50 value for DPPH analysis was at 390 µg and 460 µg for ethanol and aqueous extract respectively. Total antioxidant content was found be 740 µg/mg and 460 µg/mg for ethanol and aqueous extract. GCMS analysis authenticated the existence of the compounds namely, 9,12-octadecadienoic acid(z,z)- and n-hexadecanoic acid in the crude extract of the samples. Among the samples, AgNPs had best antibacterial activity. AgNPs and AuNPs were confirmed by colour change to reddish brown and ruby red. Further ƛmax were obtained at 474 and 540 nm by UV - visible spectrum. SEM analysis revealed the particle size ranges from 12.6 to 15.7 nm for silver and 10.1 to 15.6 nm for gold nanoparticles. The EDAX spectrum shows a strong signal for elemental Ag and Au at ~ 3 keV and 1.5 keV. The XRD patterns for silver and gold nanoparticles at 36.701, 42.900, 63.281 and 76.398 corresponding to the lattice planes 2.4467, 2.1064, 1.46839, 1.24564 nm and 27.32, 36.7228, 39.56, 42.888, 63.253, 63.253, 65.02 and 76.383 corresponding to the lattice planes 3.262, 2.44530, 2.276, 2.1070, 1.46897, 1.4332 and 1.24585 nm. The IC50 values for MCF-7 and VERO cells were 30.0 µg/ml and 55.0 µg/ml respectively.

Environmental friendly synthesis of silver nanomaterials from the promising Streptomyces parvus strain Al-Dhabi-91 recovered from the Saudi Arabian marine regions for antimicrobial and antioxidant properties.

The therapeutic molecules recovered from the marine biological origin are widely used for the treatment of diverse levels of infections caused by microbial pathogens. In addition, the eco-friendly preparations of nanomaterials together with the secondary metabolites' producing active microbial strains effectively suppress the spreading of the pathogenic bacteria. Considering their importance, the present study evaluated the environmental friendly synthesis of Silver nitrate nanomaterials (SNM) from the active marine Streptomyces strain Al-Dhabi-91 isolated from the Dammam region of Saudi Arabia. The obtained SNM was chemically characterized by various spectroscopic techniques such as UV, XRD, FTIR, SEM, TEM and EDAX; and its biological applications such as antimicrobial properties and antioxidant potential were recorded by DPPH methods. Biochemical and micromorphological studies together with the molecular techniques confirmed that the isolate Al-Dhabi-91 belonged to Streptomyces species. The characterization techniques confirmed that the UV spectrum showed maximum absorption peak at 305 nm indicating the plasmodium characteristics. SEM and TEM analyses evidenced 5-2 nm which are agglomerated, cool to form porous asymmetrical networks. Additionally, the FTIR spectrum showed maximum peak at 1194 cm-1 and 1394 cm-1, confirming the presence of aromatic CH bending and aromatic CC bending in the SNM. SNM exhibited prolific antibacterial activity against Gram negative pathogens, K. pneumoniae (28.33 mm) and E. coli (21.66 mm) respectively. The MIC values of SNM were significant with respect to E. faecalis (125 µg/ml), S. aureus (250 µg/ml), P. aeruginosa (125 µg/ml), K. pneumoniae (500 µg/ml) and E. coli (250 µg/ml) respectively. In addition, the antioxidant potential of the SNM was another value added importance. Especially 50 µg/ml of the nanoparticles showed 33% antioxidant potential; similarly in nitric oxide radical inhibition assay the concentration of 50 µg/ml nanoparticles showed 33% of inhibition potential. Overall, the eco-friendly synthesis of SNM from the marine Streptomyces strain Al-Dhabi-91 was an ideal active source for the treatment of infectious disease and health associated disorders.

Rapid biosynthesis and characterization of silver nanoparticles from the leaf extract of Tropaeolum majus L. and its enhanced in-vitro antibacterial, antifungal, antioxidant and anticancer properties.

Eco-friendly biosynthesis of nanoparticles from medicinal plants as reducing agent has gained importance due to its potential therapeutic uses. In the present study Silver nanoparticles (AgNPs) were eco-friendly synthesized using the leaf extracts of the medicinal plant Tropaeolum majus. The obtained AgNPs were characterized by UV - visible spectrum, FTIR, SEM and XRD which clearly showed the reduction of Ag+ ions to Ag0. In addition, the aqueous and ethanolic extracts were analyzed for phytochemicals and its antioxidant activities. GC-MS spectrum showed the presence of 25 compounds with benzeneacetic acid as the dominant contents. The synthesized AgNPs revealed maximum absorption spectrum at 463 nm and FTIR vibrational peaks at 3357.46, 21,966.52, 2118.42, 1637.27, 658.571 and 411.728 cm-1 respectively. SEM and XRD studies evidenced the nature of nanocrystalline with face centered cubic (fcc) crystal structure. Both AgNPs and plant extracts showed more inhibition activity against Pseudomonas aeroginosa compared to other bacteria with MIC value of 6.25 µg/ml. Antifungal activities was higher for Penicilium notatum with MIC value 31.2 µg/ml. The IC50 values for MCF7 for aqueous extract were found to be 4.68 µg/ml, ethanol extract 7.5 µg/ml, AgNPs 2.49 µg/ml, and doxorubicin 1.4 µg/ml. The IC50 values for VERO cell line for aqueous extract was 8.1 µg/ml, ethanol extract with 6.8 µg/ml, silver nanoparticles 5.3 µg/ml and doxorubicin 2.6 µg/ml respectively. Conclusively, the antibacterial, antifungal, antioxidant and anticancer properties of the synthesized AgNPs from Tropaeolum majus act as major therapeutic drug for microbial infectious disease and other health associated disorders.

Green biosynthesis of silver nanoparticles produced from marine Streptomyces sp. Al-Dhabi-89 and their potential applications against wound infection and drug resistant clinical pathogens.

Green biosynthesis of nanoparticles yielded from the extracellular metabolites of marine derived actinomycetes is covering enormous attention due to their superior applications and broad spectrum of bioactive secondary metabolites. The present study aimed to explore the characterization of novel bioactive Streptomyces sp. Al-Dhabi-89 derived from the Arabian Gulf region of Dammam was authenticated based on their physiological, biochemical characterization and molecular level amplification of the 16S rRNA was guided for the green biosynthesis of silver nanoparticles from the cell free extract. The reducing potential of Streptomyces sp. Al-Dhabi-89 cell surface extracts was explored for the green synthesis of the silver nanoparticles without the supplementation of the external capping substance, were characterized by UV-Vis, XRD, SEM, TEM, EDX, FTIR, and antimicrobial activities were determined by performing broth micro-dilution techniques. Results indicated that UV-Vis absorption spectrum of the bio-reduced nanoparticles documented the major peak at 310 nm confirmed the silver nanoparticles surface plasmon resonance. TEM and SEM characterization authenticated the morphological size ranged from 11 to 21 nm and cubic in their appearances; in addition, FTIR spectrum revealed variation in the band values from 626 cm-1 to 3432 cm-1 respectively. While, the EDX study pronounced strong band at 3 keV evidenced the presence of metallic silver. The synthesized nanoparticles showed better antimicrobial activity against the standard Gram-negative bacteria such as Escherichia coli (31.25 µg/ml), Klebsiella pneumoniae (62.5 µg/ml), and Pseudomonas aeruginosa (15.6 µg/ml) respectively. In addition, the prolific MIC values against the drug resistant clinical pathogens namely Acinetobacter baumannii (7.81 µg/ml), multidrug resistant Staphylococcus aureus (7.81 µg/ml) and Proteus mirabilis (15.6 µg/ml) were its advantage. Alternatively, considerable activity towards the wound infecting bacteria Enterococcus faecium (62.5 µg/ml) was its value-added properties. In conclusion, the silver nanoparticles produced from marine Streptomyces sp. Al-Dhabi-89 exhibited potential activity against both standard and clinical drug resistant microbial pathogens.