Maximization of red pigment production from Streptomyces sp. LS1 structure elucidation and application as antimicrobial/antifouling against human pathogens and marine microbes.

BACKGROUND: Natural dyes are present in living organisms such as animals and plants and microorganisms such as fungi, bacteria, algae, and yeast. Pigments are fast and easy growth by using cheap components and do not effect by environmental conditions because they required some physical factors like heat, light, and pH and also they have many biotechnological applications such as medical and industrial needs. The natural pigments can act as antimicrobial agents and are used in drug manufacturing. Also, it can be used in the food industry as natural colorants instead of the synthetic colorants due to their safety on human health and low toxicity when emitted into the environment. RESULTS: A pigmented actinomycetes LS1 strain isolated from El Mahmoudia canal (sediment soil) located in Egypt was microscopically examined and identified as Streptomyces sp. by molecular approach. Extraction, purification, and characterization of produced red pigment metabolite like carotenoids related were established based on spectroscopic studies and comparing the data from the literature. Factors (nutritional and physical) influencing red pigmentation by this isolate were investigated through One Variable At Time (OVAT), and then, the optimal levels of the significant key variables were recorded. Also, the productivity yield reached 30 mg of dried purified pigment/gram dry weight. The biological activity of the red product was tested against Gram-positive and Gram-negative marine bacterial pathogens; the recorded antimicrobial activity is more prominent against (P. aeruginosa ATCC 9027, K. pneumoniae ATCC 13883, S. aureus ATCC 6538, B. subtilis ATCC 6633 and E. coli ATCC 10418) at nearly 0.07 mg mL-1 concentration. Also, the tested red pigment showed a positive antifouling activity (AF) against marine microbes; the activity increased by increasing the pigment concentrations from 1 to 3 mg mL-1. CONCLUSION: The present work focused on the optimization of culture conditions for the production of red pigment by Streptomyces sp. LS1; then, the antibacterial activity and antifouling activity of the produced pigments were tested.

Various In Vitro Bioactivities of Secondary Metabolites Isolated from the Sponge Hyrtios aff. Erectus from the Red Sea Coast of Egypt.

OBJECTIVES: The present study revealed the presence of bioactive constituents in Hyrtios aff. erectus sponge (HES) extract collected from the Red Sea using skin and scuba diving. MATERIALS AND METHODS: Cytotoxicity was tested against hepatocellular carcinoma cell lines as a prescreening test. RESULTS: The HES extract had high contents of total phenolic compounds (0.061 mg/g), flavonoids (0.2839 mg/g), and carotenoids (1.976 mg/g). Moreover, the HES extract showed high antioxidant capacity with 93.0% and 99% at 1 mg using 2.2'-Diphenyl-α-picrylhydrazyl and 2.2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid), respectively. Cytotoxic activity against cancerous cell lines showed that the HES extract could inhibit cell growth effectively with IC50=47.5 µg/mL. Furthermore, anticancer activity using protein tyrosine kinase and sphingosine kinase 1 inhibitor screening assays resulted in 71.66% and 85.21% inhibition activity, respectively. The anti-inflammatory assays showed that the inhibition activity against cyclooxygenase (COX1), COX2, interleukin-6, and tumor necrosis factor-α was 71.82%, 81.13%, 80.89%, and 59.74%, respectively. At the same time, the anti-Alzheimer results using acetylcholine inhibition assay showed high activity at 1 mg with 83.51%. Additionally, the antiviral activity using the reverse transcriptase inhibition assay was 91.70%. CONCLUSION: This marine sponge isolated from the Red Sea showed tremendous activity against many diseases and it is considered an excellent source for bioactive pharmaceutical compounds.

Marine Algae of the Genus Gracilaria as Multi Products Source for Different Biotechnological and Medical Applications.

BACKGROUND: Gracilaria has been shown to be an important source of marine bioactive natural biomaterials and compounds. Although there are no enough patents used Gracilaria worldwide, the current study tries to put the Gracilaria on the spot for further important patents in the future. OBJECTIVE: The current study investigates the pharmaceuticals and biochemical activity of Gracilaria because no previous studies have been carried out to examine the biochemical and pharmaceutical activates of Gracilaria from the Suez Canal of Egypt as an excellent source for bioactive compounds. METHODS: Different advanced experimental models and analytical techniques, such as cytotoxicity, total antioxidant capacity, anticancer, and anti-inflammatory profiling were applied. The phytochemical analysis of different constituents was also carried out. RESULTS: The mineral analysis revealed the presence of copper (188.3 ppm) and iron (10.07 ppm) in addition to a remarkable wealth of selenium and sulfur contents giving up to 36% of its dry mass. The elemental analysis showed high contents of sulfur and nitrogen compounds. The GCMS profiling showed varieties of different bioactive compounds, such as fatty acids, different types of carotenoids in addition to pigments, alkaloids, steroids. Many other compounds, such as carbohydrates and amino acids having antioxidant, anti-inflammatory, and antiviral activities, etc. were identified. The cytotoxicity activity of Gracilaria marine extract was very effective against cancerous cell lines and showed high ability as a potent antitumor due to their bioactive constituents. Specialized screening assays using two anticancer experimental models, i.e., PTK and SKH1 revealed 77.88% and 84.50% inhibition anticancer activity; respectively. The anti-inflammatory activities investigated using four different experimental models, i.e., COX1, COX2, IL6, and TNF resulted in 68%, 81.76%, 56.02% and 78.43% inhibition; respectively. Moreover, Gracilaria extracts showed potent anti-Alzheimer with all concentrations. CONCLUSION: Gracilaria proved to be a multi-product source of marine natural products for different biotechnological applications. Our recommendation is to investigate the Gracilaria bioactive secondary metabolites in order to create and innovate in more patents from current important seaweeds (Gracilaria).

Pesudomance sp. Bacteria Associated with Marine Sponge as a Promising and Sustainable Source of Bioactive Molecules.

BACKGROUND: The study was conducted to identify the bacterial strain associated with marine sponge Hyrtiosaff. erectus collected from the Red Sea coastal water and to assess the utilization of their secondary metabolites for human benefit as antioxidant, anti-Alzheimer, anti-viral, anticancer and anti-inflammatory agent. METHODS: After biochemical identification of Pesudomance sp. bacterial strain, the total polyphenol contents, cytotoxic, antioxidant, anti-Alzheimer, anti-viral, anticancer and anti-inflammatory activity of the Pesudomance sp. ethyl acetate extract were investigated by applying different biochemical assays. Polyphenol contents were investigated using spectrophotometric techniques. Antioxidant activity was determined by 1,1-diphenyl-2-picrylhydrazyl radical (DPPH), and 2,2/-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) ABTS radical scavenging activity assays. The cytotoxic effects were investigated by using the human cancerous cell lines. RESULTS: The anti-Alzheimer, anti-viral, anticancer and anti-inflammatory activities were determined using ELISA. Qualitative phytochemical analysis of the Pesudomance sp. extract demonstrated the presence of a large and diverse group of substances such as alkaloids, carbohydrates, flavonoids, phenols, terpenoids, saponins, and tannins. The strong antioxidant activity of the Pesudomance sp. extract was mainly attributed to the protective role of polyphenols against reactive oxygen. It was also observed that Pesudomance sp. extract possessed significant anti-Alzheimer activity with 94% at 1 mg. The extract showed also high antiviral activity (90%) using reverse transcriptase enzymes inhibition assay. The examination of the anticancer activity by applying two experimental models, i.e., PTK and SHKI cleared out high significant percentages of 76.19 and 83.09 %; respectively. CONCLUSION: The anti-inflammatory profiling using TNF, COX1, COX2, IL6 also revealed high antiinflammatory activity with different metabolic pathway of 62.70, 75.444, 79.27 and 54.15 %; respectively. The present study concluded that ethyl acetate extract of Pesudomance sp. possessed strong antioxidant, anti-Alzheimer, and anti-viral, anticancer and anti-inflammatory activities. Further studies are required to purify the bioactive compounds.

Hydrogel film loaded with new formula from manuka honey for treatment of chronic wound infections.

OBJECTIVES: Chronic wound infections can be effectively treated using hydrogel loaded with an extract from manuka honey. METHODS: This study was performed on bacterial isolates from patients with infected wounds at Alexandria Main University Hospital (Alexandria, Egypt). Isolates were exposed to hydrogel sheets composed of chitosan and gelatin and loaded with a new formula from manuka honey. RESULTS: The results illustrate the antibacterial activity of the formula extracted from manuka honey against Staphylococcus aureus, Streptococcus pyogenes, Acinetobacter baumannii, Pseudomonas aeruginosa and Proteus mirabilis. Screening of the hydrogel by electron microscopy showed the ultrastructure of the gel. CONCLUSIONS: A hydrogel sheet composed of chitosan and gelatin loaded with a new formula extracted from manuka honey can be used as a dressing for chronic infected wounds.

Protective role of antioxidants capacity of Hyrtios aff. Erectus sponge extract against mixture of persistent organic pollutants (POPs)-induced hepatic toxicity in mice liver: biomarkers and ultrastructural study.

The current study investigates the hepatoprotective effect of Hyrtios aff. Erectus sponge extract against POPs intoxication on endogenous antioxidant enzymes and lipid peroxidation in mice liver tissue. In the present study, the mice BALB/C were assigned into four groups: group I: received saline subcutaneously for 7 days and served as negative control; group II: received subcutaneously for 7 days, 130.6 mg/100 g/b. w/day POPs mixture(mixture of PCB 28, PCB 52,, PCB 101, PCB 118, PCB 153, PCB 138 and PCB 180, alpha-Hexachlorocyclohexane, beta-Hexachloro-cyclohexane, gamma-hexachlorocyclohexane, Aldrin, O,P'-DDE, Dieldrin, P,p DDE, O,P DDD, Endrin, P,p DDD and P,pDDT were extracted from sediments collected from Lake Mariout), and served as induced group; group III: pretreated with Hyrtios aff. Erectus sponge extract for 7 days, as a protection dose and then treated with POPs as group II and served as protective group; and group IV: received i.p Hyrtios aff. Erectus sponge extract of dose 0.7 mg/100 g b.wt/day for 7 days and served as positive control. After 7 days (experimental period), mice were scarified and the liver was harvested for biochemical estimation. Significant reduction in lipid peroxidation (p < 0.002) was noticed compared to POPs-protected group. The antioxidant biomarkers levels were significantly increase as the hepatic GSH and GST increased by 69.9 and 89.9%, respectively. Such increase was accompanied by a decrease in tyrosine kinase activity by 59.82%, additionally remarkable histopathological changes in liver tissue indicate the protective effect of Hyrtios aff. Erectus sponge extract. The results of this study revealed that the Hyrtios aff. Erectus sponge extract has the potential to diminish the destructive effect of POPs intoxication through enhancement of the endogenous antioxidant status. The hepatoprotective activity of Hyrtios aff. Erectus sponge extract is mediated, by the antioxidant effect of its active constituents. The active constituents of Hyrtios aff. Erectus sponge extract were identified by LC-MS/MS.

Isolation, characterization, optimization, immobilization and batch fermentation of bioflocculant produced by Bacillus aryabhattai strain PSK1.

Among others, isolate PSK1 was selected and identified by 16 S rDNA sequencing as Bacillus aryabhattai. Growth optimization of PSK1 and physicochemical parameters affected bioflocculant production was carried out by Plackett-Burman design and resulted in increasing in the activity by 4.5%. Bioflocculant production by entrapped and adsorbed immobilized microbial cells was performed using different techniques and revealed enhancement in the activity in particular with pumice adsorption. HPLC analysis of sugars and amino acids composition, FTIR and the effect of different factors on the purified PSK1 biopolymer such as presence of cations, thermal stability, pH range and clay concentration was carried out. Scanning electron microscopy (SEM) of free, immobilized cells, PSK1 bioflocculant and formed flocs were performed. The results revealed that bioflocculant PSK1 is mainly glycoprotein consists of glucose and rhamnose with a large number of amino acids in which arginine and phenylalanine were the major. SEM analysis demonstrated that PSK1 have a clear crystalline rod shaped structure. FTIR spectrum reported the presence of hydroxyl and amino groups which are preferred in flocculation process. PSK1 was soluble in water and insoluble in all other tested organic solvents, while it was thermally stable from 40 to 80 °C. Among examined cations, CaCl2 was the best coagulant. The maximum flocculation activity of the PSK1 recorded at 50 °C (92.8%), pH 2.0 (94.56%) with clay concentration range 5-9 g/l. To obtain a large amount of PSK1 bioflocculant with high flocculating activity, batch fermentation was employed. The results recorded ∼6 g/l yield after 24 h of fermentation.

Solid-state fermentation for the production of meroparamycin by streptomyces sp. strain MAR01.

The antibiotic meroparamycin was produced in the free culture system of Streptomyces sp. strain MAR01. Five solid substrates (rice, wheat bran, Quaker, bread, and ground corn) were screened for their ability to support meroparamycin production in solid-state fermentation. In batch culture, wheat bran recorded the highest antibacterial activity with the lowest residual substrate values. The highest residual substrate values were recorded for both ground corn and Quaker. On the other hand, no antibacterial activity was detected for rice as a solid substrate. The use of the original strength of starch-nitrate medium in the solid-state fermentation gave a lower antibacterial activity compared with the free culture system. Doubling the strength of this medium resulted in the increase in the activity to be equivalent to the free culture. The initial pH (7.0) of the culture medium and 2 ml of spore suspension (1 ml contains 5x10(9) spores/ml) were the optima for antibiotic production. The water was the best eluent for the extraction of the antibiotic from the solid-state culture. Ten min was enough time to extract the antibiotic using a mixer, whereas, 60 min was required when shaking was applied. Semicontinuous production of meroparamycin using a percolation method demonstrated a more or less constant antibacterial activity over 4 runs (450-480 microg/ml). The semicontinuous production of the antibiotic was monitored in a fixed-bed bioreactor and the maximum activity was attained after the fourth run (510 microg/ml) and the overall process continued for 85 days.

Meroparamycin production by newly isolated Streptomyces sp. strain MAR01: taxonomy, fermentation, purification and structural elucidation.

Twelve actinomycete strains were isolated from Egyptian soil. The isolated actinomycete strains were then screened with regard to their potential to generate antibiotics. The most potent of the producer strains was selected and identified. The cultural and physiological characteristics of the strain identified the strain as a member of the genus Streptomyces. The nucleotide sequence of the 16S rRNA gene (1.5 kb) of the most potent strain evidenced a 99% similarity with Streptomyces spp. and S. aureofaciens 16S rRNA genes, and the isolated strain was ultimately identified as Streptomyces sp. MAR01. The extraction of the fermentation broth of this strain resulted in the isolation of one major compound, which was active in vitro against gram-positive, gram-negative representatives and Candida albicans. The chemical structure of this bioactive compound was elucidated based on the spectroscopic data obtained from the application of MS, IR, UV, 1H NMR, 13C NMR, and elemental analysis techniques. Via comparison to the reference data in the relevant literature and in the database search, this antibiotic, which had a molecular formula of C19H29NO2 and a molecular weight of 303.44, was determined to differ from those produced by this genus as well as the available known antibiotics. Therefore, this antibiotic was designated Meroparamycin.