The biosynthetic gene cluster for the aromatic polyketide steffimycin of the anthracycline family has been cloned and characterized from “ Streptomyces steffisburgensis ” NRRL 3193. Sequence analysis of a 42.8-kbp DNA region revealed the presence of 36 open reading frames (ORFs) (one of them incomplete), 24 of which, spanning 26.5 kb, are probably involved in steffimycin biosynthesis. They code for all the activities required for polyketide biosynthesis, tailoring, regulation, and resistance but show no evidence of genes involved in l -rhamnose biosynthesis. The involvement of the cluster in steffimycin biosynthesis was confirmed by expression of a region of about 15 kb containing 15 ORFS, 11 of them forming part of the cluster, in the heterologous host Streptomyces albus , allowing the isolation of a biosynthetic intermediate. In addition, the expression in S. albus of the entire cluster, contained in a region of 34.8 kb, combined with the expression of plasmid pRHAM, directing the biosynthesis of l -rhamnose, led to the production of steffimycin. Inactivation of the stfX gene, coding for a putative cyclase, revealed that this enzymatic activity participates in the cyclization of the fourth ring, making the final steps in the biosynthesis of the steffimycin aglycon similar to those in the biosynthesis of jadomycin or rabelomycin. Inactivation of the stfG gene, coding for a putative glycosyltransferase involved in the attachment of l -rhamnose, allowed the production of a new compound corresponding to the steffimycin aglycon compound also observed in S. albus upon expression of the entire cluster.
Biogenic copper nanoparticles (Cu NPs) were synthesized using the aqueous crude extract of mangrove leaves,Avicennia marina(CE). GC-MS metabolite profiling of CE showed that their carbohydrates are mainly composed of D-mannose (29.21%), D-fructose, (18.51%), L-sorbose (12.91%), D-galactose (5.47%) and D-Talose (5.21%). Ultra-fine nanoparticles of 11.60 ±4.65 nm comprising Cu2O and Cu(OH)2species were obtained with a carbohydrate and phenolic content of 35.6±3.2% and 3.13±0.05 mgGA/g, respectively. The impact of the biogenic Cu NPs on wheat seedling growth was dose-dependent. Upon treatment with 0.06 mg/mL of Cu NPs, the growth was promoted by 172.78 ± 23.11 and 215.94 ± 37.76% for wheat root and shoot, respectively. However, the lowest relative growth % of 81.94 ± 11.70 and 72.46 ± 18.78% were recorded for wheat root and shoot, respectively when applying 0.43 mg/mL of Cu NPs. At this concentration, peroxidase activity (POX) of the germinated wheat seeds also decreased, while ascorbic acid oxidase (AAO) and polyphenol oxidase (PPO) activities increased. Higher uptake of copper was observed in the root relative to the shoot implying the accumulation of the nanoparticles in the former. The uptake was also higher than that of the commercial Cu NPs, which showed an insignificant effect on the seedling growth. By treating the wheat leaves in foliar application with 0.06 mg/mL of Cu NPs, their contents of Chlorophyll a, Chlorophyll b, and total chlorophyll were enhanced after 21 days of application. Meanwhile, the high concentration (0.43 mg/mL) of Cu NPs was the most effective in reducing the leaf content of chlorophyll (a, b, and total) after the same time of application. The findings of this study manifest the potential of utilizing controlled doses of the prepared biogenic Cu NPs for inhibition or stimulation of seedling growth.
ObjectiveTo isolate and evaluate the cytotoxic activity of different actinomycetes species isolated from the Red Sea coast in Sharm el-Sheikh, Egypt.MethodsForty actinomycetes strains were isolated from different sediments and seawater samples collected from the Red Sea coast in Egypt. Actinomycetes were recognized by morphological and microscopic examinations. Cell viability and cytotoxicity induced by the crude extracts on breast cancer cell lines MDA-MB-231 were assessed using methylene blue assay. The strains with promising cytotoxic activity were identified by sequencing and amplifying the 16S rRNA genes. The antibacterial activities of the crude extracts were performed using Kirby–Bauer disc diffusion method.ResultsThe results indicated that five ethyl acetate extracts exhibited cytotoxicity towards breast cancer cell lines MDA-MB-231. The highest cytotoxic activity was found for the ethyl acetate extracts of EGY2 and EGY39. The isolate EGY3 was identified as a new Streptomyces species, while the actinomycete EGY22 was found to be a member of the genus Nocardiopsis sp. The crude extract of the isolate EGY8 showed slightly high antimicrobial activity against different test microorganisms.ConclusionsThe results of the present study reveal that marine sediments of the Red Sea are a potent source of novel species of actinomycetes. The isolates may be useful in discovery of novel bioactive compounds and an important step in the development of microbial natural product research