Microbial-Physical Synthesis of Fe and Fe3O4 Magnetic Nanoparticles Using Aspergillus niger YESM1 and Supercritical Condition of Ethanol

Magnetic Fe and Fe3O4 (magnetite) nanoparticles are successfully synthesized using Aspergillus niger YESM 1 and supercritical condition of liquids. Aspergillus niger is used for decomposition of FeSO4 and FeCl3 to FeS and Fe2O3, respectively. The produced particles are exposed to supercritical condition of ethanol for 1 hour at 300∘ C and pressure of 850 psi. The phase structure and the morphology measurements yield pure iron and major Fe3O4 spherical nanoparticles with average size of 18 and 50 nm, respectively. The crystal size amounts to 9 nm for Fe and 8 nm for Fe3O4. The magnetic properties are measured to exhibit superparamagneticand ferromagnetic-like behaviors for Fe and Fe3O4 nanoparticles, respectively. The saturation magnetization amounts to 112 and 68 emu/g for Fe and Fe3O4, respectively. The obtained results open new route for using the biophysical method for large-scale production of highly magnetic nanoparticles to be used for biomedical applications

Characterization of Biofilm Forming Marine Pseudoalteromonas spp

Biofilm forming bacteria are omnipresent in the marine environment. Pseudoalteromonas is one of the largest within the γ-proteobacteria class, and a member of marine bacteria. Species of Pseudoalteromonas are generally found in association with marine eukaryotes. In this work, we present the isolation and characterization of two strains forming biofilm on rock surface and associated with marine sponge. They were identified using 16SrDNA as Pseudoalteromonas prydzensis alex, and Pseudoalteromonas sp. alex. They showed the highest titer in biofilm formation quantified using the test tube method using crystal violet

Biofilm Formation by Marine Cobetia marina alex and Pseudoalteromonas spp: Development and Detection of Quorum Sensing N-Acyl Homoserine Lactones (AHLs) Molecules

Surfaces submerged in seawater are colonized by various microorganisms, resulting in the formation of heterogenic marine biofilms. This work aims to evaluate the biofilm formation by Cobetia marina alex and doing a comparative study between this promising strain with the two bacterial strains isolated previously from the Mediterranean seawater, Alexandria, Egypt. Three strains; Cobetia marina alex, Pseudoalteromonas sp. alex, and Pseudoalteromonas prydzensis alex were screened for biofilm formation using the crystal violet (CV) quantification method in a single culture. The values of biofilm formed were OD600= 3.0, 2.7, and 2.6, respectively leading to their selection for further evaluation. However, factors affecting biofilm formation by C. marina alex were investigated. Biofilm formation was evaluated in single and multispecies consortia. Synergistic and antagonistic interactions proved in this work lead to the belief that these bacteria have the capability to produce some interesting signal molecules N-acyl Homoserine Lactones (AHLs

Agarase Production by Marine Pseudoalteromonas sp. MHS: Optimization, and Purification

Agar is an essential polysaccharide that has been utilized in numerous fields. Many kinds of literature have been published regarding agarolytic microorganisms’ isolation and agarases biochemical studies. In this search, a local marine agarolytic bacterium associated with marine alga Ulva lactuca surface was isolated and identified as Pseudoalteromonas sp. MHS. The agarase production was parallel to the growth of Pseudoalteromonas sp. MHS as cells displayed a lag phase (2 h), subsequently an exponential growth that prolonged till 10 h where maximum growth (OD550nm = 3.9) was achieved. The enzyme activity increased rapidly as cells increased exponentially where the maximum activity of 0.22 U/mL was achieved after 8h and remained constant till 12 h during the stationary phase of growth. Agarase production was optimized using Plackett-Burman statistical design by measuring enzyme activity as a response and the design was validated using a verification experiment; the activity of the enzyme increased from 0.22 U/mL to 0.29 U/mL. Pseudoalteromonas sp. MHS agarase was partially purified and its molecular weight (MW) was determined by SDSPAGE (15-25 kDa). Agarase showed approximately 94% of its activity at 40 °C. The enzyme stability decreased as the temperature increased; the enzyme could retain about 98, 90, 80, 75, and 60% of its activity at 20, 30, 40, 50, and 60 °C, respectively. Biomass of the red alga Pterocladia capillacea proved to be a suitable substrate for agarase production using Pseudoalteromonas sp. MHS; the enzyme activity recorded after 24 h of incubation was 0.35 U/mL compared to 0.29 U/mL from the optimized medium

Isolation and identification of a prodigiosin-like pigment producer Vibrio sp. isolate from a sea snail Thais sp

Marine bacteria associated with Mediterranean Sea snails have not been explored intensively. The aim of the study was to explore marine pigmented bacteria associated with the sea snail Thais sp. commonly found in Alexandria seashores where a red-pigmented bacterium was isolated and identified as Vibrio sp. based on phenotypic traits and 16S rRNA gene sequence analysis. Based on ultraviolet-visible (UV-vis) spectral and liquid chromatography-mass spectrometry (LC-MS) analysis, the red pigment was identified as a prodigiosin-like pigment. To our knowledge, this is the first report on the isolation of a Vibrio strain from a gastropod able to synthesize prodigiosin. Maximum production (7 mg/L) was achieved after two days in MZM cultures containing 0.5% mannitol and 2.5% soybean meal, pH 7 and statically incubated at 30 °C, using Plackett-Burman factorial design. RT-qPCR revealed that pks-1 and nrps responsible for the biosynthesis of prodigiosin and other bioactive compounds were up-regulated by the addition of sublethal concentrations of imipenem, DMSO, and the heavy metals copper oxide and chromium VI oxide. Through this work it can be concluded that marine invertebrates should be further explored for bacteria producing bioactive compounds whose synthesis can be enhanced or blocked depending on the carbon and nitrogen sources adopted in the fermentation medium. Moreover, the mRNA expression of the genes responsible for prodigiosin-like pigment production can be up-regulated through adopting sub-lethal concentrations of stress inducing compounds as well as co-cultivatio

Evaluation of Bio-Friendly Formulations From Siderophore-Producing Fluorescent \u3ci\u3ePseudomonas\u3c/i\u3e as Biocontrol Agents for the Management of Soil-Borne Fungi, \u3ci\u3eFusarium oxysporum\u3c/i\u3e and \u3ci\u3eRhizoctonia solani\u3c/i\u3e

Secretion of siderophores by Pseudomonas aeruginosa F2 and P. fluorescens JY3 was evaluated on chrome azurol S (CAS) agar plates and their inhibitory effect was inspected against Fusarium oxysporum and Rhizoctonia solani. Production of siderophores as biocontrol agents from F2 and JY3 was accomplished in two optimized media. Afterward, cell-free supernatants of the bacterial cultures containing siderophores were used for the preparation of two bio-friendly formulations for the management of F. oxysporum and R. solani under greenhouse conditions. The investigated bacterial isolates, F2 and JY3, showed antagonistic activity in vitro against F. oxysporum and R. solani and produced siderophores in optimized media with high efficiency. Colonies of both bacterial isolates were grown exponentially with a constant specific growth rate of 0.07 h−1 and 0.27 h−1, correspondingly. Siderophores estimated in 10 µL reached their highest value of 16.95% at 47 h and 19.5% at 48 h for isolate F2 and JY3, respectively. Formulations of siderophore-generating F2 and JY3 reduced damping-off caused by F. oxysporum by 40% and 80%, while the reduction percentage of damping-off caused by R. solani reached 87.5% and 62.5%, correspondingly. Moreover, both formulations encouraged the growing of wheat plants where the fresh and dry weight of shoots and roots were increased compared to the treatment with each fungus. In conclusion, bio-friendly formulations resulting from this investigation can play an active role in managing soil-borne diseases

Potential Egyptian bacterial consortium for oil spill treatment: A laboratory simulation

The purpose of the study was to reach high efficiency of an oil-degrading bacteria to be used in oil-spill treatment separately or with bacterial consortium. The consortium effect on crude oil was examined by GC-MS. The consortium degraded 99.2% of crude oil after 7 days, while Enterobacter sp. ASH, as individual culture, degraded 80% only. This isolate was identified based on 16S rRNA gene sequence analysis, morphological, physiological, and biochemical characterization. ANOVA analysis showed that medium volume and crude oil concentration are the significant factors. The results showed that the mixed consortium showed better biodegradation abilities than E. sp. ASH

A De Novo Optimized Cell-Free System for the Expression of Soluble and Active Human Tumor Necrosis Factor-Alpha

Cell-free (in vitro) expression is a robust alternative platform to the cell-based (in vivo) system for recombinant protein production. Tumor necrosis factor-alpha (TNF-α) is an effective pro-inflammatory cytokine with pleiotropic effects. The aim of the current study was de novo optimized expression of soluble and active human TNF-α by an in vitro method in an E. coli-based cell-free protein synthesis (CFPS) system and its biological activity evaluation. The codon-optimized synthetic human TNF-α gene was constructed by a two-step PCR, cloned into pET101/D-TOPO vector and then expressed by the E. coli CFPS system. Cell-free expression of the soluble protein was optimized using a response surface methodology (RSM). The anticancer activity of purified human TNF-α was assessed against three human cancer cell lines: Caco-2, HepG-2 and MCF-7. Data from RSM revealed that the lowest value (7.2 µg/mL) of cell-free production of recombinant human TNF-α (rhTNF-α) was obtained at a certain incubation time (6 h) and incubation temperature (20 °C), while the highest value (350 µg/mL) was recorded at 4 h and 35 °C. This rhTNF-α showed a significant anticancer potency. Our findings suggest a cell-free expression system as an alternative platform for producing soluble and functionally active recombinant TNF-α for further research and clinical trials

Statistical Optimization and Characterization of Prodigiosin from a Marine Serratia rubidaea RAM-Alex

This research sought to determine optimal conditions to maximize prodigiosin production by an indigenous Egyptian marine bacterial strain Serratia rubidaea RAM_Alex. Serratia rubidaea RAM_Alex isolated from bivalve samples of Temsah Lake, Ismailia, Egypt was used to investigate the production of the natural red pigment prodigiosin. Pigment production was assayed in different growth conditions using Nutrient broth as production medium. The water insoluble red pigment was extracted using ethanol and further purified by organic solvents. The pigment extract showed absorbance with a UV-Vis spectrophotometer at 535 nm and further characterized using TLC, FTIR and 1H-NMR. A statistical screening procedure was adopted to select the main factors affecting production. Analyses of Plackett- Burman design results demonstrated that peptone, NaCl, and culture volume were the most important independent variables. The near optimum medium contained (g/L): peptone 7, beef extract 5, yeast extract 1, NaCl 10, pH 6, using 25 ml culture volume, 100 ¼l inoculum size and incubation statically for 48 h at 30oC. When this condition was employed, a two fold increase in pigment yield was achieved reaching ~1600.511 mg/l

Production and statistical optimization of cholesterol-oxidase generated by Streptomyces sp. AN strain

Abstract Background Cholesterol oxidases (CHOs) have attracted enormous attention because of their wide biotechnological potential. The present study explores the production of CHOs by Streptomyces sp. AN. Evaluation of culture conditions affecting enzyme production, medium optimization and released metabolite characteristics were also investigated. Results The current work reports the isolation of 37 colonies (bacteria/actinobacteria) with different morphotypes from different soil/water samples. The isolate-coded AN was selected for its high potency for CHO production. Morphological characteristics and the obtained partial sequence of 16srRNA of AN showed 99.38% identity to Streptomyces sp. strain P12–37. Factors affecting CHO production were evaluated using Plackett-Burman (PB) and Box-Behnken (BB) statistical designs to find out the optimum level of the most effective variables, namely, pH, starch, NH4NO3 and FeSO4.7H2O with a predicted activity of 6.56 U/mL. According to this optimization, the following medium composition was considered to be optimum (g/L): cholesterol 1, starch 6, MgSO4.7H2O 0.1, CaCl2 0.01, FeSO4.7H2O 0.1, NH4NO3 23.97, yeast extract (YE) 0.2, K2HPO4 0.01, KH2PO4 0.1, NaCl 0.01, Tween 20 0.01, pH 6.36 and incubation temperature (30 °C) for 9 days. Spectophotometric analysis for released metabolites against cholesterol (standard) via Fourier-transform infrared spectroscopy (FTIR) and differential scanning calorimetry (DSC) was carried out. FTIR spectrum showed the appearance of new absorption peaks at 1644 and 1725cm−1; this confirmed the presence of the Keto group (C=O) stretch bond. Besides, fermentation caused changes in thermal properties such as melting temperature peak (99.26; 148.77 °C), heat flow (− 8; − 3.6 Mw/mg), capacity (− 924.69; − 209.77 mJ) and heat enthalpy (− 385.29; 69.83 J/g) by comparison to the standard cholesterol as recognized through DSC thermogram. These changes are attributed to the action of the CHO enzyme and the release of keto derivatives of cholesterol with different properties. Conclusion Streptomyces sp. AN was endowed with the capability to produce CHO. Enzyme maximization was followed using a statistical experimental approach, leading to a 2.6-fold increase in the overall activity compared to the basal condition. CHO catalyzed the oxidation of cholesterol; this was verified by the appearance of a new keto group (C=O) peak at 1644 and 1725 cm−1 observed by FTIR spectroscopic analysis. Also, DSC thermogram demonstrates the alteration of cholesterol triggered by CHO