Simultaneous polyhydroxyalkanoates and rhamnolipids production by Thermus thermophilus HB8

The ability of Thermus thermophilus HB8 to produce simultaneously two environmentally-friendly biodegradable products, polyhydroxyalkanoates (PHAs) and rhamnolipids (RLs), using either sodium gluconate or glucose as sole carbon source, was demonstrated. The utilization of sodium gluconate resulted in higher levels of PHAs and RLs production than when glucose was used as sole carbon source. The initial phosphate concentration (as PO43-) influences both PHAs and RLs productions that were increased during cultivation time. PHAs accumulation was enhanced (> 300 mg/L) after 72 h of cultivation in an initial [PO43-] of 25 mM, while RLs production (> 200 mg/L) was started after 35 h and continued until 72 h of cultivation, in a phosphate-limited medium containing initially 5 mM of [PO43-]. In addition, the combine effect of initial [PO43-] and cultivation time on biomass, PHAs and RLs production was evaluated from 2D contour plots. The results revealed that low initial phosphate concentrations (up to 5 mM) and long incubation time (72 h) promoted RLs biosynthesis while higher initial phosphate concentrations (up to 25 mM) where favorable for biomass and PHAs production. The molecular composition of the produced bio-products was identified. The accumulated PHAs were co-polymers which mainly consisted of 3-hydroxydecanoate (3HD) as resulted by gas chromatography (GC) analysis. The secreted RLs were extracted and their total mixture contained both mono- and di- RLs identified by thin-layer chromatography (TLC). Moreover, the molecular composition of the produced RLs characterized in details by LC-MS analysis showed a plethora of diversity including mono-, and di-RLs, di-rhamno-monolipidic congeners differing in the length of the lipidic chain, which additionally were found to be saturated or unsaturated in some cases

Thermus thermophilus genome analysis: benefits and implications

The genome sequence analysis of Thermus thermophilus HB27, a microorganism with high biotechnological potential, has recently been published. In that report, the chromosomal and the megaplasmid sequence were compared to those of other organisms and discussed on the basis of their physiological and metabolic features. Out of the 2,218 putative genes identified through the large genome sequencing project, a significant number has potential interest for biotechnology. The present communication will discuss the accumulating information on molecules participating in fundamental biological processes or having potential biotechnological importance

Synthesis and antimicrobial evaluation of a pyrazoline-pyridine silver(I) complex: DNA-interaction and anti-biofilm activity

The emergence of resistant bacterial strains mainly due to misuse of antibiotics has seriously affected our ability to treat bacterial illness, and the development of new classes of potent antimicrobial agents is desperately needed. In this study, we report the efficient synthesis of a new pyrazoline-pyridine containing ligand L1 which acts as an NN-donor for the formation of a novel silver (I) complex 2. The free ligand did not show antibacterial activity. High potency was exhibited by the complex against three Gram-negative bacteria, namely Escherichia coli, Pseudomonas aeruginosa and Acinetobacter baumanii with the minimum inhibitory concentration (MIC) ranging between 4 and 16 μg/mL (4.2–16.7 μM), and excellent activity against the fungi Candida albicans and Cryptococcus neoformans (MIC ≤ 0.25 μg/mL = 0.26 μM). Moreover, no hemolytic activity within the tested concentration range was observed. In addition to the planktonic growth inhibition, the biofilm formation of both Methicillin-resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa was significantly reduced by the complex at MIC concentrations in a dose-dependent manner for Pseudomonas aeruginosa, whereas a biphasic response was obtained for MRSA showing that the sub-MIC doses enhanced biofilm formation before its reduction at higher concentration. Finally, complex 2 exhibited strong DNA binding with a large drop in DNA viscosity indicating the absence of classical intercalation and suggesting the participation of the silver ion in DNA binding which may be related to its antibacterial activity. Taken together, the current results reveal that the pyrazoline-pyridine silver complexes are of high interest as novel antibacterial agents, justifying further in vitro and in vivo investigation

The Effect of Fused 12-Membered Nickel Metallacrowns on DNA and their Antibacterial Activity

The synthesis, characterization and the biological study of a series of Ni(ll)2(carboxylato)2 [12- MCNi(II)N(shi)2(pko)2-4][12-MCNi(ii)N(sh03(pko)-4] (CH3OH)3(H3O) fused 12-membered metallacrowns with 10 metal ions and commercial available herbicides or anti-inflammatory drugs as carboxylato ligands are reported. All the compounds have a mixed ligand composition with salicylhydroxamic acid and di-2-pyridylketonoxime as chelate agents. The compounds construct metallacrown cores {[12-MCNi(n)N(sj02(pko)2-4][12-MCNi(ll)N(shO3(pko)-4]}2+ following the pattern [-Ni-O-N-]4. The neutral decanuclear [Ni(II)(A)]2[12-MCNi(II)N(shi)2(pko)2-4][12-MCNi(II)N(pko)3(pko)-4] fused metallacrown, consists of two [12-MCM(ox)N(ligand)-4] units the {Ni(ll)(A)[12-MCNi(II)N(shi)2(pko)2-4]} and {Ni(II)(A)[12-MCNi(II)N(shi)3(pko)-4]} with 1+ and 1- charge, respectively. Each metallacrown unit has four ring Ni(II) ions and one additional encapsulated Ni(II) ion in planar arrangement. The anionic unit is bonded with cationic one creating binuclear moieties. The herbicide or antiiflammatory carboxylato ligands are bridging the central octahedral nickel atom with a ring metal ion in a bindetate fashion. The effect on DNA and their antibacterial activity was examined. The changes in the mobility can be attributed to the altered structures of the pDNA treated with Ni(II) complexes. Evaluating the data of the antibacterial activity of the compounds tested, we can conclude that nickel complexes present strong antibacterial activity

Elucidation of One Step Synthesis of PEGylated CuFe Bimetallic Nanoparticles. Antimicrobial Activity of CuFe@PEG vs Cu@PEG

There is a growing field of research on the physicochemical properties of bimetallic nanoparticles (BMNPs) and their potential use in different applications. Meanwhile, their antimicrobial activity is scarcely reported, although BMNPs can potentially achieve unique chemical transformations and synergetic effects can be presented. Towards this direction a reproducible simple hybrid polyol process under moderate temperature solvothermal conditions has been applied for the isolation of non-oxide contaminated bimetallic CuFe nanoparticles (NPs). 1,2-propylene glycol (PG), tetraethylene glycol (TEG) and polyethylene glycol (PEG 8000), that exhibit different physicochemical properties, have been utilized to regulate the size, structure, composition and the surface chemistry of NPs. The BMNPs were found to be of small crystalline size, 30–45 nm, and high hydrophilicity, different wt% percentage of organic coating and variable hydrodynamic size and surface charge. The antimicrobial activity of the BMNPs was evaluated against the bacterial strains B. subtilis, E. coli and fungus S. cerevisiae. The IC50 values for CuFe NPs were found significantly lower compared with Cu NPs of the same size, revealing an enhancement in the antimicrobial activity when iron and copper coexist in the crystal structure. The reactive oxygen species (ROS) production was measured intracellularly and extracellularly by the nitroblue tetrazolium assay in the fungal cultures. No extracellular ROS were measured suggesting that both CuFe and Cu NPs enter the fungal cells during the incubation, also verified by optical imaging of the fungal cells in the presence of NPs. Higher ROS concentrations were generated intracellularly for CuFe NPs supporting different red/ox reaction mechanisms

Synthesis and biological evaluation of PEGylated CuO nanoparticles

There is a growing field of research into the physicochemical properties of metal oxide nanoparticles (NPs) and their potential use against tumor formation, development and progression. Coated NPs with biocompatible surfactants can be incorporated into the natural metabolic pathway of the body and specifically favor delivery to the targeted cancerous cells versus normal cells. Polyethylene glycol (PEG) is an FDA approved, biocompatible synthetic polymer and PEGylated NPs are regarded as “stealth” nanoparticles, which are not recognized by the immune system. Herein, PEGylated cupric oxide nanoparticles (CuO NPs) with either PEG 1000 or PEG 8000 were hydrothermally prepared upon properly adjusting the reaction conditions. Depending on the reaction time CuO NPs in the range of core sizes 11–20 nm were formed, while hydrodynamic sizes substantially varied (330–1120 nm) with improved colloidal stability in PBS. The anticancer activity of the NPs was evaluated on human cervical carcinoma HeLa cells by using human immortalized embryonic kidney 293 FT cells as a control. Viability assays (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, MTT) revealed that CuO NPs could selectively reduce viability of tumor cells (IC50 values 11.91–25.78 μg/mL). Reactive oxygen species (ROS) production, cell membrane damage and apoptotic DNA laddering were also evident by nitroblue tetrazolium (NBT) reduction, lactate dehydrogenase (LDH) release assays and DNA electrophoresis, respectively. CuO NPs strongly inhibited lipoxygenase (LOX) enzymatic activity with IC50 values 4–5.9 μg/mL, highlighting in that manner their anti-inflammatory activity

In-depth synthetic, physicochemical and in vitro biological investigation of a new ternary V(IV) antioxidant material based on curcumin.

Curcumin is a natural product with a broad spectrum of beneficial properties relating to pharmaceutical applications, extending from traditional remedies to modern cosmetics. The biological activity of such pigments, however, is limited by their solubility and bioavailability, thereby necessitating new ways of achieving optimal tissue cellular response and efficacy as drugs. Metal ion complexation provides a significant route toward improvement of curcumin stability and biological activity, with vanadium being a representative such metal ion, amply encountered in biological systems and exhibiting exogenous bioactivity through potential pharmaceuticals. Driven by the need to optimally increase curcumin bioavailability and bioactivity through complexation, synthetic efforts were launched to seek out stable species, ultimately leading to the synthesis and isolation of a new ternary V(IV)-curcumin-(2,2’-bipyridine) complex. Physicochemical characterization (elemental analysis, FT-IR, Thermogravimetry (TGA), UV-Visible, NMR, ESI-MS, Fluorescence, X-rays) portrayed the solid-state and solution properties of the ternary complex. Pulsed-EPR spectroscopy, in frozen solutions, suggested the presence of two species, cis- and trans-conformers. Density Functional Theory (DFT) calculations revealed the salient features and energetics of the two conformers, thereby complementing EPR spectroscopy. The well-described profile of the vanadium species led to its in vitro biological investigation involving toxicity, cell metabolism inhibition in S. cerevisiae cultures, Reactive Oxygen Species (ROS)-suppressing capacity, lipid peroxidation, and plasmid DNA degradation. A multitude of bio-assays and methodologies, in comparison to free curcumin, showed that it exhibits its antioxidant potential in a concentration-dependent fashion, thereby formulating a bioreactivity profile supporting development of new efficient vanado-pharmaceuticals, targeting (extra)intra-cellular processes under (patho)physiological conditions

DING Proteins from Phylogenetically Different Species Share High Degrees of Sequence and Structure Homology and Block Transcription of HIV-1 LTR Promoter

Independent research groups reported that DING protein homologues isolated from bacterial, plant and human cells demonstrate the anti-HIV-1 activity. This might indicate that diverse organisms utilize a DING-mediated broad-range protective innate immunity response to pathogen invasion, and that this mechanism is effective also against HIV-1. We performed structural analyses and evaluated the anti-HIV-1 activity for four DING protein homologues isolated from different species. Our data show that bacterial PfluDING, plant p38SJ (pDING), human phosphate binding protein (HPBP) and human extracellular DING from CD4 T cells (X-DING-CD4) share high degrees of structure and sequence homology. According to earlier reports on the anti-HIV-1 activity of pDING and X-DING-CD4, other members of this protein family from bacteria and humans were able to block transcription of HIV-1 and replication of virus in cell based assays. The efficacy studies for DING-mediated HIV-1 LTR and HIV-1 replication blocking activity showed that the LTR transcription inhibitory concentration 50 (IC50) values ranged from 0.052–0.449 ng/ml; and the HIV-1 replication IC50 values ranged from 0.075–0.311 ng/ml. Treatment of cells with DING protein alters the interaction between p65-NF-κB and HIV-1 LTR. Our data suggest that DING proteins may be part of an innate immunity defense against pathogen invasion; the conserved structure and activity makes them appealing candidates for development of a novel therapeutics targeting HIV-1 transcription