Methylotrophic extremophilic yeast Trichosporon sp. : a soil-derived isolate with potential applications in environmental biotechnology

A yeast isolate revealing unique enzymatic activities and substrate-dependent polymorphism was obtained from autochthonous microflora of soil heavily polluted with oily slurries. By means of standard yeast identification procedures the strain was identified as Trichosporon cutaneum. Further molecular PCR product analyses of ribosomal DNA confirmed the identity of the isolate with the genus Trichosporon. As it grew on methanol as a sole carbon source, the strain appeared to be methylotrophic. Furthermore, it was also able to utilize formaldehyde. A multi-substrate growth potential was shown with several other carbon sources: glucose, glycerol, ethanol as well as petroleum derivatives and phenol. Optimum growth temperature was determined at 25°C, and strong inhibition of growth at 37°C together with the original soil habitat indicated lack of pathogenicity in warm-blooded animals and humans. The unusually high tolerance to xenobiotics such as diesel oil (>30 g/l), methanol (50 g/l), phenol (2 g/l) and formaldehyde (7.5 g/l) proved that the isolate was an extremophilic organism. With high-density cultures, formaldehyde was totally removed at initial concentrations up to 7.5 g/l within 24 h, which is the highest biodegradation capability ever reported. Partial biodegradation of methanol (13 g/l) and diesel fuel (20 g/l) was also observed. Enzymatic studies revealed atypical methylotrophic pathway reactions, lacking alcohol oxidase, as compared with the conventional methylotroph Hansenula polymorpha. However, the activities of glutathione-dependent formaldehyde dehydrogenase, formaldehyde reductase, formate dehydrogenase and unspecific aldehyde dehydrogenase(s) were present. An additional glutathione-dependent aldehyde dehydrogenase activity was also detected. Metabolic and biochemical characteristics of the isolated yeast open up new possibilities for environmental biotechnology. Some potential applications in soil bioremediation and wastewater decontamination are discussed

Synthesis and characterization of ruthenium complexes with potential biological activity

Podczas reakcji dimerycznego związku rutenu(II) {[RuCl(η6-p-cymen]2(μ-Cl2)2} (DRu) z fosfinowymi pochodymi ciprofloksacyny (PCp), sparfloksacyny (PSf), lomefloksacyny (PLm) i norfloksacyny (PNr) w stosunku molowym 1:2, otrzymano nowe związki kompleksowe rutenu(II) o geometrii typu piano-stool. Struktury zsyntetyzowanych związków rozwiązano, wykorzystując metody rentgenowskiej analizy strukturalnej. Kompleksy zawierają w swojej budowie jon Ru2+, do którego koordynują pierścień p-cymenu, dwa jony Cl– i jedna cząsteczka fosfiny. Kompleksy scharakteryzowano, wykorzystując następujące techniki spektroskopowe – 1H NMR, 31P{1H} NMR oraz ESI-MS, czystość potwierdzono metodą analizy elementarnej. Potencjały redoks, równe średnio 1,08 V i 1,38 V w DMF (względem SEW) wskazują na możliwość uczestnictwa badanych kompleksów w reakcjach utleniania-redukcji w układach biologicznych. Fosfiny wykazują większe powinowactwo do ludzkiej albuminy i apo-transferyny niż ich kompleksy rutenu(II), przy czym zarówno ligandy fosfinowe jak i kompleksy nie wywołują zmian strukturalnych obu białek. Wykazano, że nowe kompleksy Ru(II) i PNr interkalują pomiędzy pary zasad podwójnej helisy DNA. Struktura DNA zostaje zmieniona w obecności fosfin, z kolei kompleksy indukują niewielkie jej zmiany. Hydrofobowe RuPCp i RuPNr zenkapsulowano w micelach (Pluronic P-123) z wydajnością enkapsulacji ok. 95%.Reaction of binuclear ruthenium(II) complex {[RuCl(η6-pcymen)]2(μ-Cl2)2} (DRu) with aminomethylphosphanes derived from ciprofloxacin (PCp), sparfloxacin (PSf), lomefloxacin (PLm) and norfloxacin (PNr) at molar ratio 1:2, results in formation of four new half-sandwich compounds. X-ray single crystal diffraction revealed that new ruthenium complexes consist of p-cymene ring, two Cl– ligands and one of phosphanes, all directly coordinated to Ru2+ centre. All complexes were characterized by selected spectroscopic methods – 1H NMR, 31P{1H} NMR and ESI-MS. Their purity was confirmed by elemental analysis. Determined redox potentials in DMF, equal to 1.08 V and 1.38 V (average for all complexes, vs. NHE), suggest their possible redox reactions in biological media. Phosphanes interact with human serum albumin and apo-transferrin more strongly than ruthenium complexes and does not change secondary structure of both proteins. Complexes and PNr demonstrate ability to intercalate between DNA bases, the rest of phosphanes do not intercalate at all. Furthermore, investigated phosphanes induce much stronger structure changes of DNA double helix than synthetized ruthenium complexes. One of Ru(II) compounds RuPCp was successfully encapsulated into Pluronic P-123 micelles, with encapsulation efficiency ca. 95%

Synthesis and characterization of new ruthenium(II) complexes containing morpholino aminomethylphosphanes

Dodatek dwukrotnego nadmiaru aminometylofosfiny P(CH2N(CH2CH2)2O)3 (A) lub PPh2(CH2N(CH2CH2)2O) (B) do roztworu binuklearnego związku rutenu {[RuCl(η6-p-cymen)]2(μ-Cl)2} (1) prowadzi do powstania nowych związków kompleksowych rutenu(II) typu „piano-stool”: [RuCl2(P(CH2N(CH2CH2)2O)3)(η6-p-cymen)] (2A) oraz [RuCl2(PPh2(CH2N(CH2CH2)2O))(η6-p-cymen)] (2B). W związkach tych, charakteryzujących się pseudo-oktaedryczną geometrią, centralny jon rutenu(II) jest skoordynowany z aromatycznym pierścieniem p-cymenu, dwoma anionami chlorkowymi oraz jedną cząsteczką aminometylofosfiny A lub B. Oba otrzymane związki scharakteryzowano za pomocą analizy elementarnej oraz metod spektroskopowych – magnetycznego rezonansu jądrowego (1H, 13C{1H}, 31P{1H}) i spektrometrii mas. Ponadto dla związku 2B określono strukturę krystaliczną na podstawie wyników otrzymanych metodą rentgenowskiej analizy strukturalnej.Addition of twofold excess of aminomethylphosphane P(CH2N(CH2CH2)2O)3 (A) or PPh2(CH2N(CH2CH2)2O) (B) into solution of binuclear ruthenium compound {[(η6-p-cymene)RuCl]2(μ-Cl)2} (1) leads to new ruthenium(II) piano-stool complexes: [RuCl2(P(CH2N(CH2CH2)2O)3)(η6-p-cymene)] (2A) and [RuCl2(PPh2(CH2N(CH2CH2)2O))(η6-p-cymene)] (2B). In these pseudo-octahedral compounds, central ruthenium(II) ion is coordinated with p-cymene aromatic ring, two chloride anions and one molecule of aminomethylphosphane A or B. Both obtained compounds were characterized by means of elemental analysis and spectroscopy methods – nuclear magnetic resonance (1H, 13C{1H}, 31P{1H}) and mass spectrometry. Moreover, 2B crystallographic structure was determined with single crystal X-ray diffraction

Polymeric micelle-mediated delivery of half-sandwich ruthenium(II) complexes with phosphanes derived from fluoroloquinolones for lung adenocarcinoma treatment

Novel half-sandwich ruthenium(II) complexes with aminomethyl(diphenyl)phosphine derived from fluoroloquinolones (RuPCp, RuPSf, RuPLm, RuPNr) were being investigated as alternatives to well-established metal-based chemotherapeutics. All compounds were characterized by elemental analysis, selected spectroscopic methods (i.e., absorption and fluorescence spectroscopies, ESI-MS, NMR, circular dichroizm), X-ray diffractometry, ICP-MS, and electrochemical techniques. To overcome low solubility, serious side effects connected with systemic cytotoxicity of ruthenium complexes, and acquiring the resistance of cancer cells, polymeric nanoformulations based on Pluronic P-123 micelles loaded with selected Ru(II) complexes were prepared and characterized. Resulting micelles (RuPCp_M, RuPNr_M) enabled efficient drug accumulation inside human lung adenocarcinoma (A549 tumor cell line), proved by confocal microscopy and ICP-MS analysis, allowing cytotoxic action. Studied complexes exhibited promising cytotoxicity in vitro with IC50 values significantly lower than the reference drug – cisplatin. The fluorescence spectroscopic data (CT-DNA titration, in vitro cell staining) together with analysis of DNA fragmentation (pBR322 plasmid, comet assay) provided clear evidence for the interaction with DNA inducing apoptotic cell death