Regioselective preparation of 5-hydroxypropranolol and 4′-hydroxydiclofenac with a fungal peroxygenase

An extracellular peroxygenase of Agrocybe aegerita catalyzed the H₂O₂-dependent hydroxylation of the multi-function beta-adrenergic blocker propranolol (1-naphthalen-1-yloxy-3-(propan-2-ylamino)propan-2-ol) and the non-steroidal anti-inflammatory drug diclofenac (2-[2-[(2,6-dichlorophenyl)amino]phenyl]acetic acid) to give the human drug metabolites 5-hydroxypropranolol (5-OHP) and 4′-hydroxydiclofenac (4′-OHD). The reactions proceeded regioselectively with high isomeric purity and gave the desired 5-OHP and 4′-OHD in yields up to 20% and 65%, respectively. ¹⁸O-labeling experiments showed that the phenolic hydroxyl groups in 5-OHP and 4′-OHD originated from H₂O₂, which establishes that the reaction is mechanistically a peroxygenation. Our results raise the possibility that fungal peroxygenases may be useful for versatile, cost-effective, and scalable syntheses of drug metabolites.     

Exploiting the natural poly(3-hydroxyalkanoates) production capacity of Antarctic Pseudomonas strains: from unique phenotypes to novel biopolymers

Journal of Industrial Microbiology & Biotechnology - Extreme environments are a unique source of microorganisms encoding metabolic capacities that remain largely unexplored. In this work, we...     

Design,Synthesis and In Vitro Activity of Anticancer Styrylquinolines. The p53 Independent Mechanism of Action

A group of styrylquinolines were synthesized and tested for their anti-proliferative activity. Anti-proliferative activity was evaluated against the human colon carcinoma cell lines that had a normal expression of the p53 protein (HCT116 p53^+/+) and mutants with a disabled TP53 gene (HCT116 p53^-/-) and against the GM 07492 normal human fibroblast cell line. A SAR study revealed the importance of Cl and OH as substituents in the styryl moiety. Several of the compounds that were tested were found to have a marked anti-proliferative activity that was similar to or better than doxorubicin and were more active against the p53 null than the wild type cells. The cellular localization tests and caspase activity assays suggest a mechanism of action through the mitochondrial pathway of apoptosis in a p53-independent manner. The activity of the styrylquinoline compounds may be associated with their DNA intercalating ability.     

Synthesis of New Styrylquinoline Cellular Dyes,Fluorescent Properties,Cellular Localization and Cytotoxic Behavior

New styrylquinoline derivatives with their photophysical constants are described. The synthesis was achieved via Sonogashira coupling using the newly developed heterogeneous nano-Pd/Cu catalyst system, which provides an efficient synthesis of high purity products. The compounds were tested in preliminary fluorescent microscopy studies to in order to identify their preferable cellular localization, which appeared to be in the lipid cellular organelles. The spectroscopic properties of the compounds were measured and theoretical TD-DFT calculations were performed. A biological analysis of the quinolines that were tested consisted of cytotoxicity assays against normal human fibroblasts and colon adenocarcinoma cells. All of the compounds that were studied appeared to be safe and indifferent to cells in a high concentration range. The presented results suggest that the quinoline compounds that were investigated in this study may be valuable structures for development as fluorescent dyes that could have biological applications.     

Centipede (Chilopoda) richness and diversity in the Bug River valley (Eastern Poland)

The main aim of the survey was to describe the diversity and richness of Chilopoda in the selected area of the Bug River valley. The study sites were located in two regions differing in the shape of the valley, the presence of thermophilous habitats and the size of riparian forests. Pitfall traps were used as a sampling method. As a result, 444 specimens belonging to 12 centipede species of two orders – Geophilomorpha (four species) and Lithobiomorpha (eight species) were caught. Lithobius (Monotarsobius) curtipes C.L.Koch, 1847, Pachymerium ferrugineum (C.L.Koch, 1835), Lamyctes (Lamyctes) emarginatus (Newport, 1844) and Lithobius (Monotarsobius) dudichi Loksa, 1947 were the most common and the most numerous species. Of particular note is Lithobius dudichi found in Poland for the first time and previously known based on a single specimen. Two to 10 Chilopoda species were found in each habitat under investigation. The greatest species richness was found in thermophilous thickets (10 species), sandy grasslands (eight), xerothermic grasslands (eight) and mesic meadows (six). The fewest number of species (two) was found in rushes at oxbows and in wet meadows. We found differences in the species composition and the number of Chilopoda between the lower (102 specimens, six species) and the middle (324 specimens, 11 species) section of the river valley. Our results confirm the need to protect xerothermic habitats, unique almost throughout entire Central Europe, which due to their distribution and their small area covered are fairly easily subject to the process of destruction.     

Tetracycline-resistant bacteria as indicators of antimicrobial resistance in protected waters—The example of the Drwęca River Nature Reserve (Poland)

The objective of this study was to determine the suitability of Tet^R tetracycline-resistant bacteria as potential indicators of drug resistance, a parameter of the microbiological quality of river waters in natural reserves which are threatened by man-made pollution. The microbiological assays covered a 15-km long section of the upper reach of the Drw?ca River (Poland), a part of the European Ecological “Natura 2000” Network of nature protected areas. The quality of the investigated ecosystem was affected by surface runoffs from the river's agricultural catchment as well as outflows from three fish farms. The counts of Tet^R bacteria, incubated at 14 °C and 28 °C on TSA medium with sheep blood and tetracycline, were determined in river water samples. The highest counts of both bacterial groups were determined in samples collected from sites behind fish farms. A statistical analysis of the abundance of Tet^R14 °C and Tet^R28 °C bacteria revealed significant differences in the size of Tet^R28 °C populations at the studied sampling sites (p = 0.0011), which is why hemolytic bacteria of this group (HemTet^R28 °C) were selected for further investigations. The predominant strains in the group of 86 HemTet^R28 °C isolates obtained by 16S rRNA gene sequencing were Pseudomonas fluorescens (42 isolates) and Aeromonas hydrophila (28 isolates). Analyses of the identified HemTet^R28 °C strains demonstrated MIC ≥256 μg/ml in more than 50% isolates. The MAR index of HemTet^R28 °C was in the range of 0.67 at the control site to 1 at sites behind fish farms. The results suggest that tetracycline-resistant bacteria, in particular HemTet^R28 °C, are a reliable indicator of antimicrobial resistance and the microbial quality of surface waters polluted due to human activity. The above can be attributed to several factors: (I) the highest percentage share of HemTet^R28 °C among HPC28 °C was noted at sites behind fish farms, (II) tetracycline-resistant bacteria quickly respond to environmental changes, as demonstrated by the high level of resistance to tetracycline and a very high MAR index, and (III) genera/species that are easy to culture under laboratory conditions dominate in the qualitative and quantitative composition of the studied bacteria.     

Effect of medium composition,genotype and age of explant on the regeneration of hexaploid plants from endosperm culture of tetraploid kiwiberry (Actinidia arguta)

Plant Cell, Tissue and Organ Culture (PCTOC) - Endosperm, an ephemeral and storage tissue, serves as a source of nutrition and protection during embryo development and germination. It can be used...     

Transformation of Nasturtium officinale, Barbarea verna and Arabis caucasica for hairy roots and glucosinolate-myrosinase system production

Hairy roots of Nasturtium officinale, Barbarea verna and Arabis caucasica with active glucosinolate-myrosinase system were obtained after transformation with Agrobacterium rhizogenes. Hairy roots of N. officinale produced phenylalanine-derived gluconasturtiin and glucotropaeolin (max. 24 and 7 mg g⁻¹ DW). B. verna and A. caucasica hairy roots produced gluconasturtiin (max. 41 mg g⁻¹ DW) and methionine-derived glucoiberverin (max. 32 mg g⁻¹ DW), respectively. Treatment of the roots with amino acid precursors of glucosinolate or/and cysteine biosynthesis increased levels of glucosinolate production, combinations of phenylalanine with cysteine (for gluconasturtiin and glucotropaeolin) and methionine with o-acetylserine (for glucoiberverin) were the most effective.     

Nickel(II)-substituted azurin I from Alcaligenes xylosoxidans as characterized by resonance Raman spectroscopy at cryogenic temperature

Metal-substituted blue copper proteins (cupredoxins) have been successfully used to study the effect of metal-ion identity on their active-site properties, specifically the coordination geometry and metal–ligand bond strengths. In this work, low-temperature (77 K) resonance Raman (RR) spectra of the blue copper protein Alcaligenes xylosoxidans azurin I and its Ni(II) derivative are reported. A detailed analysis of all observed bands is presented and responsiveness to metal substitution is discussed in terms of structural and bonding changes. The native cupric site exhibits a RR spectrum characteristic of a primarily trigonal planar (type 1) coordination geometry, identified by the ν(Cu–S)_Cys markers at 373, 399, 409, and 430 cm⁻¹. Replacement of Cu(II) with Ni(II) results in optical and RR spectra that reveal (1) a large hypsochromic shift in the main (Cys)S → M(II) charge-transfer absorption from 622 to 440 nm, (2) greatly reduced metal–thiolate bonding interaction, indicated by substantially lower ν(Ni–S)_Cys stretching frequencies, (3) elevation of the cysteine ν(C _β –S) stretching, amide III, and ρ _s(C _β H₂) scissors vibrational modes, and (4) primarily four-coordinated, trigonally distorted tetrahedral geometry of the Ni(II) site that is marked by characteristic ν(Ni–S)_Cys stretching RR bands at 347, 364, and 391 cm⁻¹. Comparisons of the electronic and vibrational properties between A. xylosoxidans azurin I and its closely structurally related azurin from Pseudomonas aeruginosa are made and discussed. For cupric azurins, the intensity-weighted average M(II)–S(Cys) stretching frequencies are calculated to be ν(Cu–S)_iwa = 406.3 and 407.6 cm⁻¹, respectively. These values decreased to ν(Ni–S)_iwa = 359.3 and 365.5 cm⁻¹, respectively, after Ni(II) → Cu(II) exchange, suggesting that the metal–thiolate interactions are similar in the two native proteins but are much less alike in their Ni(II)-substituted forms.