Inclusion complexes of 3-(3-(2-chlorophenyl)prop-2-enoyl)-4-hydroxycoumarin with 2-hydroxypropyl-(-cyclodextrin: solubility and antimicrobial activity

The aim of the present study is to improve the solubility and antimicrobial activity of 3-(3-(2-chlorophenyl)prop-2-enoyl)-4-hydroxycoumarin by formulating its inclusion complexes with 2-hydroxypropyl-ß-cyclodextrin in solution and in solid state. The phase solubility study was used to investigate the interactions between 3-(3-(2-chlorophenyl)prop-2-enoyl)-4-hydroxycoumarin and 2-hydroxypropyl-ß-cyclodextrin and to estimate the molar ratio between them. The structural characterization of binary systems (prepared by physical mixing, kneading and solvent evaporation methods) was analysed using the FTIR-ATM spectroscopy. The antimicrobial activity of 3-(3-(2-chlorophenyl)prop-2-enoyl)-4-hydroxycoumarin and inclusion complexes prepared by solvent evaporation method was tested by the diffusion and dilution methods on various strains of microorganisms. The results of phase solubility studies showed that 3-(3-(2-chlorophenyl)prop-2-enoyl)-4-hydroxycoumarin formed the inclusion complexes with 2-hydroxypropyl-ß-cyclodextrin of AP type. The solubility of 3-(3-(2-chlorophenyl)prop-2-enoyl)-4-hydroxycoumarin was increased 64.05-fold with 50% w/w of 2-hydroxypropyl-ß-cyclodextrin at 37 oC. The inclusion complexes in solid state, prepared by the solvent evaporation method, showed higher solubility in purified water and in phosphate buffer solutions in comparison with 3-(3-(2-chlorophenyl)prop-2-enoyl)-4-hydroxycoumarin alone. The inclusion complexes prepared by solvent evaporation method showed higher activity on Bacillus subtilis and Staphylococcus aureus compared to uncomplexed 3-(3-(2-chlorophenyl)prop-2-enoyl)-4-hydroxycoumarin due to improved aqueous solubility, thus increasing the amount of available 3-(3-(2-chlorophenyl)prop-2-enoyl)-4-hydroxycoumarin that crosses the bacterial membrane.

Rapid identification of microbial contaminants in pharmaceutical products using a PCA/LDA-based FTIR-ATR method

Microbiological quality of pharmaceuticals is fundamental in ensuring efficacy and safety of medicines. Conventional methods for microbial identification in non-sterile drugs are widely used; however they can be time-consuming and laborious. The aim of this paper was to develop a chemometric-based rapid microbiological method (RMM) for identifying contaminants in pharmaceutical products using Fourier transform infrared with attenuated total reflectance spectrometry (FTIR-ATR). Principal components analysis (PCA) and linear discriminant analysis (LDA) were used to obtain a predictive model capable of distinguishing Bacillus subtilis (ATCC 6633), Candida albicans (ATCC 10231), Enterococcus faecium (ATCC 8459), Escherichia coli (ATCC 8739), Micrococcus luteus (ATCC 10240), Pseudomonas aeruginosa (ATCC 9027), Salmonella typhimurium (ATCC 14028), Staphylococcus aureus (ATCC 6538), and Staphylococcus epidermidis (ATCC 12228) microbial growth. FTIR-ATR spectra provide data on proteins, DNA/RNA, lipids, and carbohydrates constitution of microbial growth. Microbial identification provided by PCA/LDA based on FTIR-ATR method were compatible with those obtained using traditional microbiological methods. The chemometric-based FTIR-ATR method for rapid identification of microbial contaminants in pharmaceutical products was validated by assessing the sensitivity (93.5%), specificity (83.3%), and limit of detection (17-23 CFU/mL of sample). Therefore, we propose that FTIR-ATR spectroscopy may be used for rapid identification of microbial contaminants in pharmaceutical products and taking into account the samples studied

Biological activity of synthesized 5-{1-[(4-chlorophenyl)sulfonyl]piperidin-4-yl}-2-mercapto-1, 3, 4-oxadiazole derivatives demonstrated by in silico and BSA binding studies

We synthesized a series of compounds bearing pharmacologically important 1,3,4-oxadiazole and piperidine moieties. Spectral data analysis by 1H-NMR, 13C-NMR, IR and EI-MS was used to elucidate the structures of the synthesized molecules. Docking studies explained the different types of interaction of the compounds with amino acids, while bovine serum albumin (BSA) binding interactions showed their pharmacological effectiveness. Antibacterial screening of these compounds demonstrated moderate to strong activity against Salmonella typhi and Bacillus subtilis but only weak to moderate activity against the other three bacterial strains tested. Seven compounds were the most active members as acetyl cholinesterase inhibitors. All the compounds presented displayed strong inhibitory activity against urease. Compounds 7l, 7m, 7n, 7o, 7p, 7r, 7u, 7v, 7x and 7v were highly active, with respective IC50 values of 2.14±0.003, 0.63±0.001, 2.17±0.006, 1.13±0.003, 1.21±0.005, 6.28±0.003, 2.39±0.005, 2.15±0.002, 2.26±0.003 and 2.14±0.002 µM, compared to thiourea, used as the reference standard (IC50 = 21.25±0.15 µM). These new urease inhibitors could replace existing drugs after their evaluation in comprehensive in vivo studies.

Lipopeptide production by Bacillus subtilis R1 and its possible applications

Abstract The possible application of a bacterial strain - Bacillus subtilis R1, isolated from an oil contaminated desert site in India, as biocontrol agent and its biosurfactant in microbial enhanced oil recovery are discussed. The biosurfactant production in minimal medium was carried out at different temperatures and salt concentrations, where it produced an efficient biosurfactant at 30-45 °C and in presence of up to 7% salt. It significantly reduced the surface tension from 66 ± 1.25 mN/m to 29 ± 0.85 mN/m within 24 h. In order to enhance the biosurfactant production, random mutagenesis of B. subtilis R1 was performed using chemical mutagen - ethyl methanesulfonate. Majority of the isolated 42 mutants showed biosurfactant production, but the difference was statistically insignificant as compared with parent strain R1. Therefore none of the mutants were selected for further study, and only parent strain R1 was studied. The biosurfactant was quite stable under harsh conditions for up to 10 days. The biosurfactant was extracted and characterized as similar to the lipopeptide group - surfactins and fengycin. The crude oil displacement experiments using biosurfactant broth in sand pack glass columns showed 33 ± 1.25% additional oil recovery. The strain also showed inhibition of various plant pathogenic fungi on potato dextrose agar medium.

Biological control of the grapevine diseases ‘grey mold’ and ‘powdery mildew’ by Bacillus B27 and B29 strains.

Uncinula necator and Botrytis cinerea are the most destructive pathogens of the grapevine in Tunisia and elsewhere. We used two strains of Bacillus subtilis group, B27 and B29 to control powdery mildew and the grey mold disease of the grapevine. Green house experiments showed that B29 and B27 strains of the bacteria efficiently reduced the severity of powdery mildew up to 50% and 60%, respectively. Further, they decreased Botrytis cinerea development on grape leaf by 77% and 99%, respectively. The mode of action has been shown to be chitinolytic. These two bacteria showed significant production of total proteins discharged into the culture medium. Determination of some chitinolytic enzymes revealed the involvement of N-acetyl glucosaminidase (Nagase), the chitin-1,4-chitobiosidase (Biase) and endochitinase in degrading the mycelium of B. cinerea.

Potentialities of newly isolated Bacillus subtilis and Lactobacillus sp for curd preparation and a comparative study of its physico-chemical parameters with other marketed curds.

Two Bacillus sp. were isolated from the local fermented milk and identified on the basis 16S rRNA sequence profile as Bacillus subtilis AKL1 and by biochemical process as Lactobacillus acidophilus AKL2. These isolates were used as fresh inoculums for curd preparation individually and in combinations. Different physico-chemical and therapeutic properties of the newly prepared curd were examined and compared with marketed local (sweet and sour) and branded (Mother Dairy and Thackar) curds. The total hydrolyzed peptides, free amino acids, lactic acid were significantly higher, whereas, total solid, ash content, syneresis and free reducing sugar were lower in the curd prepared by a mixture of AKL1 and AKL2 (0.5:0.5, v/v). The antioxidant activity against ABTS+, DPPH•, OH• and Fe3+ were also higher in the newly formulated curd. Polyphenols (85.5µg/g), flavonoids (12.5µg/g) and free aromatic amino acids contents were also higher in AKL1+AKL2. All these components prevent excess protein oxidation that was revealed by SDS-PAGE. The curd also exhibited potent antimicrobial activity against some entero-pathogens like Clostridium perfringens, Escherichia coli, Shigella dysentery, Vibrio cholerae and Staphylococcus aureus. It can be concluded that the combination of these Lactobacillus sp. will be a fruitful inoculum for the preparation of curd having better health promoting effects.

Evaluación de los cambios en algunas propiedades físicas y químicas de un ultisolpor efecto del bacillus subtilis / Evaluation of the changes in the some physical and chemical properties of an ultisol for effect of the bacillus subtilis

El interés en estudios de biorremediación de suelos deteriorados por sobreexplotación y uso indiscriminado de agroquímicos se debe a que alteran la microflora, el sistema de autorregulación y la sustentabilidad en el largo plazo. Un Ultisol, suelo de baja fertilidad química y sometido a reducción de tamaño de agregados (<2,0 mm), solarizado para reducir la población microbiológica, fue inoculado con la bacteria Basillus subtilis en concentraciones de 106, 107, 108, 109 unidades formadoras de colonias (ufc). En 120 días se observó un incremento importante en magnitud de: estabilidad estructural, tamaño promedio de agregados, pH, fósforo disponible y una disminución del aluminio intercambiable. Estas variaciones en la respuesta estuvieron relacionadas con la actividad de los microorganismos en el suelo, y responden a la capacidad de solubilización de minerales por la bacteria, de producción de condiciones alcalinas y de biofilms, que unidos al aumento de biomasa de raíces de la planta, mucílagos y carbohidratos, coadyuvan en la formación de agregados estables y de mayor tamaño. Las propiedades físicas y químicas al final del experimento se estabilizaron en valores mayores a los encontrados en el suelo inicial, produciendo un efecto positivo general sobre el mismo, desde el punto de vista de la fertilidad global, al aumentar el fósforo disponible, disminuir la acidez intercambiable e incrementar la estabilidad y el tamaño promedio de agregados del suelo a corto plazo.

The interest about the studies of bioremediation in deteriorated soils under and over exploitation, indiscriminate use of agrochemicals that alter the microflora, the self regulation system and the sustainability in the long term. An Ultisol, with poor chemical properties and low fertility, because the reduction of aggregates size (< 2.0 mm). Solarize to reduce the microbiological population, it was inoculated with the bacteria Basillus subtilis in concentrations of 106, 107, 108, 109 colony forming units (cfu). In 120 days important increment in magnitude of: structural stability, size average of aggregates, pH, available phosphorus. Also diminish the interchangeable aluminum. These variations in the answer were related with the activity of the microorganisms in the soil, and it responds so that solubilisation capacity of minerals because the bacteria activity, production of alkaline conditions and biofilms too. Furthermore the increase of plant roots biomass, mucilages and carbohydrates cooperate in the formation of stable aggregates and bigger size of them. The physical and chemical properties at the end of the experiment were stabilized in bigger values than found in the initial soil. Confirming a positive general effect overall the soil, since the point of view of the global fertility, when increasing the available phosphorus, reduce the interchangeable acidity and increase the soil stability and average size aggregates in the short term.