Synthesis,micellisation and interaction of novel quaternary ammonium compounds derived from l-Phenylalanine with 1,2-dipalmitoyl-sn-glycero-3-phosphocholine as model membrane in relation to their antibacterial activity,and their selectivity over human red blood cells

A series of quaternary ammonium compounds (QUATS) derived from l-Phenylalanine have been synthesized and their antibacterial efficiencies were determined against various strains of Gram-positive and Gram-negative bacteria. The antibacterial activity increased with increasing chain length, exhibiting a cut-off effect at C₁₄ for Gram-positive and C₁₂ for Gram-negative bacteria. The l-Phenylalanine QUATS displayed enhanced antibacterial properties with a higher cut-off point compared to their corresponding l-Phenylalanine ester hydrochlorides. The CMC was correlated with the MIC, inferring that micellar activity contributes to the cut-off effect in antibacterial activity. The hemolytic activities (HC₅₀) of the QUATS against human red blood cells were also determined to illustrate the selectivity of these QUATS for bacterial over mammalian cells. In general, the MIC was lower than the HC₅₀, and assessment of the micellar contribution to the antibacterial and hemolytic evaluation in TBS as a common medium confirmed that these QUATS can act as antibacterial, yet non-toxic molecules at their monomer concentrations. The interaction of the QUATS with the phospholipid vesicles (1,2-dipalmitoyl-sn-glycero-3-phosphocholine, DPPC) in the presence of 1-anilino-8-naphthalene sulfonate (ANS) and 1,6-diphenyl-1,3,5-hexatriene (DPH) as fluorescence probes showed that the presence of the quaternary ammonium moiety causes an increase in hydrophobic interactions, thus causing an increase in antibacterial activity.     

Effect of Tween 80 on the production of curdlan by Alcaligenes faecalis ATCC 31749

This study aims to investigate the effects of Tween 80 on curdlan production, cell growth, and glucosyltransferase activity. The addition of Tween 80 to the culture medium increased curdlan production. However, curdlan production did not increase further when excessive Tween 80 (>0.3% Tween 80) was added to the culture medium. The addition of Tween 80 to the culture medium did not affect cell growth. The glucosyltransferase activity involved in the curdlan synthesis increased with the increase of Tween 80 concentration. The glucosyltransferase activity did not increase further when excessive Tween 80 (>0.3% Tween 80) was added to the culture medium. Maximum curdlan was observed at day 5 and then levelled off. The biomass continued to increase until the end of the experimental period (6 d). Maximum glucosyltransferase activity was also observed at day 5 and decreased thereafter. The results indicate that the enhanced curdlan production by Tween 80 is highly correlated with glucosyltransferase activity.     

Isolation and structural characterization of biosurfactant produced by an alkaliphilic bacterium Cronobacter sakazakii isolated from oil contaminated wastewater

Production of biosurfactant from an alkaliphilic bacterium Cronobacter sakazakii (accession no. JN398668) was screened by haemolytic assay, emulsifying activity and surface tension measurement. Biosurfactant, comprised of total sugars (73.3%), reducing sugars (1.464%), protein (11.9%), uronic acid (15.98%) and sulfate (6.015%), showed low viscosity with pseudoplastic rheological behavior and exhibited significant emulsification activity with oils and hydrocarbons. A series of low and mid range mass peaks (m/z) corresponding to mono-, di-, tri- and oligosaccharides were detected in the positive ion reflector mode of MALDI TOF-TOF MS. GC-MS analysis revealed composition of monosaccharide moieties (w/w) viz. glucose (14%), mannose (24%), galactose (14%), xylose (20%) and arabinose (1.9%). ¹H NMR, FT-IR and EDX analyses confirmed the characteristic various functional groups, bonds and elements respectively. Thermostability (up to 260 °C) and CI (0.456) were determined by TG and DSC analyses. Inherent properties of biosurfactant make it a potential candidate for bioremediation of oil and hydrocarbons.     

Serratia sp. SVGG16: a promising biosurfactant producer isolated from tropical soil during growth with ethanol-blended gasoline

The strain SVGG16 isolated from tropical soil in Brazil and identified as Serratia sp. was selected as a promising biosurfactant producer using atypic growth conditions including ethanol-blended gasoline as the sole carbon source. Important parameters for biosurfactant production were selected through experimental design. Results demonstrated that this strain was able to reduce surface tension of the medium to 34 mN m⁻¹, showing potential to be used in bioremediation processes.     

Bacicyclin,a new antibacterial cyclic hexapeptide from Bacillus sp. strain BC028 isolated from Mytilus edulis

A new cyclic hexapeptide, cyclo-(Gly-Leu-Val-IIe-Ala-Phe), named bacicyclin (1), was isolated from a marine Bacillus sp. strain associated with Mytilus edulis. The sequences of the amino acid building blocks of the cyclic peptide and its structure were determined by 1D- and 2D-NMR techniques. Marfey’s analysis showed that the amino acid building blocks had L-configuration in all cases except for alanine and phenylalanine, which had D-configuration. Bacicyclin (1) exhibited antibacterial activity against the clinically relevant strains Enterococcus faecalis and Staphylococcus aureus with minimal inhibitory concentration values of 8 and 12?µM, respectively. These results demonstrate the potential of marine bacteria as a promising source for the discovery of new antibiotics.     

Homoagmatine from Lathyrus sativus seedlings

A new guanidino amine has been isolated from Lathyrus sativus seedlings and chararterized as homoagmatine on the basis of various physico-chemical criteria including IR spectrum and comparison with that chemically synthesized. Homoagmatine is accumulated in the embryos axis while its precursor, homoarginine, is lost from the cotyledons. However, there was a progressive increase in homoarginine content of the embryo axis during development. Since the amine content of the whole seedlings corresponded to nearly 20–25 % of net decrease in homoarginine levels, it is concluded that the catabolism of homoarginine through homoagmatine represents a major pathway of metabolism of the arnino acid.     

Microbial biosurfactant mediated removal and/or solubilization of crude oil contamination from soil and aqueous phase: An approach with Bacillus licheniformis MTCC 5514

This study highlights the role of marine microbial biosurfactants on solubilization/removal of crude-oil contamination from four different soils in an aqueous phase. Soil of four different types, viz., sandy, fine sand soil, clay, and clay loam, were collected and saturated with crude oil. Marine isolate MTCC 5514 (Bacillus licheniformis) was chosen for the study and comparisons were made with synthetic surfactants and commercially available biosurfactant. In-situ studies were carried out with different percentages of crude oil to assess the growth and the percentage removal of oil. For ex-situ studies, soils were pre-saturated with crude oil and then treated with the chosen biosurfactant at a 10% concentration level using flask and column methods. After time intervals of 30–120 min, samples were collected and then subjected to extraction with hexane and the percentage removal was calculated. Results revealed, at 2% concentration of crude oil, that complete solubilization was achieved. With regard to ex-situ studies, clay soil absorbed the maximum percentage of oil compared to other soil types, and with regard to the removal, all the synthetic surfactants showed <60% removal irrespective of soil type. In the case of biosurfactants even at 10% concentration, >85% removal was achieved.     

The mucus of Sabella spallanzanii (Annelida, Polychaeta): Its involvement in chemical defence and fertilization success

In several species mucus typically forms a slippery coating that performs many specialised roles including the defence against infection and protection for eggs. In the present study we focused on the mucus of the polychaete Sabella spallanzanii in order to ascertain its involvement in chemical defence and fertilization success. The observations were carried out over an annual cycle according to different phases of the reproductive cycle. Mucus of S. spallanzanii showed a natural lysozyme-like activity with a maximum at pH 6.0 and I = 0.175. This activity was present in all the sampling times except for January (T₄) which corresponds to the reproductive period of this species. The electrophoretic analysis revealed at least ten major protein bands, with molecular weights ranging from 16 to 90 kDa, and six minor components, with molecular weights ranging from 14 to 116 kDa. This pattern was reported all over the year except for the reproductive period when not only the lysozyme-like activity decreased but the number of protein bands was also reduced. During this period microscopical observations of the mucus indicated that only fertilised eggs enveloped in the mucus were released out of the tubes where the worms live. This supports previous hypothesis that fertilization does not occur in the water column but it takes place in the tube just after egg release (in situ fertilization). This fertilization strategy, not involving egg dispersal in the water column, ensures a high success of fertilization. The mucus of S. spallanzanii therefore represents the ideal mean where eggs are released and fertilized as well as a compartment involved in defending the worms from bacterial attack serving as medium into which the antibacterial substances are exuded. The defensive role of lysozyme is important taking into account that this polychaete lives in eutrophic environments such as harbours where bacteria, including pathogens to man and marine organism, are abundant.     

金荞麦的抑菌活性研究

采用试管稀释法和管碟法分别研究了金荞麦(Fagopyrum dibotrys )根状茎和茎叶不同溶剂提取物对6种细菌和13种真菌的体外抑菌作用。结果显示,金荞麦对细菌和真菌均有一定的抑制作用。对细菌的抑菌效果表明:各种提取物对金黄色葡萄球菌、大肠杆菌、枯草芽孢杆菌、苏云芽孢杆菌、卡拉双球菌都有明显的抑菌作用,但是对5406放线菌只有根乙醇提取物和茎叶水提取物有一定的抑制效果。对真菌的抑菌效果表明:各种提取物对鞭毛菌、白色念珠菌、松赤枯病菌、玉米纹枯病菌、油菜菌核病菌、玉米弯胞杆菌、小麦赤霉病菌、绿色木霉都有明显的抑菌作用。     

Bioactive polyhydroxylated sterols from the marine sponge Haliclona crassiloba

Four new polyhydroxylated sterols, named halicrasterols A–D (1–4), together with six known analogs (5–10) were isolated from the marine sponge Haliclona crassiloba. Compounds 1 and 2 represented rare examples of steroids featuring 17(20)E-double bonds. The structures of 1–10 were elucidated by spectroscopic analysis and comparison with reported data. This is the first report of a steroid profile for this species. The antimicrobial activities of 1–10 were evaluated against a panel of bacterial and fungal strains in vitro, and compounds 4 and 9 showed moderate activity against some of the Gram-positive strains with MICs ranging from 4 to 32 μg/mL.     

海南产木薯茎化学成分研究

从木薯(Manihot esculenta Crantz)茎的乙醇提取物中分离得到6个化合物,通过波谱分析,分别鉴定为呋喃(1)、肥牛木素(2)、3-吲哚甲酸(3)、3,9,13-megastigmanetriol (4)、穗花杉双黄酮(5)、yucalexin P-21 (6),其中化合物1~5为首次从该植物中分离得到。用滤纸片琼脂扩散法测定了这些化合物的抗菌活性,结果表明化合物1、3、4和6对耐甲氧西林金黄色葡萄球菌(MRSA)和金黄色葡萄球菌均有抑制作用。     

A colorimetric determination for glycosidic and bile salt-based detergents: applications in membrane protein research

Detergents are crucial to the isolation of integral membrane proteins. During membrane protein purification, it is useful to accurately quantify detergent, especially if concentration steps have been used. Previously, this has been difficult and time-consuming. We present a simple, rapid, and sensitive procedure for the quantification of glycosidic and bile salt-based detergents such as dodecylmaltoside, octylglucoside, and CHAPS. The method directly quantifies sugar or cholate moieties via colorimetric reactions with phenol and sulfuric acid. A number of detergents have been screened, and the assay has been validated in the presence of commonly used reagents. In addition to determining the overall detergent concentration in solution, the procedure allows accurate quantification of specific binding of glycosidic or bile salt-based detergents to purified membrane proteins. Both the colorimetric method and the radiometric 14C method were used to determine detergent binding to two integral membrane proteins: the cytochrome cbb3 oxidase from Pseudomonas stutzeri and the turkey beta-adrenergic receptor. Both methods gave similar results. After separating monomeric glycosidic detergent from micellar solutions by ultrafiltration, we used the colorimetric method to determine the concentration of monomeric detergent present. We observed that values obtained are in close agreement with previously determined critical micelle concentrations.     

Antibacterial and antioxidant activities of four kaempferol methyl ethers isolated from Dodonaea viscosa Jacq. var. angustifolia leaf extracts

Fractionation of dichloromethane and acetone fractions obtained by serial extraction from the leaf powder of Dodonaea viscosa Jacq. var. angustifolia (Sapindaceae) resulted in the isolation of four kaempferol methyl ethers. The compounds were identified by spectral data (¹H NMR, ¹³C NMR and MS) as: 3, 5, 7-trihydroxy-4'-methoxyflavone (1); 5, 7, 4'-trihydroxy-3, 6-dimethoxyflavone (2); 5, 7-dihydroxy-3, 6, 4'-trimethoxyflavone (santin) (3); and 5-hydroxy -3, 7, 4'-trimethoxyflavone (4) together with 3,4',5,7-tetrahydroxy flavone (kaempferol) (5). Antioxidant potential of the compounds was evaluated using a DPPH spectrophotometric assay, while antibacterial activity was determined using a serial dilution microplate technique. The isolates demonstrated varying degrees of antioxidant and antibacterial activities. Of all the compounds investigated, compounds 1 and 5 demonstrated some antioxidant activity (EC₅₀ = 75.49 ± 1.76 µM and 35.06 ± 0.85 respectively) but lower than l-ascorbic acid (EC₅₀ = 13.55 ± 0.28 µM) used as a standard antioxidant agent. The minimum inhibitory concentration (MIC) of isolated compounds against Staphylococcus aureus, Enterococcus faecalis, Escherichia coli and Pseudomonas aeruginosa varied from 16 µg/ml to more than 250 µg/ml. Some structure activity relationships could be established for these compounds.     

Pyrone derivatives from the marine-derived fungus Nigrospora sp. PSU-F18

Pyrones, named nigrosporapyrones A-D (1-4), and five known compounds were isolated from the marine-derived fungus Nigrospora PSU-F18. Their structures were elucidated on the basis of spectroscopic evidence. The antibacterial activity against the standard Staphylococcus aureus ATCC 25923 and methicillin-resistant S. aureus was evaluated.     

Purification,characterization and immunostimulatory activity of polysaccharide from Cipangopaludina chinensis

In this study, we investigated the purification, preliminary characterization and immunostimulatory activity in vivo of polysaccharide from Cipangopaludina chinensis (CCPS). Firstly, crude CCPS was prepared by hot water extraction. And the crude CCPS was sequentially purified by chromatography of DEAE-52 and Sephadex G-100, resulting in two purified fractions of CCPS-1 and CCPS-2. We found the two fractions were homogeneous heteropolysaccharides mainly composed of rhamnose and glucose with the average molecular weight of 226 and 235 kDa, respectively. CCPS-2 was quite different from CCPS-1. It had much higher content of uronic acid and sulfuric radical. For immunostimulatory activity in vivo, crude CCPS could significantly increase the thymus and spleen indices, enhance the macrophage function, and increase the level of serum hemolysin in cyclophosphamide-treated mice, suggesting CCPS had a potent immunostimulatory activity and could be explored as a potential natural immunomodulatory agent     

South African Lippia herbal infusions: Total phenolic content,antioxidant and antibacterial activities

Lippia javanica and Lippia scaberrima are used as herbal remedies and are commercially traded as health teas in southern Africa under the brands “Mosukujane” and “Musukudu”, respectively. This study evaluates the relationship between the presence of phenolic compounds and the antioxidant activities of infusions prepared from four Lippia species (L. javanica, L. scaberrima, L. rehmannii and L. wilmsii) indigenous to South Africa. The antioxidant activities of the infusions, determined by the 2,2-diphenylpycrylhydrazyl (DPPH) method, were also compared to those of popular black, green and herbal tea brands. Of the four indigenous species, infusions of L. javanica and L. wilmsii exhibited the highest antioxidant activities (EC₅₀: 358 and 525 µg/ml, respectively) and contained the most phenolic compounds (14.8 and 14.5 mg/ml of dry weight gallic acid equivalent, respectively). Antibacterial activities of methanolic extracts of the four Lippia species were determined against four human pathogens (Staphylococcus aureus, Enterococcus faecalis, Escherichia coli and Pseudomonas aeruginosa). The extract of L. javanica was the most active against all the pathogens tested. Those Lippia species (L. javanica and L. wilmsii) previously reported to produce higher levels of the pharmacologically active phenylethanoid glycosides verbascoside and isoverbascoside, portrayed stronger antioxidant and antibacterial activities. This study gives credence to the use of infusions of these Lippia species for their general health benefits.     

Glycosylated liposomes against Helicobacter pylori: Behavior in acidic conditions

Helicobacter pylori was isolated in 1982 and confirmed as a gastric pathogenic agent at the end of the 1980s. The present work deals with liposomes formulations in which are incorporated cholesteryl tetraethylene glycol oside as model ligands for H. pylori adhesins. This study is devoted to the behavior of liposomes in gastric conditions. The glycosylated vesicles are stable and the pH of the internal aqueous compartment remains close to 4 even through more acidic conditions are imposed to the external phase (pH 1.2-2). Such a pH gradient depends essentially on the nature of phospholipids used and is not extensively affected by the incorporation of the targeting agent. These aspects are particularly important to the development of liposome formulations against H. pylori, bacteria sensitive to antibiotics which are unstable in very acidic conditions.     

金荞麦的抑菌活性研究

采用试管稀释法和管碟法分别研究了金荞麦(Fagopyrum dibotrys)根状茎和茎叶不同溶剂提取物对6种细菌和13种真菌的体外抑菌作用。结果显示,金荞麦对细菌和真菌均有一定的抑制作用。对细菌的抑菌效果表明:各种提取物对金黄色葡萄球菌、大肠杆菌、枯草芽孢杆菌、苏云芽孢杆菌、卡拉双球菌都有明显的抑菌作用,但是对5406放线菌只有根乙醇提取物和茎叶水提取物有一定的抑制效果。对真菌的抑菌效果表明:各种提取物对鞭毛菌、白色念珠菌、松赤枯病菌、玉米纹枯病菌、油菜菌核病菌、玉米弯胞杆菌、小麦赤霉病菌、绿色木霉都有明显的抑菌作用。