Biochemical Characterization of Plasma Membrane Vesicles of Cyanophora paradoxa*

Plasma membrane vesicles (PMVs) were isolated from Cyanophora paradoxa Korshikov (Claucocystophyta) and purified by differential and sucrose density gradient centrifugation. PMVs were identified ultrastructurally and biochemically using marker enzymes. Analysis of monosaccharides revealed a sugar content of 37.3% of dry weight of delipidated PMVs with high amounts of fucose (28 mol% of total carbohydrate), galactose (23%), mannose (21%) and the minor constituents xylose, rhamnose, glucose and N-acetylglucosamine. Proteins were analyzed by SDS-PACE. More than 20 polypeptides were resolved and several were identified as glycopeptides, as shown by PAS staining and lectin blots. Fucose linked α-1→6 to N-acetylglucosamine (AAA-binding), mannose linked α-1→2,3,6 to mannose (CNA-binding) and terminally bound galactose (RCA-binding) were identified on several polypeptides. Sterols (β-sitosterol and an additional unknown sterol, 40% of total lipids) and diacylglycerol (43%) are the major lipid components in the PMVs of C. paradoxa. Minor components are PE, PC, PC and an unknown glycolipid. Fatty acid analysis revealed 12 different fatty acids, predominantly 16:0,18:0 and the polyunsaturated acids 20:4 and 20:5 (together 69% of total fatty acids), α-Linolenic acid, common in higher plant PMs, could only be detected as a minor constituent (< 1 %) of PMVs of C. paradoxa.     

Visualizing Bdellovibrio bacteriovorus by Using the tdTomato Fluorescent Protein

Bdellovibrio bacteriovorus is a Gram-negative bacterium that belongs to the delta subgroup of proteobacteria and is characterized by a predatory life cycle. In recent years, work has highlighted the potential use of this predator to control bacteria and biofilms. Traditionally, the reduction in prey cells was used to monitor predation dynamics. In this study, we introduced pMQ414, a plasmid that expresses the tdTomato fluorescent reporter protein, into a host-independent strain and a host-dependent strain of B. bacteriovorus 109J. The new construct was used to conveniently monitor predator proliferation in real time, in different growth conditions, in the presence of lytic enzymes, and on several prey bacteria, replicating previous studies that used plaque analysis to quantify B. bacteriovorus. The new fluorescent plasmid also enabled us to visualize the predator in liquid cultures, in the context of a biofilm, and in association with human epithelial cells.     

Effect of various organic compounds on synthesis of glucoamylase by an isolated strain Penicillium italicum

Yeast extract (0.5%) stimulates the production of glucoamylase and cell synthesis while methylene blue (0.1 mM) activates the synthesis of glucoamylase. Studies on the metabolic changes during fermentation of glucoamylase in a selected medium by P. italicum show that the rate of production of glucoamylase and cellular growth are greatly accelerated between 48 and 168 hr. Rapid growth of cells during this period may account for enhanced utilization of maltose and NH4NO3 from the medium. The acid production remains constant from 48 to 144 hr. Different forms of nitrogen decrease steadily. Although methylene blue stimulates the production of glucoamylase in the broth it has practically no effect on the rate of utilization of amino and total nitrogen from the broth.     

Kinetin regulates plant growth and biochemical changes during maturation and senescence of leaves,flowers, and pods of <Emphasis Type="Italic">Cajanus cajan</Emphasis> L.

Aspects of plant growth such as height, branch number, leaf number, leaf area, pod area, 100-seed mass, etc., were correlated with biochemical changes such as contents of chlorophyll (Chl), proteins, DNA, and RNA, and protease activity during development and senescent phases in leaves, flowers, and pods of Cajanus cajan L. cv. UPAS-120 after treatments with kinetin (Kn). A significant increase was noticed in branch number, leaf number, leaf area, and seed mass while other growth processes registered a small increase after Kn application. Effectiveness of 5 μM Kn was also noticed in minimizing the loss of Chls, proteins, and nucleic acids as well as reducing the protease activity during maturity and senescence. Chl a/b ratio maintained a high value up to 30-d followed by a decline in leaves while flowers registered much lower ratio at 20-d-age. Pods were unique in having relatively lower ratio of Chl a/b in comparison to leaves.     

Characterization and identification of chitinase producing Streptomyces venezuelae P10

In an attempt to isolate chitinase producers from soil, a streptomycete strain was found potent using natural chitin as the substrate. Chitinolytic activity was tested directly on agar plates, also with crude enzyme. Chitinase assay showed that the isolate could produce 0.8 U/ml of the enzyme. The morphological, cultural, physiological and biochemical characters of the isolate P10 were studied, and identified as Streptomyces venezuelae P10.     

Fluorescence-Based Comparative Evaluation of Bactericidal Potency and Food Application Potential of Anti-listerial Bacteriocin Produced by Lactic Acid Bacteria Isolated from Indigenous Samples

The aim of the present study was to ascertain the potency of anti-listerial bacteriocin produced by lactic acid bacteria (LAB) isolated from indigenous samples of dahi, dried fish, and salt-fermented cucumber. A total of 231 LAB isolates were obtained from the samples, of which 51 isolates displayed anti-listerial activity. The anti-listerial LAB were identified by PCR as Lactobacillus sp., Pediococcus sp., Enterococcus sp., and Lactococcus sp. PCR also enabled the detection of Class IIa bacteriocin-encoding genes such as enterocin A, pediocin, and plantaricin A in some of the LAB isolates. The culture filtrate from anti-listerial LAB isolates demonstrated bacteriocin-like inhibitory substance (BLIS) against common Gram-positive pathogenic bacteria such as Staphylococcus aureus, Enterococcus faecalis, and Bacillus cereus, and partial characterization of BLIS confirmed the production of bacteriocin by the LAB isolates. Sensitive fluorescence-based assays employing specific probes indicated the comparative potencies of the bacteriocin and clearly revealed the membrane-targeted anti-listerial activity of the purified bacteriocin produced by selected LAB isolates. The food application potential of plantaricin A produced by a native isolate Lactobacillus plantarum CRA52 was evidenced as the bacteriocin suppressed the growth of Listeria monocytogenes Scott A inoculated in paneer samples that were stored at 8?°C for 5?days.