Micellization of bovine beta-casein studied by isothermal titration microcalorimetry and cryogenic transmission electron microscopy     

Portnaya I  Cogan U  Livney YD  Ramon O  Shimoni K  Rosenberg M  Danino D 《Journal of agricultural and food chemistry》2006,54(15):5555-5561

The association behavior, critical micellization concentration (CMC), and enthalpy of demicellization (DeltaHdemic) of bovine beta-casein were studied, for the first time by isothermal titration calorimetry, in a pH 7.0 phosphate buffer with 0.1 ionic strength and in pure water. In the buffer solutions, the CMC decreased asymptotically from 0.15 to 0.006 mM as the temperature was raised from 16 to 45 degrees C. DeltaHdemic decreased with increasing temperature between 16 and 28 degrees C but increased from 28 to 45 degrees C. Thermodynamic analysis below 30 degrees C is consistent with the Kegeles shell model, which suggests a stepwise association process. At higher temperatures, this model exhibits limitations, and the micellization becomes much more cooperative. The CMC values in water, measured between 17 and 28 degrees C, decreased with increasing temperature and, expectedly, were higher than those found in the buffer solutions. beta-Casein micelles were visualized and characterized, for the first time in their hydrated state, using advanced digital-imaging cryogenic transmission electron microscopy. The images revealed small, oblate micelles, about approximately 13 nm in diameter. The micelles shape and dimensions remained nearly constant in the temperature range of 24-35 degrees C.  相似文献   

Identification and Characterization of a Novel Lectin from the Clam Glycymeris yessoensis and Its Functional Characterization under Microbial Stimulation and Environmental Stress     

Tatyana O. Mizgina  Irina V. Chikalovets  Valentina I. Molchanova  Rustam H. Ziganshin  Oleg V. Chernikov 《Marine drugs》2021,19(9)

Lectin from the bivalve Glycymeris yessoensis (GYL) was purified by affinity chromatography on porcine stomach mucin–Sepharose. GYL is a dimeric protein with a molecular mass of 36 kDa, as established by SDS-PAGE and MALDI-TOF analysis, consisting of 18 kDa subunits linked by a disulfide bridge. According to circular dichroism data, GYL is a β/α-protein with the predominance of β-structure. GYL preferentially agglutinates enzyme-treated rabbit erythrocytes and recognizes glycoproteins containing O-glycosidically linked glycans, such as porcine stomach mucin (PSM), fetuin, thyroglobulin, and ovalbumin. The amino acid sequences of five segments of GYL were acquired via mass spectrometry. The sequences have no homology with other known lectins. GYL is Ca²⁺-dependent and stable over a range above a pH of 8 and temperatures up to 20 °C for 30 min. GYL is a pattern recognition receptor, as it binds common pathogen-associated molecular patterns, such as peptidoglycan, LPS, β-1,3-glucan and mannan. GYL possesses a broad microbial-binding spectrum, including Gram-positive (Bacillus subtilis, Staphylococcus aureus) and Gram-negative bacteria (Escherichia coli, Vibrio proteolyticus), but not the fungus Candida albicans. Expression levels of GYL in the hemolymph were significantly upregulated after bacterial challenge by V. proteolyticus plus environmental stress (diesel fuel). Results indicate that GYL is probably a new member of the C-type lectin family, and may be involved in the immune response of G. yessoensis to bacterial attack.  相似文献   

Viability of Introducing Payments for the Collection of Wild Forest Mushrooms in Catalonia (North-East Spain)     

Irina Prokofieva  Elena Górriz-Mifsud  José-Antonio Bonet  Juan Martínez de Aragón 《Small-Scale Forestry》2017,16(2):147-167

Wild mushrooms are among the most valuable non-wood forest products in the world, and mushroom picking activities are well developed in many countries. Recent studies have demonstrated important links between forest management options and the productivity of mushrooms. Furthermore, there is evidence that the optimal forest management oriented at maximizing joint revenues from timber and mushrooms can lead to higher profits than the traditional timber-oriented management. The precondition for such management, however, is that forest owners derive benefits from mushrooms produced in their forests. In this paper, we discuss the feasibility of implementing payments for mushroom harvesting in Catalonia, North East Spain. As in many other European regions, mushroom picking in Catalonia is a long-standing tradition, which has been practiced with negligible limitations for centuries. The rising popularity of this activity in the last decades, however, has caused forest owners to voice concerns about the sustainability of this activity and about the impact it has on the private forest property. We document the results of a public opinion survey and contrast them with the issues emergent from forest owners’ interviews about the regulation of mushroom picking activities. Our results show that mushroom pickers support the idea of introducing payments for mushroom harvesting. Based on the survey and interview results, we also discuss the design aspects of payments for mushroom picking and articulate the key issues in their implementation.  相似文献   

6-Bromohypaphorine from Marine Nudibranch Mollusk Hermissenda crassicornis is an Agonist of Human α7 Nicotinic Acetylcholine Receptor     

Igor E. Kasheverov  Irina V. Shelukhina  Denis S. Kudryavtsev  Tatyana N. Makarieva  Ekaterina N. Spirova  Alla G. Guzii  Valentin A. Stonik  Victor I. Tsetlin 《Marine drugs》2015,13(3):1255-1266

6-Bromohypaphorine (6-BHP) has been isolated from the marine sponges Pachymatisma johnstoni, Aplysina sp., and the tunicate Aplidium conicum, but data on its biological activity were not available. For the nudibranch mollusk Hermissenda crassicornis no endogenous compounds were known, and here we describe the isolation of 6-BHP from this mollusk and its effects on different nicotinic acetylcholine receptors (nAChR). Two-electrode voltage-clamp experiments on the chimeric α7 nAChR (built of chicken α7 ligand-binding and glycine receptor transmembrane domains) or on rat α4β2 nAChR expressed in Xenopus oocytes revealed no action of 6-BHP. However, in radioligand analysis, 6-BHP competed with radioiodinated α-bungarotoxin for binding to human α7 nAChR expressed in GH₄C₁ cells (IC₅₀ 23 ± 1 μM), but showed no competition on muscle-type nAChR from Torpedo californica. In Ca²⁺-imaging experiments on the human α7 nAChR expressed in the Neuro2a cells, 6-BHP in the presence of PNU120596 behaved as an agonist (EC₅₀ ~80 μM). To the best of our knowledge, 6-BHP is the first low-molecular weight compound from marine source which is an agonist of the nAChR subtype. This may have physiological importance because H. crassicornis, with its simple and tractable nervous system, is a convenient model system for studying the learning and memory processes.  相似文献   

Inhibition of N-Type Calcium Channels by Fluorophenoxyanilide Derivatives     

Ellen C. Gleeson  Janease E. Graham  Sandro Spiller  Irina Vetter  Richard J. Lewis  Peter J. Duggan  Kellie L. Tuck 《Marine drugs》2015,13(4):2030-2045

A set of fluorophenoxyanilides, designed to be simplified analogues of previously reported ω-conotoxin GVIA mimetics, were prepared and tested for N-type calcium channel inhibition in a SH-SY5Y neuroblastoma FLIPR assay. N-type or Ca_v2.2 channel is a validated target for the treatment of refractory chronic pain. Despite being significantly less complex than the originally designed mimetics, up to a seven-fold improvement in activity was observed.  相似文献   

Bioprospecting Red Sea Coastal Ecosystems for Culturable Microorganisms and Their Antimicrobial Potential     

Soha Al-Amoudi  Magbubah Essack  Marta F. Sim?es  Salim Bougouffa  Irina Soloviev  John A. C. Archer  Feras F. Lafi  Vladimir B. Bajic 《Marine drugs》2016,14(9)

Microorganisms that inhabit unchartered unique soil such as in the highly saline and hot Red Sea lagoons on the Saudi Arabian coastline, represent untapped sources of potentially new bioactive compounds. In this study, a culture-dependent approach was applied to three types of sediments: mangrove mud (MN), microbial mat (MM), and barren soil (BS), collected from Rabigh harbor lagoon (RHL) and Al-Kharrar lagoon (AKL). The isolated bacteria were evaluated for their potential to produce bioactive compounds. The phylogenetic characterization of 251 bacterial isolates based on the 16S rRNA gene sequencing, supported their assignment to five different phyla: Proteobacteria, Firmicutes, Actinobacteria, Bacteroidetes, and Planctomycetes. Fifteen putative novel species were identified based on a 16S rRNA gene sequence similarity to other strain sequences in the NCBI database, being ≤98%. We demonstrate that 49 of the 251 isolates exhibit the potential to produce antimicrobial compounds. Additionally, at least one type of biosynthetic gene sequence, responsible for the synthesis of secondary metabolites, was recovered from 25 of the 49 isolates. Moreover, 10 of the isolates had a growth inhibition effect towards Staphylococcus aureus, Salmonella typhimurium and Pseudomonas syringae. We report the previously unknown antimicrobial activity of B. borstelensis, P. dendritiformis and M. salipaludis against all three indicator pathogens. Our study demonstrates the evidence of diverse cultured microbes associated with the Red Sea harbor/lagoon environments and their potential to produce antimicrobial compounds.  相似文献   

Diverse antimicrobial killing by Enterococcus faecium E 50-52 bacteriocin     

Svetoch EA  Eruslanov BV  Perelygin VV  Mitsevich EV  Mitsevich IP  Borzenkov VN  Levchuk VP  Svetoch OE  Kovalev YN  Stepanshin YG  Siragusa GR  Seal BS  Stern NJ 《Journal of agricultural and food chemistry》2008,56(6):1942-1948

An effective bacteriocin was identified and characterized. Lactic acid bacteria were screened against Campylobacter jejuni. One bacteriocin producer, Enterococcus faecium (NRRL B-30746), was studied. The isolate was grown, and the bacteriocin was purified to single-band homogeneity. Biochemical traits indicated that the peptide was a Class IIa bacteriocin, and it was named E 50-52. The bacteriocin had a molecular weight of 3339.7 and an isoelectric point of 8.0. The minimal inhibitory concentrations of E 50-52 against C. jejuni, Yersinia spp., Salmonella spp., Escherichia coli O157:H7, Shigella dysenteriae, Morganella morganii, Staphylococcus spp., and Listeria spp. ranged from 0.025 to 32 microg/mL. In therapeutic broiler trials, oral treatment with E 50-52 reduced both C. jejuni and Salmonella enteritidis by more than 100,000-fold in the ceca, and systemic S. enteritidis was reduced in the liver and spleen. The wide range of antibacterial activity of bacteriocin E 50-52 against pathogens provides a promising alternative to antibiotics.  相似文献   

Self-assembly of bovine beta-casein below the isoelectric pH     

Portnaya I  Ben-Shoshan E  Cogan U  Khalfin R  Fass D  Ramon O  Danino D 《Journal of agricultural and food chemistry》2008,56(6):2192-2198

Beta-casein is an intrinsically unstructured amphiphilic protein that self-assembles into micelles at neutral pH. This paper reports that beta-casein self-organizes into micelles also under acidic conditions. The protein association behavior and micelle characteristics at pH 2.6, well below the p I, are presented. The pH was found to strongly affect the micelle shape and dimensions. Cryogenic transmission electron microscopy (cryo-TEM) experiments revealed disk-like micelles of 20-25 nm in length and approximately 3.5 nm in height in acidic conditions. An aggregation number of 6 was determined by sedimentation equilibrium under these conditions. Isothermal titration calorimetry experiments verified the association below the p I and allowed determination of the micellization enthalpy, the critical micellar concentration, and the micellization relative cooperativity (MR). Small-angle X-ray scattering results at concentrations below the critical micellization concentration (CMC) suggest that the monomeric protein is likely in a premolten globule state at low pH. Calculations of the protein charge at acidic and neutral pH reveal a similar high net charge but considerable differences in the charge distribution along the protein backbone. Overall the results show that beta-casein is amphiphilic at low pH, but the distribution of charge along the protein chain creates packing constraints that affect the micelle organization, leading at concentrations above the CMC to the formation of disk micelles.  相似文献   

Rapid quantification of humic components in concentrated humate fertilizer solutions by FTIR spectroscopy     

Karpukhina  Evgeniya  Mikheev  Ivan  Perminova  Irina  Volkov  Dmitry  Proskurnin  Mikhail 《Journal of Soils and Sediments》2019,19(6):2729-2739

Journal of Soils and Sediments - The use of humic substances is under thorough discussion of state-of-the-art agricultural science. They are marketed mostly as concentrated aqueous solutions of...  相似文献   

Sex Specificity in Innate Immunity of Insect Larvae     

Irina Belousova  Sergey Pavlushin  Anna Subbotina  Natalya Rudneva  Vyacheslav Martemyanov 《Journal of insect science (Online)》2021,21(6)

The innate immunity of insects has been widely studied. Although the effect of sex on insect immunity has been extensively discussed, differences in immunity between the sexes of larvae insects remain largely unstudied. Studying larval sex differences in immunity may provide valuable information about the mechanisms underlying the insect immune system, which, in turn, can be valuable for the development and improvement of pest management. Here we compared the antibacterial activity in both the midgut tissue and cell-free hemolymph of Lymantria dispar L. (Lepidoptera: Erebidae) females and males at the larval stage without and after a challenge by entomopathogenic bacterium Bacillus thuringiensis Berliner. We also evaluated the sex-specific mortality of L. dispar induced by B. thuringiensis infection. We find that antibacterial activity in the midgut is activated by infection, but only in females. Thus, sex differences in immunity can have important effects even before sexual differentiation at adulthood.  相似文献