In vitro digestibility of α-casozepine, a benzodiazepine-like peptide from bovine casein, and biological activity of its main proteolytic fragment

α-Casozepine is a peptide, corresponding to the sequence 91-100 of the bovine α(s1)-casein, displaying anxiolytic activity in the rat. The α(s1)-casein tryptic hydrolysate containing this peptide decreases stress effects after oral administration in various species including man. Therefore, the stability of this peptide toward gastric and pancreatic proteases has been assessed by using pepsin, chymotrypsin/trypsin, Corolase PP, pepsin followed by chymotrypsin/trypsin or pepsin followed by Corolase PP. α-Casozepine was slowly degraded by chymotrypsin, much more sensitive to pepsin and Corolase PP but not completely destroyed after 4 h kinetics. The bonds in the region 91 to 95 of the α-casozepine were totally resistant to hydrolysis by all studied proteases. Surprisingly, a fragment, corresponding to the sequence 91-97 and found in all the hydrolysis media in significant amount, possessed an anxiolytic activity in three behavioral tests measuring this parameter. This peptide could participate in the in vivo activity of α-casozepine.     

Effects of adding low levels of a disulfide reducing agent on the disulfide interactions of β-lactoglobulin and κ-casein in skim milk

Low concentrations of a disulfide reducing agent were added to unheated and heated (80 °C for 30 min) skim milk, with and without added whey protein. The reduction of the β-lactoglobulin and κ-casein disulfide bonds was monitored over time using electrophoresis. The distribution of the proteins between the colloidal and serum phases was also investigated. κ-Casein disulfide bonds were reduced in preference to those of β-lactoglobulin in both unheated and heated skim milk (with or without added whey protein). In addition, in heated skim milk, while the serum κ-casein was reduced more readily than the colloidal κ-casein, the distribution of κ-casein between the two phases was not affected.     

Broad antifungal activity of β-isoxazolinonyl-alanine,a non-protein amino acid from roots of pea (Pisum sativum L.) seedlings

Summary -(Isoxazolin-5-on-2-yl)-alanine (IA), a heterocyclic non-protein amino acid from root extracts and root exudates of pea seedlings, acts as a potent growth inhibitor of several eukaryotic organisms, including yeasts, phytopathogenic fungi, unicellular green algae, and higher plants. The antibiotic effect on baker's yeast was reversed by l-methionine, l-cysteine, and l-homocysteine. Phytopathogenic fungi such as Botrytis cinerea, Pythium ultimum, and Rhizoctonia solani grown on agar containing IA were inhibited in the growth of mycelia or in the production of sclerotia. In contrast, no significant inhibition of either Gram-positive or Gram-negative bacteria was observed. Rhizobium leguminosarum, the compatible microsymbiont of Pisum spp., and Rhizobium meliloti were able to tolerate up to 2.9 mM IA (500 ppm) without any effect on the growth rate. Bradyrhizobium japonicum even gave a positive chemotactic response to IA. The ecological significance of IA as a preformed plant protectant during the seedling stage of Pisum spp. and other IA-containing legumes is discussed.     

Effect of pH, NaCl, CaCl2 and temperature on self-assembly of β-lactoglobulin into nanofibrils: a central composite design study

The ability of certain globular proteins to self-assemble into amyloid-like fibrils in vitro opens opportunities for the development of new biomaterials with unique functional properties, like highly efficient gelation and viscosity enhancement. This work explored the individual and interacting effects of pH (1 to 3), NaCl (0-100 mM), CaCl(2) (0-80 mM) and heating temperature (80 to 120 °C) on the kinetics of β-lactoglobulin self-assembly and the morphology of resulting nanofibrils. Statistically significant (p < 0.05) interactions included CaCl(2)*temperature, NaCl*pH, CaCl(2)*pH, temperature*pH and NaCl*CaCl(2). Particularly notable was the very rapid self-assembly at pH 3 and the highly nonlinear effect of pH on self-assembly kinetics. Nanofibril morphologies ranged from long and semiflexible or curled and twisted to short and irregular. There did not seem to be a link between the kinetics of fibril formation and the morphology of fibrils, except at pH 3, where self-assembly was very rapid and fibrils were short and irregular, suggesting haphazard, uncontrolled self-assembly.     

Identification and molecular characterization of high-yielding,blast resistant lines derived from Oryza rufipogon Griff. in the background of ‘Samba Mahsuri’ rice

Genetic Resources and Crop Evolution - The wild species of rice, Oryza rufipogon, is a source of genetic variation for yield-enhancing traits and blast resistance. With the objective to genetically...     

Synthesis, analytical features, and biological relevance of 5-(3',4'-dihydroxyphenyl)-γ-valerolactone, a microbial metabolite derived from the catabolism of dietary flavan-3-ols

The physiological significance of 5-(3',4'-dihydroxyphenyl)-γ-valerolactone, an important metabolite derived from the catabolism of flavan-3-ols by gut microbiota, has been often overlooked due to the lack of the commercial standard. In the present work, this metabolite has been chemically synthesized, and its analytical parameters and antioxidant capacity have been determined in comparison to other chemical analogues [isomer 3-(3',4'-dihydroxyphenyl)-δ-valerolactone and γ-valerolactone] and other structurally related compounds [(+)-catechin, (-)-epicatechin, and 3-(3,4-dihydroxyphenyl)-propionic acid]. The synthesized compound was also used to perform a targeted analysis in samples collected during the in vitro fermentation of a grape seed flavan-3-ol extract with human fecal microbiota from three healthy volunteers. The time-course formation of 5-(3',4'-dihydroxyphenyl)-γ-valerolactone revealed large interindividual differences among volunteers, with concentrations ranging from 3.31 to 77.54 μM at 10 h of fermentation. These results are further discussed in view of the scarce reports quantifying 5-(3',4'-dihydroxyphenyl)-γ-valerolactone in in vitro fermentation studies, and pharmacokinetic and intervention studies.     

Purification and characterization of a novel β-1,3-1,4-glucanase (lichenase) from thermophilic Rhizomucor miehei with high specific activity and its gene sequence

Production, purification, and characterization of a novel β-1,3-1,4-glucanase (lichenase) from thermophilic Rhizomucor miehei CAU432 were investigated. High-level extracellular β-1,3-1,4-glucanase production of 6230 U/mL was obtained when oat flour (3%, w/v) was used as a carbon source at 50 °C. The crude enzyme was purified to homogeneity with a specific activity of 28818 U/mg. The molecular weight of purified enzyme was estimated to be 35.4 kDa and 33.7 kDa by SDS-PAGE and gel filtration, respectively. The optimal pH and temperature of the enzyme were pH 5.5 and 60 °C, respectively. The K(m) values of purified β-1,3-1,4-glucanase for barley β-glucan and lichenan were 2.0 mM and 1.4 mM, respectively. Furthermore, the gene (RmLic16A) encoding the β-1,3-1,4-glucanase was cloned and its deduced amino acid sequence showed the highest identity (50%) to characterized β-1,3-1,4-glucanase from Paecilomyces thermophila. The high-level production and biochemical properties of the enzyme enable its potential industrial applications.     

The effect of a mixture of two plant growth-promoting bacteria from Argentina on the yield of potato,and occurrence of primary potato diseases and pest – short communication

In 2017, we conducted a field experiment to test the influence of a mixture of two plant growth-promoting bacteria (Pseudomonas fluorescens and Azospirillum brasilense) on the yield of three potato varieties. In addition, the influence of the mixture on potato susceptibility to infection by the pseudofungus Phytophthora infestans and fungus Alternaria solani and to attacks by the Colorado potato beetle (Leptinotarsa decemlineata) was investigated. It was the first experiment of its kind in Europe. In the unusually hot and dry year, a positive influence on the yield (17%–31%) was found in all three varieties when the tubers were soaked in the bacterial mixture prior to sowing. We observed no significant differences in potato resilience/susceptibility to harmful organisms among the different treatments (soaked tubers, sprayed tubers, untreated tubers); however, there were significant differences in the productivity between the tuber varieties. The results of our research indicate the bacterial mixture used in this study has a high potential to support satisfactory potato yields under dry conditions and under low levels of infection by foliar fungal diseases and attacks by foliar insects.     

Distribution of Metals in Vadose Zone of the Alluvial Plain in a Mining Creek Inferred from Geochemical, Mineralogical and Geophysical Studies: The Beal Wadi Case (Cartagena�CLa Union Mining District, SE Spain)

The assessment of metals dispersal in polluted mining areas is a very complex issue, usually needing data from several analytical techniques in a joint approach. The present work focuses on the impact of the mining activity on lowlands alluvial plain sediments from an ephemeral creek, their role as source or sink of pollution and the spatial distribution of metals within the zone. In-depth distribution of elements (As, Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb, S, Ti and Zn) coming from mining activities was investigated by using X-ray fluorescence techniques and their mineralogical form using X-ray diffraction. A 2-D electrical resistance tomography field survey was carried out throughout at the creek bed to interpret the potential relationships between chemical, mineralogical and geophysical parameters. The application of leaching procedure (DIN 38414-S4 test) allows us to know the sediment??s heavy metals hazard and their potential mobility when changing redox conditions. From the results it was found that redox process of sulphur and the presence of carbonate influence the distribution of metals along the profile. In the present work, the precipitation of carbonates seems the most important process, especially for elements such as Zn and Mn. Secondary precipitation of sulphides enables the trapping of metals at sulphur-rich levels.