Effect of enterocin AS-48 in combination with high-intensity pulsed-electric field treatment against the spoilage bacterium Lactobacillus diolivorans in apple juice

Enterocin AS-48 was tested in apple juice against the cider-spoilage, exopolysaccharide-producing strain Lactobacillus diolivorans 29 in combination with high-intensity pulsed-electric field (HIPEF) treatment (35 kV/cm, 150 Hz, 4 μs and bipolar mode). A response surface methodology was applied to study the bactericidal effects of the combined treatment, with AS-48 concentration and HIPEF treatment time as process variables. At subinhibitory bacteriocin concentrations, microbial inactivation by the combined treatment increased as the bacteriocin concentration and the HIPEF treatment time increased (from 0.5 to 2.0 μg/ml and from 100 to 1000 μs, respectively). Highest inactivation (4.87 logs) was achieved by 1000 μs HIPEF treatment in combination with 2.0 μg/ml AS-48. While application of treatments separately did not protect juice from survivors during storage, survivors to the combined treatment were inactivated within the following 24 h of storage, and the treated samples remained free from detectable lactobacilli for at least 15 days at temperatures of 4 °C as well as 22 °C. The combined treatment could be useful for inactivation of exopolysaccharide-producing L. diolivorans in apple juice.     

Comparative effects of gamma and microwave irradiation on the quality of black pepper

Powdered black pepper from Egyptian markets, was irradiated with different recommended doses of gamma rays (5.0 and 10.0 kGy) and with microwaves for different periods (20, 40 and 75 s) to improve its hygienic quality. The most common bacterial isolates were of three generaBacillus, Clostridium andMicrococcus (7.5 × 10⁶), whereas the predominant fungi (7.8 × 10⁴) wereAspergillus species,A. glaucus, A. flavus, A. niger andA. ochraceus. Doses of gamma irradiation used (5.0 and 10 kGy) were sufficient to decrease spore-forming bacteria (SFB) and to inhibit the fungal flora and coliforms which contaminated the black pepper powder. Microwave treatments for 40 s and 75 s were of the same effectiveness whereas treatment for 20 s was less so. GLC analysis proved the presence of 31 peaks, only 19 compounds were identified as monoterpene hydrocarbons (56.21%), the major one being -phellandrene and limonene. Sesquiterpenes were also present, mainly -caryollphyllene (3.69%) as well as oxygenated compounds such as terpenol, geraniol, Me-chavicol, eugenol and anisol. Gamma irradiation at 5 kGy and 10 kGy respectively decreased the numbers of identified compounds from 21 (86.58% concentration) in untreated pepper to 16 (59.22% concentration), 15 (54.06% concentration). In comparison, microwave treatments, particularly for 40 s and 75 s, increased the concentration of the same compounds. The results obtained indicate that microwave treatment, under these conditions, is a safe and suitable technique for decontamination of black pepper which does not result in a great loss of flavour compounds, as compared with recommended doses of gamma irradiation.     

Isolation and identification of lipase producing thermophilic Geobacillus sp. SBS-4S: cloning and characterization of the lipase

A thermophilic microorganism, SBS-4S, was isolated from a hot spring located in Gilgit, Northern Areas of Pakistan. It was found to be an aerobic, gram-positive, rod-shaped, thermophilic bacterium that grew on various sugars, carboxylic acids and hydrocarbons at temperatures between 45°C and 75°C. Complete 16S rRNA gene sequence of the microorganism exhibited homology to various species of genus Geobacillus. A highest homology of 99.8% was found with Geobacillus kaustophilus. A partial (0.7 kbp) chaperonin gene sequence also showed a highest homology of 99.4% to that of G. kaustophilus whereas biochemical characteristics of the microorganism were similar to Geobacillus uzenensis. Based on biochemical characterization, 16S rRNA and chaperonin gene sequences, we identified SBS-4S as a strain of genus Geobacillus. Strain SBS-4S produced several extracellular enzymes including amylase, protease and lipase. The lipase encoding gene was cloned, expressed in Escherichia coli and the gene product was characterized. The recombinant lipase was optimally active at 60°C with stability at wide pH range (6-12). The enzyme activity was enhanced remarkably in the presence of Ca(+2). The K(m) and the V(max) for the hydrolysis of p-nitrophenyl acetate were 3.8mM and 2273 μmol min(-1)mg(-1), respectively. The ability of the recombinant enzyme to be stable at a wide pH range makes it a potential candidate for use in industry.     

Combined microwave and enzymatic treatments for ��-lactoglobulin and bovine whey proteins and their effect on the IgE immunoreactivity

Effect of combined microwave (MW) and enzymatic hydrolysis on the human immunoglobulin E (IgE)-binding properties of ??-lactoglobulin (??-lg) and other whey proteins (WP) was investigated. Separated ??-lg and full whey protein isolate (WPI) were hydrolyzed with trypsin, chymotrypsin, mixture of trypsin/chymotrypsin, and pepsin at three microwave power levels: 50?W during 1 and 5?min, 100 and 200?W during 1 and 3?min. The immunoreactivity of the obtained hydrolysates resulting from combined microwave protease treatment was assessed using sera of young patients allergic to bovine whey proteins. The application of microwave treatment at 200?W enhances the hydrolysis of ??-lg by pepsin in 3?min and decreases significantly its immunoreactivity. The extensive hydrolysis of the microwave-treated ??-lg and WPI with trypsin, chymotrypsin, and the mixture of trypsin with chymotrypsin did not have an impact on the IgE binding of the products obtained in all the studied conditions.     

Effects of postchill application of acidified sodium chlorite to control Campylobacter spp. and Escherichia coli on commercial broiler carcasses

Experiments were performed to assess the reduction of Campylobacter spp. and Escherichia coli in commercial broiler carcasses by postchill dip applications of acidified sodium chlorite. Carcass rinses were collected before the inside-outside-bird washer (IOBW), post-IOBW, postchill, and after the postchill application of acidified sodium chlorite. Prevalence and counts of Campylobacter spp. and E. coli were determined. The mean values for Campylobacter spp. and E. coli counts differed significantly at sampling sites. The IOBW reduced the bacterial counts significantly in only one experiment. The chiller reduced Campylobacter counts significantly in both experiments but failed to significantly reduce the counts of E. coli in one experiment. No major reduction in the prevalence after enrichment for Campylobacter spp. was detected post-IOBW or postchill. However, a significant reduction in Campylobacter spp. and in E. coli counts and Campylobacter spp. prevalence was seen after the postchill application of acidified sodium chlorite. These results demonstrate that the antimicrobial effect of acidified sodium chlorite applied postchill may be used to significantly reduce Campylobacter spp. and E. coli in commercial broiler carcasses. Postchill systems may eventually be used in different applications, such as mist, spray, or bath, which could be applied closer to the final stages in processing.     

Docking onto chromatin via the Saccharomyces cerevisiae Rad9 Tudor domain

An integrated cellular response to DNA damage is essential for the maintenance of genome integrity. Recently, post-translational modifications to histone proteins have been implicated in DNA damage responses involving the Rad9 family of checkpoint proteins. In budding yeast, methylation of histone H3 on lysine 79 (H3-K79me) has been shown to be required for efficient checkpoint signalling and Rad9 localization on chromatin. Here, we have used a rad9 Tudor mutant allele and cells mutated for Dot1, the H3-K79 methylase, to analyse the epistatic relationship between RAD9 and DOT1 genes regarding the DNA damage resistance and checkpoint activation pathways. Our results show that RAD9 is epistatic to DOT1 and suggest that it acts downstream of the Dot1 methylase in the damage resistance and checkpoint response. We have also found that the Tudor domain of Rad9 is necessary for in vitro binding to H3-K79me as well as Rad9 focal accumulation in response to DNA damage in vivo. In summary, our study demonstrates that the interaction between Rad9, via its Tudor domain, and methylated H3-K79 is required at two different steps of the DNA damage response, an early step corresponding to checkpoint activation, and a late step corresponding to DNA repair. The study further shows that the function of this interaction is cell cycle-regulated; the role in checkpoint activation is restricted to the G(1) phase and its role in DNA repair is restricted to G(2).     

Synthesis of α‐cellulose/polypyrrole composite for the removal of reactive red dye from aqueous solution: Kinetics and equilibrium modeling

In this work, a composite from α‐cellulose coated with conducting polypyrrole by in situ polymerization using potassium persulfate as oxidant was obtained. The composite was characterized by fourier transform infrared (FTIR) spectroscopy, cyclic voltammetry, UV/Vis spectroscopy, and scanning electron microscopy (SEM) analysis showed homogeneous coating of α‐cellulose with polypyrrole (PPy) to produce a composite with a conductivity of 3.5 × 10⁻⁵ S/m. Batch aqueous adsorption experiments of the reactive red 120 (RR120) dye onto the synthesized material were conducted. The results showed that this composite is an efficient adsorbent for RR120 dye removal. For the adsorption experiments set to an initial pH of 3.9, the adsorption capacity was 15.6 mg of dye/g of composite for an equilibrium concentration (in the liquid) of RR120 dye equal to 1,000 mg/L, whereas a value of 96.1 mg of dye/g of composite was obtained when the solution pH was set to 2.0 for the same equilibrium concentration. When performing adsorption experiments using pure α‐cellulose, dye adsorption was insignificant at any pH value. Adsorption isotherm for RR120 was described by a typical Freundlich model. The transient adsorption of RR120 on the synthesized composite was described by a general three‐resistance model that includes the transport on the film that surrounds the composite particles, diffusion inside the particles, and adsorption on the surface of the particles. A fitting of the uptake curves was performed allowing the estimation of values for the effective diffusivity, D₀, and the adsorption rate coefficient, k₁. For the adsorption experiments with an initial pH value set to 3.9, D₀ was estimated as 1.05 × 10⁻¹⁰ m²/s, whereas k₁ was 1.65 × 10⁻⁴ Lⁿ/g mg^{n − 1} s; the corresponding values of k₁ at pH = 2 and 9.0 were 3.18 × 10⁻⁴ and 5.16 × 10⁻⁵, respectively. POLYM. COMPOS., 36:312–321, 2015. © 2014 Society of Plastics Engineers     

Catalytic Sulfation of Betulin with Sulfamic Acid: Experiment and DFT Calculation

Betulin is an important triterpenoid substance isolated from birch bark, which, together with its sulfates, exhibits important bioactive properties. We report on a newly developed method of betulin sulfation with sulfamic acid in pyridine in the presence of an Amberlyst^®15 solid acid catalyst. It has been shown that this catalyst remains stable when being repeatedly (up to four cycles) used and ensures obtaining of sulfated betulin with a sulfur content of ~10%. The introduction of the sulfate group into the betulin molecule has been proven by Fourier-transform infrared, ultraviolet-visible, and nuclear magnetic resonance spectroscopy. The Fourier-transform infrared (FTIR) spectra contain absorption bands at 1249 and 835–841 cm⁻¹; in the UV spectra, the peak intensity decreases; and, in the nuclear magnetic resonance (NMR) spectra, of betulin disulfate, carbons С3 and С28 are completely shifted to the weak-field region (to 88.21 and 67.32 ppm, respectively) with respect to betulin. Using the potentiometric titration method, the product of acidity constants K₁ and K₂ of a solution of the betulin disulfate H⁺ form has been found to be 3.86 × 10^–6 ± 0.004. It has been demonstrated by the thermal analysis that betulin and the betulin disulfate sodium salt are stable at temperatures of up to 240 and 220 °C, respectively. The density functional theory method has been used to obtain data on the most stable conformations, molecular electrostatic potential, frontier molecular orbitals, and mulliken atomic charges of betulin and betulin disulfate and to calculate the spectral characteristics of initial and sulfated betulin, which agree well with the experimental data.