A soft thermo-reversible protein gel was studied with respect to failure. Flow curves recorded at constant shear-rates revealed a dynamic yield stress σy, seen as a stress plateau below about 10 s−1. When a shear stress below σy was applied to fractured gels, they reformed after a time that increased with increasing stress and diverged at σy. Application of shear stress to fresh gels led to an initial elastic response followed by creep. Following this creep regime, the strain stagnated below the dynamic yield stress σy, while for σ > σy failure was observed after a time that increased with decreasing stress and diverged at σ = σy. The time-to-failure dependence on the stress for σ > σy, with two distinct exponential scaling regimes, agrees with existing proposed theories for the fracture of colloidal strands.
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The identification of molecular structures of an arsenolipid is pivotal for its toxicological assessment and in understanding the arsenic cycling in the environment. However, the analysis of these compounds in a lipid matrix is an ongoing challenge. So far, only a few arsenolipids have been reported, including arsenic fatty acids (AsFAs) and arsenic hydrocarbons (AsHCs). By means of RP-HPLC-ICPMS/ESMS, we investigated Capelin oil (Mallotus villosus) for possible new species of arsenolipids. Twelve arsenolipids were identified in the fish oil including three AsFAs and seven AsHCs. Among the AsHCs, four that were identified had protonotated molecular masses of 305, 331, 347, and 359 and have not been reported before. In addition, the compounds with molecular formulas C20H44AsO+ and C24H44AsO+ were found in low concentrations and showed chromatographic properties and MS data consistent with cationic trimethylarsenio fatty alcohols. Derivatization by acetylation and thiolation coupled with accurate mass spectrometry was successfully used to establish the occurrence of this new class of arsenolipids as cationic trimethylarsenio fatty alcohols (TMAsFOH).
Sea cucumbers are used as healthy food, traditional medicine and dietary supplement. Sulphated polysaccharides (sPS) from their body wall possess variety of biological activities; however the immunomodulatory effects of sea cucumber polysaccharides remain unknown. Three sPS fractions were isolated from orange-footed sea cucumber (Cucumaria frondosa) and their effects on DC maturation investigated. DCs, matured in the presence of high molecular-weight FCF-1 (100 and 1000 µg/ml) with a monosaccharide composition of NANA, GlcNAc, Man, Gal, Fuc and GalNAc (40:13:12:12:9:8:3), secreted reduced levels of IL-10, IL-12p40 and IL-6 (only at 1000 µg/ml). Furthermore, allogeneic CD4+ T cells co-cultured with DCs matured with FCF-1 (100 µg/ml) secreted reduced levels of IFN-γ and increased levels of IL-17 than allogeneic CD4+ T cells co-cultured with DCs matured without FCF-1. These data suggest that FCF-1 can affect the DC maturation leading to increased Th17 and reduced Th1 activity; thus, increasing the Th17-mediated defence against yeast and extracellular bacteria.
The gene encoding the amylolytic enzyme Amo45, originating from a metagenomic project, was retrieved by a consensus primer-based approach for glycoside hydrolase (GH) family 57 enzymes. Family 57 contains mainly uncharacterized proteins similar to archaeal thermoactive amylopullulanases. For characterization of these family members soluble, active enzymes have to be produced in sufficient amounts. Heterologous expression of amo45 in E.coli resulted in low yields of protein, most of which was found in inclusion bodies. To improve protein production and to increase the amount of soluble protein, two different modifications of the gene were applied. The first was fusion to an N-terminal His-tag sequence which increased the yield of protein, but still resulted in high amounts of inclusion bodies. Co-expression with chaperones enhanced the amount of soluble protein 4-fold. An alternative modification was the attachment of a peptide consisting of the amino acid sequence of the mobile-loop of the co-chaperonin GroES of E.coli. This sequence improved the soluble protein production 5-fold compared to His6-Amo45 and additional expression of chaperones was unnecessary.
In this study, we present the discovery and characterization of a highly thermostable endolysin from bacteriophage Ph2119 infecting Thermus strain MAT2119 isolated from geothermal areas in Iceland. Nucleotide sequence analysis of the 16S rRNA gene affiliated the strain with the species Thermus scotoductus. Bioinformatics analysis has allowed identification in the genome of phage 2119 of an open reading frame (468 bp in length) coding for a 155-amino-acid basic protein with an Mr of 17,555. Ph2119 endolysin does not resemble any known thermophilic phage lytic enzymes. Instead, it has conserved amino acid residues (His30, Tyr58, His132, and Cys140) that form a Zn2+ binding site characteristic of T3 and T7 lysozymes, as well as eukaryotic peptidoglycan recognition proteins, which directly bind to, but also may destroy, bacterial peptidoglycan. The purified enzyme shows high lytic activity toward thermophiles, i.e., T. scotoductus (100%), Thermus thermophilus (100%), and Thermus flavus (99%), and also, to a lesser extent, toward mesophilic Gram-negative bacteria, i.e., Escherichia coli (34%), Serratia marcescens (28%), Pseudomonas fluorescens (13%), and Salmonella enterica serovar Panama (10%). The enzyme has shown no activity against a number of Gram-positive bacteria analyzed, with the exception of Deinococcus radiodurans (25%) and Bacillus cereus (15%). Ph2119 endolysin was found to be highly thermostable: it retains approximately 87% of its lytic activity after 6 h of incubation at 95°C. The optimum temperature range for the enzyme activity is 50°C to 78°C. The enzyme exhibits lytic activity in the pH range of 6 to 10 (maximum at pH 7.5 to 8.0) and is also active in the presence of up to 500 mM NaCl.
Nile tilapia (Oreochromis niloticus) farmed in recirculation aquaculture system (RAS) was filleted and packaged in 100% air and 50% CO2: 50% N2 modified atmosphere (MA) prior to storage at 1˚C and -1˚C for up to 27 days. Fillets were sampled regularly and analysed for headspace gas composition, sensory and microbial changes. Shelf life varied with apparent relation to storage temperature, package atmosphere and microflora. Pseudomonads were reported as the main spoilage organisms in tilapia fillets during chilled storage conditions. Sensory analysis of cooked samples as well as microbial growth indicated fillets packaged in 100% air had a shelf life of 13-15 days during storage at 1˚C and 20 days at -1˚C. At the end of shelf life in 100% air packaged groups, TVC and pseudomonads counts reached log 7 colony-forming units g-1 in flesh. Whereas in 50% CO2: 50% N2 packaged fillets, the lag phase and generation time of bacteria was extended and recorded counts of < log 4 colony-forming units g-1 up to 27 days of storage at both 1˚C and -1˚C. However, 50% CO2 : 50% N2 conditions restricted fillets shelf life to 23 days based on sensorial changes mainly fillets colour characteristics.
BACKGROUND
Although protein isolates have been proven as a potent raw material for protein hydrolysate preparation, the fishy odour associated with lipid oxidation is still detected. The remaining haemoglobin (Hb) in protein isolates can effectively induce lipid oxidation, leading to the formation of fishy odour in the resulting hydrolysate. The aim of this study was to elucidate the impact of Hb with different forms, oxyhaemoglobin (oxy-Hb) and methaemoglobin (met-Hb), on lipid oxidation and the development of fishy odour during hydrolysis of protein isolates.
RESULTS
During hydrolysis of protein isolate up to 120 min, non-haem iron content, peroxide value and thiobarbituric acid reactive substances slightly increased (P < 0.05). When oxy-Hb or met-Hb was incorporated, the marked increases in all parameters were observed, especially within the first 60 min of hydrolysis. The higher increases were obtained with the latter, suggesting that met-Hb was more pro-oxidative than oxy-Hb. However, no differences in degree of hydrolysis of all samples were observed (P > 0.05). The marked increases in the b*, ΔE*, ΔC* values, fishy odour/flavour and volatile compounds were also found in the resulting hydrolysate containing either oxy-Hb or met-Hb.
CONCLUSION
Hb, particularly met-Hb, induced lipid oxidation and the development of a fishy odour/flavour in fish protein hydrolysate. © 2013 Society of Chemical Industry
Chemical compositions and muddy compounds in dorsal and ventral muscles of Nile tilapia and broadhead catfish were comparatively studied. On a dry weight basis, Nile tilapia was rich in protein (93.1–93.8%), whilst broadhead catfish contained protein (55.2–59.5%) and lipid (36.6–42.4%) as the major constituents. Ventral portion had higher lipid or phospholipid contents with coincidentally higher geosmin and/or 2-methylisoborneol (2-MIB) contents. Geosmin was found in mince of Nile tilapia and broadhead catfish at levels of 1.5 and 3.2 μg/kg, respectively. Broadhead catfish mince had 2-MIB at level of 0.8 μg/kg, but no 2-MIB was detected in Nile tilapia counterpart. When pre-washing and alkaline solubilisation were applied for preparing protein isolate (PI), lipid and phospholipid contents were lowered with concomitant decrease in geosmin and 2-MIB contents. Protein hydrolysate produced from PI had a lighter colour and a lower amount of muddy compounds, compared with that prepared from mince. Therefore, PI from both Nile tilapia and broadhead catfish could serve as the promising proteinaceous material, yielding protein hydrolysate with the negligible muddy odour and flavour.
To reduce the practice of discarding commercially fished organisms, several measures such as a discard ban and extra allowances on top of landings quotas (“catch quota”) have been proposed by the European Commission. However, for their development and successful implementation, an understanding of discard patterns on a European scale is needed. In this study, we present an inter-national synthesis of discard data collected on board commercial, towed-gear equipped vessels operating under six different national flags spanning from the Baltic to the Mediterranean Seas mainly between 2003 and 2008. We considered discarded species of commercial value such as Atlantic cod (Gadus morhua), haddock (Melanogrammus aeglefinus), European hake (Merluccius merluccius), and European plaice (Pleuronectes platessa). Comparisons of discard per unit effort rates expressed as numbers per hour of fishing revealed that in the Mediterranean Sea minimum size-regulated species such as hake are generally discarded in much lower numbers than elsewhere. For most species examined, variability in discard rates across regions was greater than across fisheries, suggesting that a region-by-region approach to discard reduction would be more relevant. The high uncertainty in discard rate estimates suggests that current sampling regimes should be either expanded or complemented by other data sources, if they are to be used for setting catch quotas.
Contaminants have been analyzed in cod (Gadus morhua) since 1990 as part of the national monitoring program for the environmental conditions in the sea around Iceland. The aim of this study was to determine the temporal trends of persistent organic pollutants (POPs: polychlorinated biphenyls (PCBs), p,p′-dichlorodiphenyl dichloroethene (p,p′-DDE), hexachlorobenzene (HCB), hexachlorocyclohexanes (HCHs), chlordanes (CHLs) and toxaphenes (Tox)) and trace elements (As, Cd, Cu, Hg, Pb, Se and Zn) in cod over the last two decades at two different locations in the Arctic Ocean north of Iceland. The relationship between the contaminant concentrations and biological covariates was also determined. All of the POPs showed decreasing trends but the trace elements showed no clear signs of trend except arsenic which showed an increasing trend and zinc which showed a decreasing trend. The concentration of the POPs were lower or similar in the Icelandic cod compared to cod sampled in Norway, the Barents Sea and in the Baltic Sea, except for HCB which was higher in the Icelandic cod compared to the Norwegian cod. The concentration of the trace elements As, Cu, Hg and Zn were similar in the Icelandic cod compared to cod sampled in Norway and Greenland but the concentration of Cd was higher in the Icelandic cod. The inclusion of the biological covariates was found to be important for the statistical analysis. The POPs had a positive relationship with liver fat content but negative relationship with liver weight. The trace elements had a negative relationship with liver fat and liver weight except As which had positive relationship with liver weight. Only positive relationships were observed between the contaminant concentrations and length.