Purification and identification of novel antioxidant peptides from enzymatic hydrolysates of sardinelle (Sardinellaaurita) by-products proteins

In order to utilise sardinelle (Sardinellaaurita) protein by-products, which is normally discarded as industrial waste in the process of fish manufacturing, heads and viscera proteins were hydrolysed by different proteases to obtain antioxidative peptides. All hydrolysates showed different degrees of hydrolysis and varying degrees of antioxidant activities. Hydrolysate generated with crude enzyme extract from sardine (Sardinapilchardus) displayed high antioxidant activity, and the higher DPPH radical-scavenging activity (87 ± 2.1% at 2 mg/ml) was obtained with a degree of hydrolysis of 6%. This hydrolysate was fractionated by size exclusion chromatography on a Sephadex G-25 into eight major fractions (P₁–P₈). Fraction P₄, which exhibited the highest DPPH scavenging activity, was then fractionated by reversed-phase high performance liquid chromatography (RP-HPLC). Seven antioxidant peptides were isolated. The molecular masses and amino acids sequences of the purified peptides were determined using ESI-MS and ESI-MS/MS, respectively. Their structures were identified as Leu-His-Tyr, Leu-Ala-Arg-Leu, Gly-Gly-Glu, Gly-Ala-His, Gly-Ala-Trp-Ala, Pro-His-Tyr-Leu and Gly-Ala-Leu-Ala-Ala-His. The first peptide displayed the highest DPPH radical-scavenging activity (63 ± 1.57%; at 150 μg/ml) among these peptides.     

Angiotensin I-converting enzyme (ACE) inhibitory activities of sardinelle (Sardinella aurita) by-products protein hydrolysates obtained by treatment with microbial and visceral fish serine proteases

The angiotensin I-converting enzyme (ACE) inhibitory activities of protein hydrolysates prepared from heads and viscera of sardinelle (Sardinella aurita) by treatment with various proteases were investigated. Protein hydrolysates were obtained by treatment with Alcalase^®, chymotrypsin, crude enzyme preparations from Bacillus licheniformis NH1 and Aspergillus clavatus ES1, and crude enzyme extract from sardine (Sardina pilchardus) viscera. All hydrolysates exhibited inhibitory activity towards ACE. The alkaline protease extract from the viscera of sardine produced hydrolysate with the highest ACE inhibitory activity (63.2 ± 1.5% at 2 mg/ml). Further, the degrees of hydrolysis and the inhibitory activities of ACE increased with increasing proteolysis time. The protein hydrolysate generated with alkaline proteases from the viscera of sardine was then fractionated by size exclusion chromatography on a Sephadex G-25 into eight major fractions (P₁–P₈). Biological functions of all fractions were assayed, and P₄ was found to display a high ACE inhibitory activity. The IC₅₀ values for ACE inhibitory activities of sardinelle by-products protein hydrolysates and fraction P₄ were 1.2 ± 0.09 and 0.81 ± 0.013 mg/ml, respectively. Further, P₄ showed resistance to in vitro digestion by gastrointestinal proteases. The amino acid analysis by GC/MS showed that P₄ was rich in phenylalanine, arginine, glycine, leucine, methionine, histidine and tyrosine. The added-value of sardinelle by-products may be improved by enzymatic treatment with visceral serine proteases from sardine.     

Nonlinear dynamic analysis of curved beams

A computational procedure is presented for predicting the dynamic response of curved beams with geometric nonlinearities. A mixed formulation is used with the fundamental unknowns consisting of stress resultants, generalized displacements and velocity components. The governing semidiscrete finite element equations consist of a mixed system of algebraic and differential equations. The temporal integration of the differential equations is performed by using an explicit half-station central difference method. A procedure is outlined for lumping both the flexibilities and masses of the mixed model, thereby uncoupling all the equations of the system. The advantages of the proposed computational procedure over explicit methods used with the displacement formulation are discussed. The effectiveness and versatility of the proposed approach are demonstrated by means of numerical examples.     

Systems programming for data communications on minicomputers

This paper examines some of the special considerations which apply to the development of such software for minicomputers. These are treated under the following general categories:

• loading and throughput
• the external interfaces of the system software
• the internal structure of the system software
• developing the software

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Identification of BaO. A12O3.5TiO2 in the Ternary System BaO. A12O3-TiO2

A new identification and indexing for the phase BaAl₂Ti₅O₁₄ were accomplished using an X-ray diffraction technique. The new lattice parameters for the tetragonal lattice structure are: a₀=9.990 × 10^-10 m and c₀=12.264 × 10^-10 m, with a corresponding volume 1.224 × 10^-27 m³. The data provided by the Joint Committee on Powder Diffraction Standards are inconsistent both in lattice parameter values and Miller indices. The X-ray powder diffraction pattern of BaAl₂Ti₅O₁₄ was indexed using the LSUCR (least-squares unit cell refinement) computer program.     

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.     

Susceptibility of Date Fruits (Phoenix dactylifera) to Aflatoxin Production

Segments of fruits from 12 varieties of date (Phoenix dactylifera) and three stages of maturation were inoculated with a toxigenic strain of Aspergillus parasiticus. During growth at 28°C for 10 days, 8 varieties supported appreciable aflatoxin production at the Khalal stage—the most popular stage for direct human consumption, with a maximum value in excess of 300 μg g^-1 of fruit. Marked differences in susceptibility to infection and/or aflatoxin production were observed between varieties and/or stage of maturation. It was concluded that toxigenic aspergilli could proliferate on any date fruits that suffered mechanical damage in the field or during harvesting, and hence that such fruits should be considered as likely to be unfit for human or animal consumption. © 1997 SCI.     

Pressure death and tailing behavior of Listeria monocytogenes strains having different barotolerances

The objectives of this study were to investigate the variability among Listeria monocytogenes strains in response to high-pressure processing, identify the most resistant strain as a potential target of pressure processing, and compare the inactivation kinetics of pressure-resistant and pressure-sensitive strains under a wide range (350 to 800 MPa) of pressure treatments. The pressure resistance of Listeria innocua and nine strains of L. monocytogenes was compared at 400 or 500 MPa and 30 degrees C. Significant variability among strains was observed. The decrease in log CFU/ml during the pressure treatment was from 1.4 to 4.3 at 400 MPa and from 3.9 to >8 at 500 MPa. L. monocytogenes OSY-8578 exhibited the greatest pressure resistance, Scott A showed the greatest pressure sensitivity, and L. innocua had intermediate resistance. On the basis of these findings, L. monocytogenes OSY-8578 is a potential target strain for high-pressure processing efficacy studies. The death kinetics of L. monocytogenes Scott A and OSY-8578 were investigated at 350 and 800 MPa. Survivors at 350 MPa were enumerated by direct plating, and survivors at 800 MPa were enumerated by the most-probable-number technique. Both pressure-resistant and pressure-sensitive strains exhibited non-first-order death behavior, and excessive pressure treatment did not eliminate the tailing phenomenon.     

Purification, bacteriolytic activity, and specificity of beta-lytic protease from Lysobacter sp. IB-9374

Lysobacter sp. IB-9374, which was isolated from soil as a high lysyl endopeptidase-producing strain (Chohnanet al., FEMS Microbiol. Lett., 213, 13-20, 2002), was found to produce a beta-lytic protease capable of lysing gram-positive bacteria such as Staphylococcus aureus, Microccocuseus, and Bacillus subtilis. The Lysobacter strain secreted the beta-lytic protease into the culture medium at a 2.4-fold higher level than Achromobacter lyticus. The enzyme was highly purified through a series of six steps with a high yield. The enzyme was strongly inhibited by tetraethylene-pentamine and 1,10-phenanthroline. The purified enzyme lysed more efficiently almost all the gram-positive bacteria tested than lysozyme, lysostaphin, and mutanolysin. The enzyme was very similar to Achromobacter beta-lytic protease containing one zinc atom in terms of amino acid composition and N-terminal sequence. The nucleotide sequence revealed that the mature enzyme was composed of 179 amino acid residues with additional 198 amino acids at the amino-terminal end of the enzyme. The deduced amino acid sequence of the mature enzyme coincided with that of the Achromobacter enzyme, although the prepro-region showed a 41% sequence identity with the counterpart. These results indicate that Lysobacter sp. is a useful strain for an efficient large-scale preparation of beta-lytic protease capable of lysing bacteria.     

Simplex lattice mixture design approach on the rheological behavior of glucomannan based salep-honey drink mixtures: An optimization study based on the sensory properties

The rheological properties of salep drink sweetened with different honeys were measured using a controlled-stress rheometer. Mixture design experiments were used to study the effect of interactions among pine, flower and highland honeys on the rheological properties of salep-honey drink mixture (SHDM) samples. In addition, product optimization was carried out using the ridge analysis to determine the optimum mixture proportions based on sensory properties of SHDM samples. Flower honey was the component showing the highest effect on the consistency coefficient values of SHDM samples. The preference of panelists was more prominent for the SHDM samples including the higher concentrations of highland honey with respect to odour and overall preference parameters. Optimum values of pine, flower and highland honeys in the mixture were found to be 0–85%, 0–40% and 15–100%, respectively, with respect to sensory properties. In addition, ridge analysis results revealed that the SHDM should include 65% highland honey, 35% pine honey and no flower honey to obtain the maximum overall preference score (7.24). The consistency coefficient and flow behavior index values of the sample to get maximum overall preference score (7.24) were predicted to be 3.650 Pa sⁿ and 0.435, respectively.