Bioconversion of shrimp shell waste for the production of antioxidant and chitosan used as fruit juice clarifier

The pH, proteolytic activity, extent of demineralisation and deprotenisation of shrimp waste were studied during 7 days of fermentation using Pseudomonas aeruginosa A2. After 3 days, pH dropped from 7.0 to 4.4 and then remained constant. Simultaneously, a demineralisation of 92% was achieved. However, protease activity reached its highest level (1230 U mL^?1) after 1 day of incubation, and a protein removal of 90% was achieved. Chitin obtained was converted to chitosan. This chitosan, with 73% deacetylation, was tested for clarification of different fruit juices. It was observed that low concentrations of chitosan (below to 1%) greatly increase the clarity of juices without affecting the nutritional value. The antioxidant activity of the hydrolysates produced during fermentation was tested. Hydrolysate obtained after 3 days showed the strongest scavenging activity (90%), which was comparable to the positive control BHA; however, that obtained after 1 day exhibited the highest ferric‐reducing antioxidant power (OD 700 nm = 1.7).     

New alkaline trypsin from the intestine of Grey triggerfish (Balistes capriscus) with high activity at low temperature: Purification and characterisation

A highly alkaline trypsin from the intestine of Grey triggerfish (Balistes capriscus), with high activity at low temperature, was purified and characterised. The enzyme was purified to homogeneity using acetone precipitation, Sephadex G-100 gel filtration and Mono Q-Sepharose anion-exchange chromatography, with a 13.9-fold increase in specific activity and 41.3% recovery. The molecular weight of the purified alkaline trypsin was estimated to be 23.2 kDa by sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE) and size exclusion chromatography. Purified trypsin appeared as a single band on native–PAGE. Interestingly, the enzyme was highly active over a wide range of pH, from 9.0 to 11.5, with an optimum at pH 10.5, using Nα-benzoyl-DL-arginine-p-nitroanilide (BAPNA) as a substrate. The relative activities at pH 9.0, 11.5 and 12.0 were 86.5%, 92.6% and 52.4%, respectively. The enzyme was extremely stable in the pH range 7.0–12.0. In addition, the enzyme had high activity at low and moderate temperatures with an optimum at around 40 °C and had more than 80% of its maximum activity at 20 °C. The purified enzyme was strongly inhibited by soybean trypsin inhibitor (SBTI) and phenylmethylsulphonyl fluoride (PMSF), a serine protease inhibitor. The enzyme showed extreme stability towards oxidising agents, retaining about 87% and 80% of its initial activity after 1 h incubation at 40 °C in the presence of 1% sodium perborate and 1% H₂O₂, respectively. In addition, the enzyme showed excellent stability and compatibility with some commercial solid detergents.     

Trypsin from zebra blenny (Salaria basilisca) viscera: Purification,characterisation and potential application as a detergent additive

An alkaline trypsin was purified from the viscera of zebra blenny (Salaria basilisca) by ammonium sulphate (40?80% saturation) precipitation, Sephadex G-100, Mono Q-Sepharose and ultrafiltration. A yield of 12% with a purification-fold of 4.2 was obtained. The trypsin had an apparent molecular weight of 27 kDa. Soybean trypsin inhibitor and phenylmethylsulfonyl fluoride showed a strong inhibitory effect on the purified trypsin. Trypsin had maximal activity at pH 9.5 and 60 °C for the hydrolysis of Nα-benzoyl-dl-arginine-p-nitroanilide (BAPNA). It was stable at low temperatures and in the pH range of 7.0?12.0. The N-terminal amino acid sequence of the first 12 amino acids of the purified protease was IVGGRECTEPSQ. S. basilisca trypsin, which showed high homology with other fish trypsins, had a charged Arg residue at position 5, where Tyr is common in marine vertebrates and mammalian trypsins. The trypsin kinetic constants, K_m and k_cat for BAPNA, were 0.6 mM and 1.38 s^?1, respectively.     

Effect of Ramadan fasting on intragastric pH recorded during 24 hours in healthy subjects

OBJECTIVES: Lifestyle changes during Ramadan as the meals are taken exclusively in the evening, and nightly sleep is often delayed and shortened. The wake/sleep cycle is also modified. The aim of this study was to evaluate the influence of Ramadan on gastric acidity in healthy volunteers. METHODS: Nine healthy volunteers had 24-hour measurement of the gastric pH; 4 periods were compared: one week prior to Ramadan, day 10 and day 24 during Ramadan, and one month after Ramadan. The pH profiles and the [H+] activity (area under the curve) were measured during 24 hours, the night phase (5PM-8AM) and diurnal phase (8AM-5PM). RESULTS: The diurnal variations of the pH profile were more significant; the median pH was 2.3 prior to Ramadan, 1 at day 10 and day 24 and 1.6 one month after Ramadan. The 24-hours [H+] activity increased by 45% at day 10 of Ramadan compared with its level prior to Ramadan. This increase was mostly diurnal (+122%) and also nightly (+25%). The activity [H+] was steady during Ramadan. One month after Ramadan, the 24-hours [H+] activity was 23% higher than the one noted before Ramadan. CONCLUSIONS: This study shows that the conditions of feeding imposed by Ramadan are associated with an increase of the gastric acidity mainly in diurnal phase. These results do not explain the origin of the healthy volunteer digestive symptoms encountered during Ramadan.     

Spectral analysis for a degenerate tree and applications

The energy in network of strings subject to the feedback in a root is decreasing to zero if the network is non-degenerate and this decrease is never exponential. We study in this paper the case of degenerate network (i.e. the ratio of two lengths of strings is a rational number) with a feedback in the root of the network. Under this assumption, we determine the energy limit and we identify the best rate of decay with the spectral abscissa of the underlying semi-group generator. We identify the spectral abscissa as in terms of the largest modulus of the roots of a polynomial.     

Retention of cadmium ions from aqueous solutions by poly(ammonium acrylate) enhanced ultrafiltration

Cadmium removal from aqueous solution by polyelectrolyte enhanced ultrafiltration (PEUF) with relatively low transmembrane pressure was investigated at varying conditions of polyelectrolyte and cadmium concentrations, transmembrane pressure, ionic strength and pH. The poly(ammonium acrylate), with two average molecular weights (8000 and 15 000 Da) were used as polyelectrolyte. Flux declines during ultrafiltration of polyelectrolyte solutions. An effort has been made to evaluate these resistances independently at different operating conditions. The hydraulic membrane resistance is higher for processing solutions of PAmA₈ than solutions of PAmA₁₅. The study of ionic strength effect demonstrates that it decreases the retention of cadmium ions and increases the permeate flux. More than 99% of cadmium was retained for a NaNO₃ feed concentration less than 5 × 10⁻² mol L⁻¹. The pH effect study on the cadmium recovery revealed a maximum retention around 98% for pH 4.     

Chemical composition and characteristics of skin gelatin from grey triggerfish (Balistes capriscus)

Gelatin was extracted from the skin of grey triggerfish (Balistes capriscus) by the acid extraction process with a yield of 5.67 g/100 g skin sample on the basis of wet weight. The chemical composition and functional properties of gelatin were investigated. The gelatin had high protein (89.94 g/100 g) but low fat (0.28 g/100 g) contents. Differences in the amino acid composition between grey triggerfish skin gelatin (GSG) and halal bovine gelatin (HBG) were observed. GSG contained a lower number of imino acids (hydroxyproline and proline) (176 residues per 1000 residues) than HBG (219 residues per 1000 residues), whereas the content of serine was higher (40 versus 29 residues per 1000 residues, respectively). The gel strength of the GSG (168.3 g) was lower than that of HBG (259 g) (p < 0.05) possibly due to lower hydroxyproline content. Grey triggerfish skin gelatin exhibited a slightly lower emulsifying activity and water-holding capacity but greater emulsifying and foam stability, foam formation ability and fat-binding capacity than the halal bovine gelatin (p < 0.05). SDS-PAGE of GSG showed high band intensity for the major protein components, especially, α- and β-components and a similar molecular weight distribution to that of standard calf skin collagen type I.     

Distinct modifications by neurokinin1 (SR140333) and neurokinin2 (SR48968) tachykinin receptor antagonists of the N-methyl-D-aspartate-evoked release of acetylcholine in striosomes and matrix of the rat striatum

The effects of SR140333 and SR48968 (neurokinin1 and neurokinin2 tachykinin receptor antagonists, respectively) on the N-methyl-D-aspartate-evoked release of [3H]acetylcholine (previously formed from [3H]choline) were investigated in striosome-enriched areas and in the matrix of the rat striatum using an in vitro microsuperfusion method. In both striatal compartments, SR140333 and SR48968 did not modify the 50 microM N-methyl-D-aspartate-evoked release of [3H]acetylcholine. However, in low concentrations, both SR140333 (0.1 microM to 1 pM) and SR48968 (0.1 microM to 0.1 nM) markedly enhanced the 1 mM N-methyl-D-aspartate (+10 microM D-serine)-evoked release of [3H]acetylcholine in striosome-enriched areas. These responses were dopamine-dependent since they were not observed any more following the local blockade of D2 receptors by sulpiride or of dopamine synthesis by alpha-methyl-p-tyrosine. A dopamine-dependent disinhibitory effect (of lower amplitude) on the 1 mM N-methyl-D-aspartate (+10 microM D-serine)-evoked release of [3H]acetylcholine was also induced by SR48968 (0.1 microM to 0.1 nM) (but not by SR140333) in the matrix. In addition, in the matrix, as shown only in the presence of alpha-methyl-p-tyrosine, both SR140333 and SR48968 reduced the 1 mM N-methyl-D-aspartate (+10 microM D-serine)-evoked response and these non-dopamine-mediated inhibitory effects only occurred at the highest tested concentration (0.1 microM) of the antagonists. Indicating the specificity of these responses, the effects of SR140333 were reproduced by RP67580, another neurokinin1 receptor antagonist and, as expected from previous binding studies, corresponding SR140333 and SR48968 enantiomers were without effect. These results suggest that under potent stimulation of N-methyl-D-aspartate receptors, endogenously released substance P and neurokinin A (or related tachykinins) regulate differently the N-methyl-D-aspartate-evoked release of [3H]acetylcholine in striosomes and in the matrix. The inhibitory effects of these tachykinins on the evoked release of [3H]acetylcholine are mediated by dopamine. On the contrary, their facilitatory responses are only observed in the matrix under blockade of dopamine transmission.     

A novel aspartic protease from the viscera of Sardinelle (Sardinella aurita): Purification and characterisation

A novel aspartic protease was extracted from the defatted viscera of sardinelle (Sardinella aurita) and purified, with a 9.5-fold increase in specific activity and 23.3% recovery. The molecular weight of the purified enzyme was estimated to be 17 kDa by sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE). The purified enzyme appeared as a single band on native-PAGE. The optimum pH and temperature for protease activity were around 3.0 and 40 °C, respectively. The enzyme showed pH stability between 2.0 and 5.0 and retained more than 50% of its activity after heating for 30 min at 50 °C. The enzyme lost 90% of its activity after incubation with pepstatin A at room temperature, but was not inhibited by soybean trypsin inhibitor or phenylmethylsulfonyl fluoride. Its K_m value was determined to be 0.73 × 10⁻⁴ M using haemoglobin as a substrate. The N-terminal 12 amino acid sequence of the purified acidic protease was R V I I E D X D Q F C T. This sequence showed low homology with aspartic peptidases of several other species of fish, suggesting that the enzyme is a new aspartic protease.