Improving Bread Quality with the Application of a Newly Purified Thermostable α-Amylase from Rhizopus oryzae FSIS4.

A new thermostable α-amylase from Rhizopus oryzae FSIS4 was purified for first time and recovered in a single step using a three-phase partitioning (TPP) system. The fungal α-amylase, at a concentration of 1.936 U per kg of flour, was used in bread-making and compared to the commercial enzyme. The results showed a significant effect of the recovered α-amylase in the prepared bread and allowed us to improve the quality of the bread. The study indicated clearly that the recovered α-amylase is a potential candidate for future applications in the bread-making industry and in other food biotechnology applications.

Production and Properties of a Thermostable, pH-Stable Exo-Polygalacturonase Using Aureobasidium pullulans Isolated from Saharan Soil of Algeria Grown on Tomato Pomace.

Polygalacturonase is a valuable biocatalyst for several industrial applications. Production of polygalacturonase using the Aureobasidium pullulans stain isolated from Saharan soil of Algeria was investigated. Its capacity to produce polygalacturonase was assessed under submerged culture using tomato pomace as an abundant agro-industrial substrate. Optimization of the medium components, which enhance polygalacturonase activity of the strain Aureobasidium pullulans, was achieved with the aid of response surface methodology. The composition of the optimized medium was as follows: tomato pomace 40 g/L, lactose 1.84 g/L, CaCl20.09 g/L and pH 5.16. Practical validation of the optimum medium provided polygalacturonase activity of 22.05 U/mL, which was 5-fold higher than in unoptimized conditions. Batch cultivation in a 20 L bioreactor performed with the optimal nutrients and conditions resulted in a high polygalacturonase content (25.75 U/mL). The enzyme showed stability over a range of temperature (5-90 °C) with an optimum temperature of 60 °C with pH 5.0, exhibiting 100% residual activity after 1h at 60 °C. This enzyme was stable at a broad pH range (5.0-10). The enzyme proved to be an exo-polygalacturonase, releasing galacturonic acid by hydrolysis of polygalacturonic acid. Moreover, the exo-polygalacturonase was able to enhance the clarification of both apple and citrus juice. As a result, an economical polygalacturonase production process was defined and proposed using an industrial food by-product.

A novel killer protein from Pichia kluyveri isolated from an Algerian soil: purification and characterization of its in vitro activity against food and beverage spoilage yeasts.

A novel killer protein (Pkkp) secreted by a Pichia kluyveri strain isolated from an Algerian soil was active against food and beverage spoilage yeasts of the genera Dekkera, Kluyveromyces, Pichia, Saccharomyces, Torulaspora, Wickerhamomyces and Zygosaccharomyces. After purification by gel filtration chromatography Pkkp revealed an apparent molecular mass of 54 kDa with SDS-PAGE. Minimum inhibitory concentrations (MICs) of purified Pkkp exhibited a high in vitro activity against Dekkera bruxellensis (MICs from 64,000- to 256,000-fold lower than that exhibited by potassium metabisulphite) and Saccharomyces cerevisiae (MICs from 32,000- to 64,000- fold lower than potassium sorbate). No in vitro synergistic interactions (calculated by FIC index - Σ FIC) were observed when Pkkp was used in combination with potassium metabisulphite, potassium sorbate, or ethanol. Pkkp exhibited a dose-response effect against D. bruxellensis and S. cerevisiae in a low-alcoholic drink and fruit juice, respectively. The results of the present study suggest that Pkkp could be proposed as a novel food-grade compound useful for the control of food and beverage spoilage yeasts.

Pharmacological characterization of FE 201874, the first selective high affinity rat V1A vasopressin receptor agonist.

BACKGROUND AND PURPOSE: Distinct vasopressin receptors are involved in different physiological and behavioural functions. Presently, no selective agonist is available to specifically elucidate the functional roles of the V1A receptor in the rat, one of the most widely used animal models. FE 201874 is a new derivative of the human selective V1A receptor agonist F180. In this study, we performed a multi-approach pharmacological and functional characterization of FE 201874 to determine whether it is selective for V1A receptors. EXPERIMENTAL APPROACH: We modified an available human selective V1A receptor agonist (F180) and determined its pharmacological properties in cell lines expressing vasopressin/oxytocin receptors (affinity and coupling to second messenger cascades), in an ex vivo model (aorta ring contraction) and in vivo in rats (proliferation of adrenal cortex glomerulosa cells and lactation). KEY RESULTS: FE 201874 exhibited nanomolar affinity for the rat V1A receptor; it was highly selective towards the rat V1B and V2 vasopressin receptors and behaved as a full V1A agonist in all the pharmacological tests performed. FE 201874 bound to the oxytocin receptor, but with moderate affinity, and behaved as an oxytocin antagonist in vitro, but not in vivo. CONCLUSIONS AND IMPLICATIONS: On functional grounds, all the data demonstrate that FE 201874 is the first selective agonist of the rat V1A receptor isoform available. Hence, FE 201874 may have potential as a treatment for the vasodilator-induced hypotension occurring in conditions such as septic shock and could be the most suitable compound for discriminating between the behavioural effects of arginine vasopressin and oxytocin.

The effect of essential oil extracted from Nigella sativa (L.) seeds on human neutrophil functions.

This study was aimed to investigate the effect of essential oil on human neutrophil (HN) functions. The neutrophils were isolated on percoll gradients, counted, and tested for viability using the trypan blue exclusion method. The chemotactic response was based on a multiple blind well assay system. The control movement and chemotactic response of neutrophils to 0.1 microM fMLP were reduced at a dose-dependent manner. The essential oil significantly inhibited neutrophil chemotaxis from 0.05 to 0.5 mg mL(-1). The inhibitory concentrations (IC(50)) showing 50% inhibition to induced neutrophil chemotaxis, and control movement were 0.08 and 0.07 mg mL(-1), respectively. The human neutrophil elastase secretion was inhibited by essential oil at a concentration dependent manner from 0.5 to 2.5 mg mL(-1). The components of essential oil are potent inhibitors for polymorpho nuclear leukocytes functions. The observed inhibition of neutrophil functions occurred via intracellular pathway. Active serine protease could be essential for neutrophil responding process and/or signal transduction pathways.

Effects of essential oil extracted from Nigella sativa (L.) seeds and its main components on human neutrophil elastase activity.

The effects of essential oil extracted from Nigella sativa (L.) seeds and its main components on human neutrophil elastase (HNE) activity were investigated. Essential oil was extracted from N. sativa (L.) seeds using hydrodistillation. The yield was equal to 0.4%. Inhibition of HNE activity by essential oil was found to be dose dependent. The highest inhibitory concentration (HIC) of essential oil which caused total inhibition of HNE activity was 5.8 mg/ml. Microassays carried out to evaluate the inhibitory effect of major components of essential oil on HNE activity revealed that carvacrol (5-isopropyl-2-methylphenol) showed marked HNE inhibitory activity with a very low IC(50) value (12 microM). Based on these results, the inhibitory effects of essential oil on HNE activity are due to the presence of bioactive molecules, mainly carvacrol this compound is an inhibitor of HNE and could be considered as a natural antielastase agent and possible candidate for phytotherapy in the treatment of injuries that appear in some pathologic cases such as chronic obstructive pulmonary disease and emphysema.